WO2004058046A2

WO2004058046A2 - Fluid treatment method, apparatus and system

Info

Publication number: WO2004058046A2
Application number: PCT/US2003/039973
Authority: WO
Inventors: Kenneth M. Johnson; Meir Dahan; Sohail Shadman; Mark B. Jones; Richard L. West; Ying-Cheng Lo; Tat Chi Mui; Alp Akonur; Atif Yardimci; Shawn Davis; Nikolay Zhizhin; Eric J. Kille; Tom Westberg; Judith Labonte; John F. Johnston; Julio Ottino; Abraham Leibson; David W. Pennington; Kenneth E. Pawlak; Julie A. Moriarity
Original assignee: Baxter International Inc.
Priority date: 2002-12-20
Filing date: 2003-12-16
Publication date: 2004-07-15
Also published as: AU2003300951A8; AU2003300951A1; WO2004058046A3

Abstract

Treatment apparatus, system and method are disclosed for treating a biological fluid, such as blood or blood components. The treatment may include but isnot limited to inactivation of pathogens in red cell concentrate. The system may include a disposable fluid circuit assembly and a reusable controller that controls flow through the fluid circuit for reconstituting, if necessary, a treating agent, combining the treating agent with a biological fluid and mixing the agent and biological fluid. An optional quenching agent may also be used.

Description

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FLUID TREATMENT METHOD, APPARATUS AND SYSTEM [0001] The present application hereby claims priority to and the benefit of earlier filed United States provisional patent application serial no. 60/435,146, filed December 20, 2002, and entitled "Static Mixing Apparatus and Method", and United States provisional patent application serial no. 60/448,217, filed February 19, 2003, and entitled "Red Cell Treatment", and United States provisional patent application serial no. 60/503,252, filed September 16, 2003, and entitled "Fluid Treatment Method, Apparatus and System," each of which is also hereby incorporated by reference .

[0002] The present invention generally relates to apparatus, methods and systems for the treating or processing of a biological fluid and, more specifically, to the ex vivo treating or processing of erythrocytes ("red blood cells") of human or other blood. Such treating or processing may be carried out for a variety of reasons including treating or processing the biological fluid to beneficially affect a function, quality, or characteristic thereof. More specific examples include treating or processing red cells for purposes of pathogen or white cell inactivation, prion removal/inactivation, universalizing the blood type of red cells, or other treating or processing of red cells or any combination of these. [0003] One preferred, but not exclusive, use of the present invention is treating or processing collected red cells to inactivate or destroy suspected pathogens or otherwise to render the pathogens inactive or ineffective to reduce the potential for disease transmission by transfusion of the red cells .

[0004] It is well known to collect red cells from healthy donors for subsequent transfusion to patients. For example, trauma patients often require transfusion of red cells in response to substantial blood loss. Transfusion of red cells may also be used to treat chronic anemia or blood loss due to surgery, including post-operative bleeding.

[0005] It is known, however, that blood components such as red cells, and other blood components, may harbor bacterial and/or viral pathogens that may be transmitted to the patient receiving the red cells. Blood borne pathogens may include, for example, hepatitis B virus, human immunodeficiency virus and a multitude of other bacterial and viral agents.

[0006] Although testing of donated blood components represents an important advance in the blood banking field, infectious agents can be transmitted from donors that have no detectable level of antibodies to the pathogen or nucleic acids of the pathogens. In addition, clerical errors and other mistakes may also expose patients to contaminated, incorrectly tested or mislabeled red blood cells. [0007] To address these shortcomings, significant efforts have recently been made in developing processes for inactivating pathogens in blood and blood components prior to transfusion. See, for instance, U.S. Patents Nos. 6,410,219; 6,171,777; 6,093,725; 5,559,250; and 5,691,132, which are hereby incorporated by reference. Baxter Healthcare Corporation of Deerfield, Illinois, has recently introduced a photoactivated system for inactivating pathogens in blood platelet concentrate. No pathogen inactivation process for red cells, however, has yet come to commercial fruition. [0008] Significant efforts have also been made in "universalizing" the blood type of red cells so they can be administered to any patient, regardless of blood type. Under the ABO typing system, type A blood normally cannot be administered to patients having blood type 0 or B. Similarly, type B blood normally cannot be administered to patients having blood type 0 or A. Type 0 blood, however, generally can be transferred to patients having type A, B or 0 blood. Unfortunately, according to Red Cross statistics, only about 38% of the United States population has type 0 blood. This means that much of the blood collected from the relatively small pool of healthy, voluntary blood donors can only be used in certain patients of a compatible blood type. Therefore, it would be highly desirable to have a treatment that reliably processes red cells to eliminate or reduce incompatibility.

[0009] Substantial work has been carried out to develop an enzymatic process to convert red blood cells of type A or B

(or AB) to the equivalent of an 0 type cell that could be transferred to any individual of type A, B or 0. In one such process, selected enzymes are added to collected red cells and incubated to remove the A and B related antigens that are found on the surface of the red cells. After removal, the antigens are washed away, leaving the equivalent of a type 0 red cell .

[00010] Also, certain techniques have been considered for prion removal or inactivation. Prions are the suspected agent in various spongiform encephalopathies such as Creutzfeld- Jacob Disease, Alphers Syndrome and other spongiform encephalopathies in both humans and animals. The prion removal/inactivation techniques include contacting the potentially prion-containing fluid with a sorption device having binding sites specific for binding prions. The fluid may be contacted with the sorption device by passing it through a porous sorption device or by storing the fluid in a vessel containing a sorption device in the form of a wafer, container wall surface or other configuration that allows contact between the fluid and the sorption device.

[00011] These are but a few of the various treatments or processes that may benefit from one or more features of the present invention, wherein at the end of the treatment a red cell (or other biological fluid) product is provided with a high level of confidence that the desired process steps or protocol has been successfully completed.

[00012] Accordingly, it is a general object of the present invention to provide methods, apparatus and systems useful in such treatment of biological fluids such as red cells and more particularly to provide methods, apparatus and system that can provide the needed high level of confidence that the desired treatment (such as pathogen inactivation or reduction, prion removal or blood type universaling) has been achieved. It is a more limited object to provide method, apparatus and system for inactivating pathogens in collected red cells.

[00013] Accordingly, although the objects, methods, apparatus and systems herein may be described in the context of specific applications, such as pathogen inactivation, it is understood that they also may have application in other treatment or processing of red blood cells or other biological fluids .

SUMMARY OF INVENTION [00014] The description of this invention is specifically in the context of red cell pathogen inactivation. However, it should be understood that certain aspects of this invention are not limited to pathogen inactivation in general or red cell pathogen inactivation in particular. Moreover, the present invention includes a number of different aspects which may have utility alone and/or in combination with other aspects. Accordingly, this summary is not exhaustive identification of each such aspect that is now or may hereafter be claimed, but represents an overview of the present invention to assist in understanding the present invention as set forth in the more detailed description. The scope of the invention is as set forth in the claims now or hereafter filed.

[00015] In accordance with one aspect of the present invention, a method for the ex vivo treatment of a biological fluid, such as red blood cells, is provided. In accordance with this method, a quantity of biological fluid, such as a specific collected unit or dose of red cells, a disposable fluid circuit assembly and a reusable controller are provided. The fluid circuit assembly and controller are cooperative to control fluid flow through the fluid circuit pursuant to a predetermined protocol, processor operation sequence including at least one treatment step to affect the red cells or other biological fluid.

[00016] In accordance with this method, the red cells, for example, are introduced into the disposable fluid circuit and processed through the fluid circuit assembly in accordance with the predetermined protocol, process or operation sequence, including treating the red cells in accordance with the treatment step to affect the red cells. As used herein, "affect" is intended to be broadly defined and to include any change or modification in a function, quality or characteristic of a biological fluid such as a quantity of red cells, including without limitation pathogen removal or inactivation, prion removal or inactivation, blood type universalizing or leukocyte removal or inactivation. [00017] In accordance with the present method, compliance with the process may be monitored to assure that the red cells or other biological components are processed through the fluid circuit in accordance with the predetermined protocol, process or operation sequence (hereinafter referred to as "predetermined sequence"). The monitoring system may include a data processing system with memory and a monitoring program stored in the memory. The monitoring program may have a listing of a plurality predetermined steps for treatment of a particular biological fluid in accordance with a desired treatment protocol or regimen. The system may further be in communication with treatment apparatus to receive communications or data regarding the predetermined steps. The data processing program may compare the information or data received to the requirements of the predetermined steps in the monitoring program to confirm that the biological fluid has been treated in conformity to the predetermined steps . [00018] In accordance with another aspect of the present invention, a disposable pre-assembled fluid circuit module is provided for use with a reusable control unit for treating red cells in accordance with a predetermined sequence. The disposable module may include a fluid inlet, a red cell (or other biological fluid) inlet, a fluid outlet, an interior chamber containing a red cell treating agent, an access member for accessing the interior chamber, a fluid pump and a flow path interconnecting the fluid inlet, red cell inlet, fluid outlet, pump and access member to provide fluid flow through the module in accordance with the predetermined sequence, under the control of the reusable control unit. [00019] In accordance with another aspect of the present invention, a reusable controller or control module is provided for controlling the processing of red cells or other biological component through a disposable fluid circuit module of the type summarized above. The reusable controller may include a receiving station for receiving the fluid circuit module into operative association with the controller module, a plurality of weigh stations, a pump actuator for actuating the pump on the fluid circuit module, a plurality of valves operable to control flow through the flow path in the fluid circuit module, an actuator for causing the sealed chamber to be accessed by the access member and a control system for automatically processing red cells or other biological component through the fluid circuit module in accordance with a predetermined sequence including at least one treatment step.

[00020] The pump on the fluid circuit module may take a variety of forms . In accordance with another aspect of the present invention, the pump may be syringe-type pump and comprises an elongated barrel defining a generally cylindrical bore that is open at one end. An elongated plunger extends through one end of the barrel, and an end of the plunger is disposed in and sealingly contacts the bore. The other end of the plunger is exterior to the bore for cooperation with the reusable control unit . The pump includes an elongated containment sleeve having opposed ends . The sleeve is disposed around the plunger, with one end of the sleeve being sealingly attached to the barrel and the other end of the sleeve being attached to the plunger to sealingly contain the one end of the plunger and to provide containment against a leakage from the open end of the barrel through which the plunger extends .

[00021] The fluid circuit module may also include one or more flow control members openable upon axial or bending movement to allow flow through the module or a portion thereof. For example, elongated frangible flow control members may be employed of the type having a weakened area between the opposite ends thereof, such that bending of the flow control member breaks the control member apart at the weakened area and opens the fluid flow path in which it is contained. In accordance with another aspect of the present invention, apparatus is provided for opening such a flow control member. Such apparatus, which may be employed in the reusable controller for automated opening of such a flow control member, may comprise an engagement member for acting on the frangible member in proximity to the weakened area and a linear actuator operably attached to the engagement member to move the engagement member linearly against the frangible member to fracture the member at the weakened area. Alternatively, such apparatus may comprise, for example, a rotor having at least 2 spaced members for receiving a frangible member therebetween and a rotary actuator adapted to rotate the rotor to bend at least one end of the frangible member relative to the weakened area to fracture the member. [00022] These and other aspects of the present invention are set forth in greater detail in the following detailed description of the following accompanying drawings. [00023] Figure 1 is a process flow chart depicting a process in accordance with one aspect of the present invention for pathogen inactivation .

[00024] Figure 2 is a disposable fluid circuit assembly that may be used to manually carry out a method of pathogen inactivation of red blood cells as described herein. [00025] Figure 3 illustrates a processing site layout that may be employed for treating red cells or other biological fluids in accordance with the present invention. [00026] Figure 4 illustrates another processing site layout that may be employed for treating red cells in accordance with the present invention.

[00027] Figure 5 illustrates insertion or assembly of a disposable fluid circuit module into a reusable controller for treating red cells or other biological fluids.

[00028] Figure 6 is a schematic flow diagram of the fluid circuit in the apparatus illustrated in Figure 5. [00029] Figure 7 is a plan view of the disposable fluid circuit module of Figure 5. [00030] Figure 8 is a perspective view of a rigid tubing organizer housing employed in the disposable fluid circuit module of Figure 5.

[00031] Figure 9 is an exploded perspective view of the tubing organizer housing of Figure 8, with one side of the housing separated from the other side of the housing and the fluid circuit components contained therebetween.

[00032] Figure 10 is a plan view of a tubing organizer and the associated fluid circuit components contained in the tubing organizer housing of Figure 8.

[00033] Figure 11 is a cross-sectional view, taken along line 11-11 of Figure 10, illustrating containment of the tubing organizer and associated fluid circuit within an overpouch or bag.

[00034] Figure 12 is an exploded perspective view of the tubing organizer and associated fluid circuit of Figure 10, with the overpouch removed.

[00035] Figure 13 is a plan view of the tubing organizer of Figure 12.

[00036] Figures 14 and 15 are side views illustrating the operation of tubing pinch valves employed in the illustrated apparatus for controlling flow through the fluid circuit. [00037] Figure 16 is side view of a piston-cylinder, syringe-type pump employed in the disposable fluid circuit module or assembly in Figure 5.

[00038] Figure 17 is an exploded perspective view of the syringe-type pump of Figure 16.

[00039] Figure 18 and 19 are cross-sectional views of a syringe-type pump that may be employed in the disposable fluid circuit assembly of Figure 5, showing the plunger or piston in inserted (Figure 18) and retracted (Figure 19) positions. [00040] Figure 20 is a side view of a vial container and container access assembly employed in the fluid circuit assembly of Figure 5.

[00041] Figure 21 is an exploded view of the vial container and access assembly of Figure 20.

[00042] Figure 22 and 23 are cross-sectional views of the vial container and access assembly of Figure 20 before and after accessing the vial contents.

[00043] Figure 24 is a perspective view of a static mixer that may be employed in the disposable fluid circuit assembly or module of Figure 5.

[00044] Figure 25 is a perspective view of an alternative static mixer that may be employed in the fluid circuit assembly of Figure 5. [00045] Figure 26 is a front view of the reusable controller or control module in Figure 5.

[00046] Figure 27 is a perspective view showing assembly of fluid flow tubing in a peristaltic pump located on the controller of Figure 26.

[00047] Figure 28 is a perspective view of the interior of the reusable controller or module, showing the inside surface of the front panel of the controller and slot housing, weigh stations and peristaltic pump associated with the front panel. [00048] Figure 29 is a perspective view of a slot housing and associated actuator assemblies employed in the controller of Figure 5.

[00049] Figures 30 and 31 are perspective views showing alternate positions of a linear-motion breaker apparatus that may be employed in the reusable controller of Figure 5 for opening a frangible flow control member in a fluid circuit assembly.

[00050] Figure 32 is side view of the vial actuator assembly (or subassembly) employed in the controller of Figure 5 and seen in Figures 28 and 29.

[00051] Figure 33 is a side view of the pump actuator assembly (or subassembly) employed in the controller of Figure 5 and seen in Figures 28 and 29. [00052] Figures 34 and 35 are perspective views of the valve actuator assembly (or subassembly) employed in the controller of Figure 5 and seen in Figures 28 and 29.

[00053] Figure 36 is an exploded perspective view of the valve actuator assembly of Figure 34.

[00054] Figure 37 is a perspective view of the apparatus of Figure 5, illustrating insertion of a disposable fluid circuit module into a receiving slot in a reusable controller. [00055] Figure 38 is a cross-sectional view of a weigh station employed in the reusable controller of Figure 5, with a container or pouch (part of the disposable fluid circuit module) depicted in the weigh station.

[00056] Figure 39 is a perspective view of another embodiment of a reusable mixing controller or control module. [00057] Figure 40 is a front view of the reusable controller or control module of Figure 39.

[00058] Figure 41 is a perspective view of the reusable controller or control module of Figure 39, showing a front door opened and a disposable fluid circuit assembly being inserted.

[00059] Figure 41A is a perspective view of the fluid circuit loading station of the controller shown in Figure 41. [00060] Figure 41B is a perspective view taken from the inside of the controller of Figure 41, showing the loading station housing and associated actuator assemblies.

[00061] Figure 41C is a perspective view of the valve actuator assembly employed in the controller of Figure 41.

[00062] Figure 41D is a perspective view of a pump actuator assembly employed in the controller of Figure 41.

[00063] Figure 42 is a perspective view of the reusable controller of Figure 41, with the disposable fluid circuit assembly fully inserted or loaded onto the face of the machine .

[00064] Figure 43 is a plan view of a disposable fluid circuit assembly of the type employed in connection with the reusable controller of Figure 39.

[00065] Figure 44 is a perspective view of a portion of the disposable fluid circuit assembly of Figure 43 which is inserted or loaded onto the face of the controller or control module of Figure 39.

[00066] Figure 45 is an exploded perspective view of the portion of the fluid circuit assembly shown in Figure 44.

[00067] Figure 46 is a perspective view of a tubing organizer on which a portion of the fluid flow circuit of Figure 44 is mounted. [00068] Figure 47 is a plan view of the fluid circuit and tubing organizer in an assembled condition, enclosed within a sealed overpouch or container, as shown in dashed lines. [00069] Figure 48 is a front view of the vial and vial access assembly employed in the disposable fluid circuit assembly of Figure 43.

[00070] Figure 49 is an exploded perspective view of the vial and vial access assembly shown in Figure 48. [00071] Figure 50 is a cross-sectional view of the vial and vial access assembly of Figure 48, showing one vial in a position prior to accessing the contents and the other vial in a position where the contents are being accessed. [00072] Figures 51 is a side view of a fluid flow manifold employed in the disposable fluid circuit assembly of Figure 43.

[00073] Figures 52-53 are perspective views of opposite sides of the manifold shown in Figure 51.

[00074] Figures 54 and 55 are cross-sectional views of the manifold shown in Figure 51, taken along lines A-A and B-B, respectively.

[00075] Figure 56 is a side view of a piston-cylinder or syringe type pump employed in the fluid circuit assembly of Figure 43. [00076] Figure 57 is an exploded view of the pump of Figure 56.

[00077] Figures 58-60 are cross-sectional views of the pump of Figure 56 showing the plunger in different positions.

[00078] Figure 61 is a perspective view of an alternative fluid circuit assembly portion that may be employed in the present invention.

[00079] Figure 62 is a perspective view of the fluid circuit assembly of Figure 61 with half of assembly removed for better view of the fluid circuit.

[00080] Figure 63 is a perspective view of a disposable slot housing and associated actuators that may be employed in a reusable controller for use with the fluid circuit assembly of Figure 61.

[00081] Figure 64 is plan view of a further alternative fluid processing circuit that embodies the present invention.

[00082] Figures 65-66 perspective views of static mixers that may be used in fluid circuit of Figure 64.

[00083] Figure 67 is a plan view of an alternative fluid processing circuit that embodies aspects of the present invention and employs a recirculation feature.

[00084] Figure 68 is a plan view of a still further fluid processing circuit that embodies aspects of the present invention and employs a recirculation feature. [00085] Figure 69 is yet another alternative fluid circuit assembly that embodies aspects of present invention.

[00086] Figure 70 is a perspective view of a portion of a fluid circuit assembly, showing an alternative mixing- arrangement embodying aspects of the present invention.

[00087] Figure 71 is an enlarged view of a portion of the fluid flow path in the circuit of Figure 70 for mixing in another direction.

[00088] Figures 72 is a further mixing device that may be employed for mixing a flow stream having a density gradient.

[00089] Figure 73 is a side view of a mixing member that may be employed to enhance mixing in different directions.

[00090] Figure 74 is an enlarged view of a static mixing section such as in Figure 70 showing added detail of apparatus for mixing in a direction in addition to radial.

[00091] Figure 75 is like Figure 74, illustrating a different embodiment of a static mixing section.

[00092] Figure 76 is an enlarged view of a further alternative static mixing section.

[00093] Figure 77 is a perspective view of alternative low- shear mixing apparatus that may be employed in treating red cells or other biological fluids.

[00094] Figure 78 is a partial top view of the apparatus of Figure 77. [00095] Figure 79 is a sequential perspective view showing an alternative low-shear mixing apparatus and method. [00096] Figures^' 80 and 81 are perspective views showing alternate positions of a linear-motion frangible breaker apparatus that may be employed in the reusable controller of Figure 5 for opening a frangible flow control member in a fluid circuit assembly.

[00097] Figure 82 is a perspective view of an alternate rotary-motion frangible breaker apparatus.

[00098] Figure 83-84 are cross-sectional views showing a frangible member in fluid tubing before and after breaking. [00099] Figure 85 is an exploded cross-sectional view of an alternative flow control device that may be employed in a disposable fluid circuit assembly of the present invention. [000100] Figure 86 is an assembled cross-sectional view of the flow control device of Figure 85 in a position blocking flow therethrough.

[000101] Figure 87 is a cross-sectional view of the flow control device of Figure 86, in a position allowing flow therethrough .

[000102] Figure 88 is an exploded cross-sectional view of a further alternative flow control device that may be employed in a disposable fluid circuit assembly of the present invention. [000103] Figure 89 is an assembled cross-sectional view of the flow control device of Figure 88, in a position blocking flow therethrough.

[000104] Figure 90 is cross-sectional view of the flow control device of Figure 89 in a position allowing flow therethrough.

[000105] Figure 91 is a cross-sectional perspective view of another flow control device that may be employed in the disposable fluid circuit assembly of the present inventions.

[000106] Figure 92 is a cross-sectional view of another alternative flow control device that may be employed in the disposable fluid circuit assembly of the present invention.

[000107] Figure 93 is an exploded perspective view of a further alternative flow control device that may be employed in the present invention.

[000108] Figure 94 is a cross-sectional view of an alternative vial holder and access assembly that may be employed in the present invention.

[000109] Figure 95 is a cross-sectional view of a further alternative vial holder and access assembly that may be employed in the present invention.

[000110] Figure 96 is a cross-sectional view of another alternative vial holder and access assembly that may be employed in the present invention. [000111] Figure 97 is an exploded perspective view of yet another vial holder and access assembly that may be employed in the present invention.

[000112] Figure 98 is a cross-sectional assembled view of the vial holder and access assembly of Figure 97.

[000113] Figure 99 is an exploded perspective view of still another alternative vial container and access assembly. [000114] Figure 100 is an assembled cross-sectional view of the vial container and access assembly of Figure 99. [000115] Figure 101 is a series of perspective views illustrating an incubation method and apparatus that may be employed in the present invention.

[000116] Figure 102 is a perspective view of alternative incubation and transfer method and apparatus that may be employed in the present invention.

[000117] Figure 103 is a perspective view of another alternative integrate incubation/transfer/adsorption apparatus .

[000118] Figure 104 is a perspective view of adsorption container carrier employed in the apparatus of Figure 102. [000119] Figure 105 is a perspective view illustrating the rotation or movement of the adsorption container carrier of Figure 104 through a limited arc to agitate the contents of the adsorption containers. [000120] Figure 106 is a schematic diagram of a data management system that may be employed to monitor and verify the biological fluid processing in accordance with process requirements or protocol .

DETAILED DESCRIPTION [000121] Turning now to a more detailed description of the present invention, the general or overall process embodying the present invention will be described first.

PROCESS OVERVIEW [000122] The present invention has application in a variety of red cell or other biological fluid treatment processes, but has particular application in pathogen inactivation in collected red cells, and is so described for purposes of illustration and not limitation. More specifically, Figure 1 depicts, in flow chart form, the various steps of a preferred process for inactivating suspected pathogens in collected concentrated human red cells. It is understood that this process may also be applicable to other treatment of red cells or other biological fluids or to inactivation of suspected pathogens in other blood components. The particular process shown in Figure 1 is the inactivation of suspected pathogens in a quantity (e.g., a unit) of collected red cells prior to transfusion to a patient.

[000123] In brief, the collected red cells are typically collected in concentrated form, in a manner well known in the blood banking field, with only a small amount of plasma remaining, in which they are suspended. The red cells are combined with a first solution, which may enhance storage of the red cells, at the time of collection or shortly thereafter.

[000124] The collected red cells, together with the first solution, are next combined with a pathogen inactivation agent and, optionally, a quenching agent . As illustrated, both of these components are reconstituted from a dry or powder form with a second or reconstitution solution. The pathogen inactivation agent could be another type of treating agent to affect the red cells (e.g. universalizing), if so desired. [000125] The concentrated red cells and first solution are then combined with the reconstituted pathogen inactivation agent and optional quenching agent. The combining of the reconstituted pathogen inactivation agent (and, optionally, the quenching agent) and the red blood cells is carried out in a manner to provide a high degree of reliability that all of the red blood cells have been properly combined with the pathogen inactivation agent. To further enhance the reliability of pathogen inactivation, a careful mixing of the red blood cells and the pathogen inactivation agent is employed which provides thorough mixing with minimum harm to the red blood cells. [000126] Following mixing of the red blood cells and pathogen inactivation agent, the combined mixture is stored or incubated for a period of time sufficient to allow the pathogen inactivation agent to inactivate any suspected pathogens harbored in the red blood cell suspension. The combined red blood cells and pathogen inactivation agent may be agitated during the period of incubation to further assure complete mixing, as well as to enhance the viability of the red blood cells and to prevent settling of the red cells in the suspension.

[000127] After incubation, the combined red cells and pathogen inactivation agent are optionally contacted with a sorption device to remove any unreacted inactivation agent, reaction by-products or any degradation products of the agent and/or any quenching agent. The sorption device may be based on absorption, adsorption or other sorption mechanism or principle which results in removal of the undesired components from the red blood cells. The red blood cell suspension may be left in contact with the sorption device for a period of hours, and to enhance contact with the sorption device and accelerate the process, the fluid may be agitated or circulated to better assure complete contact with the sorption material . [000128] After processing through the sorption step, the red blood cells may be transferred to a storage container for storage prior to subsequent transfusion to a patient . Alternatively, the sorption step may take place in a container which also serves as the storage container for the red cells. In any event, at the end of the process suspected pathogens have been inactivated or reduced and a very high degree of reliability is provided that the red cells administered to a patient will not carry or harbor pathogens that are capable of replication or capable of causing disease in the recipient.

[000129] The entire process of pathogen inactivation is preferably carefully monitored by a process control or data management system to assure that the resultant red cell product has undergone all of the steps of the pathogen inactivation process (or other treatment) , and that there has been rigorous adherence to a processing protocol to provide the desired level of reliability that suspected replicating pathogens in the red cells have been inactivated or reduced.

[000130] Turning now to a more detailed description of the process, each step will be explained in greater detail. RED CELL COLLECTION AND FIRST SOLUTION

[000131] It is anticipated that the collected red cells may be collected in a variety of ways, and may come from single or multiple donors. For example, one common way to collect red cells is referred to as a manual collection process. This is a process that many are familiar with and that is often used in local blood drives. In such a collection process, the donor donates whole blood, which is collected in a container, usually a flexible plastic bag or pouch. The collected whole blood, when returned to the laboratory or blood processing center, is typically processed by centrifugation to separate the various blood components. The higher density red cells are typically forced to the bottom of the pouch by the centrifugation, and the remaining plasma, platelets and white cells are expressed from the pouch for subsequent processing, leaving the collected concentrated red cells in the original pouch, where they are stored with a small amount of residual plasma. The manual collection process may typically be carried out using a multiple container system such as the Baxter Blood Pack product which has a preassembled series of containers with an attached phlebotomy needle for accessing a donor's vein, available from Baxter Healthcare Corporation of Deerfield, Illinois.

[000132] The red cells may also be collected employing what are commonly called automated or apheresis processes, wherein a donor's blood is circulated on a continuous or intermittent basis through a device that separates the red cells from the other blood components, saving the desired red cells, and either separately saving or returning the other blood components to the donor.

[000133] There are several automated systems available for red cell collection. The red cells may be collected in an automated process such with the Alyx , Amicus and/or CS-3000 Centrifugal Separators, available from Baxter Healthcare Corporation, Transfusion Therapies Division, Deerfield, Illinois 60015; the Spectra™ or Trima™ Apheresis System from Gambro Healthcare, in Lakewood, Colorado; the MCS+8150™ from Haemonetics Corporation of Braintree, Massachusetts; or the AS104 Cell Separator™ from Fresenius Hemocare, Inc. of Redmond, Washington.

[000134] Whether collected manually or automatically, it is contemplated that the red cells will be collected directly into the first storage solution or combined with the first solution shortly after collection. A combined, two part storage solution employed in the process shown in Figure 1 is described in detail in U.S. Patent No. 5,906,915, which is incorporated by reference herein. The two part solution has been sold as a combined solution by Baxter Healthcare Corporation of Deerfield, Illinois, under the trade name Erythro-sol™ or E-Sol™.

[000135] Other storage solutions may also be used in the red cell collection, and it is not required that it be a first part of a two part solution or other specialized solution. For example, the red cells may be initially collected in a storage solution that provides all the requirements for storage, such as Adsol^® solution available from Baxter Healthcare of Deerfield, Illinois, and does not require the addition of any other liquid to complement or complete the storage solution. In that event, another liquid such as water, saline, dextrose or other biologically compatible liquid, including an additional quantity of storage solution, may be used for reconstituting (including diluting) the treating agent and optional quenching agent.

[000136] A preferred first solution comprises sodium citrate, one or more buffers, adenine and mannitol. The first solution is substantially free of chloride and sugars. The buffers may include sodium phosphate monobasic and sodium phosphate dibasic. More specifically, the first solution may comprise 1 mMol/1 to about 2.2 mMol/1 adenine, approximately 20 mMol/1 to about 110 mMol/1 mannitol, approximately 2.2 mMol/1 to about 90 mMol/1 sodium citrate, approximately 1 mMol/1 to about 10 mMol/1 sodium phosphate/monobasic and approximately 5 mMol/1 to about 25 mMol/1 sodium phosphate dibasic.

[000137] The pH of the first solution is preferably above 7.0 and is physiologically compatible with the pH of the red blood cells. The pH of human red blood cells is approximately 7.4, and the pH of the first part solution part 1 may be approximately 7.4 ± 0.2 or thereabouts. As pointed out above, the first solution may be provided in the collection container into which red blood cells are collected directly from the donor, or may be combined with the red cells shortly after collection. Generally a unit of red cells comprises about 150-300 ml of concentrated red cells, more typically about 180-220 ml, and in accordance with this particular process, it is contemplated that a unit of red cells is combined either upon collection or shortly thereafter with about 80-100 ml, and more preferably about 94 ml, of the first storage solution.

[000138] Typically, after collection, the red cells are refrigerated to extend their shelf life. Prior to the next step in the inactivation process, it is desirable, although it may not be required, that the collected red cells (which are suspended in the first storage solution) be warmed to about room temperature. The red cells may be warmed with any commercially available warming device, or a custom system may be provided, depending on the particular needs of the user. The red cells are typically stored in a flexible plastic storage bag or pouch of polyvinyl chloride or other flexible plastic material, and may be warmed by placing the container on a warming surface of a heated platen or in a warm water bath. Commercially available platelet agitators, may be used to keep the red cells suspended as they warm. When warmed to room temperature or thereabouts, the solution of red cells, resuspended in the first storage solution, are ready for combining with the pathogen inactivation agent or other treating agent.

PATHOGEN INACTIVATION AGENT, QUENCHING AGENT AND SECOND SOLUTION

[000139] In the preferred embodiment, the pathogen inactivation agent, an optional quenching agent, and the second storage solution are combined before addition to the red cells.

PATHOGEN INACTIVATION AGENT

[000140] In the illustrated embodiment, the preferred red cell treating agent is a pathogen inactivation agent. More preferably, the pathogen inactivation agent is pH-activated anchor-1inker-effecter (ALE) or frangible-anchor-linker- effector (FRALE) compound, which irreversibly prevents replication of blood borne pathogens' RNA or DNA. The pathogen inactivation agent is a highly reactive acridine based compound, which penetrates the red cell membrane, pathogen membrane and/or coat and, through a reactive intermediate, cross links the nucleic acids of pathogens. The cross links inactivate the pathogens by preventing replication of their genomes. [000141] More specifically, the preferred pathogen inactivation agent is 3-alanine, N- (acridin-9-yl) , 2- [bis (2- chloroethyl) amino] ethyl ester. This agent and its use is described in more detail in U.S. Patents Nos. 6,093,725 and 6,410,219, which are incorporated by reference herein. [000142] The pathogen inactivation agent may be provided in a ready-to-use liquid form or may alternatively be provided in a form which requires reconstitution, including dilution, or other processing before addition to the red cells. For example, the pathogen inactivation may be in the form of a crystalline powder, a granulated powder, tablet, capsule, lyophilized powder, concentrated liquid or frozen liquid. The agent may be supplied in a wide variety of containers, such as bags, vials, rigid or flexible containers, syringe, or filled tubing or other appropriate container. In a preferred embodiment, about 10-100 mg, and more preferably about 50 mg of the agent in dry powder form is contained in a vial or other suitable container. Preferably, an excess amount of treating agent is provided to assure an adequate safety margin in the amount of treating agent to accommodate any variation in the amount of red cells.

[000143] Other pathogen inactivation agents may be used in the present invention, operating on similar or different principles, including but not limited to ethyleneimines monomers and polymers or similar compounds such as those disclosed in U.S. Patents Nos. 5,891,705; 6,093,564; 6,114,108; 6,150,109; 6,352,695; 6,559,321; 6,617,100; 6,617,101 and 6,617,157 and PCT application WO 00/02575, all of which are hereby incorporated by reference herein, and vitamin B related compounds such as riboflavin, alloxazine and isoalloxazine derivatives including but not limited to those disclosed in U.S. Patents Nos. 6,258,577; 6,268,120; 6,258,577; 6,277,337, all of which are also incorporated by reference herein.

QUENCHING AGENT [000144] The quenching agent in a preferred embodiment, which may be optional, is L-Glutathione, a naturally occurring tripeptide that does not penetrate the red cell membrane and pathogen membrane and/or coat . As with the pathogen inactivation agent, the L-Glutathione may be provided in various formulations and forms, including crystalline powder, liquid, low pH liquid, granulated powder, tablet, capsule, lyophilized powder or concentrated or frozen liquid and may come in the same variety of containers as the pathogen inactivation agent.

[000145] As set forth above, the purpose of a quenching agent is to react with any unused pathogen inactivation agent, inasmuch as there may be excess pathogen inactivation agent provided in order to better assure complete reaction treatment of the red cells. In the preferred embodiment as now contemplated, about 250-400 mg, and more preferably about 312 mg of L-Glutathione are provided in a vial or other suitable container.

[000146] Other quenching agents also may be used in the system, such as sodium thiosulfate and sodium thiophosphate, other thiosulfates and thiophosphates or compounds containing such moieties, cysteine, N-acetyl-cysteine, DTT, BHT, BHA, tyrosine, histidine, tryptophan, propyl gallate, and mercapto ropionyl glycine. Still other quenching methods that may used herein are disclosed in U.S. Patent No. 6,270,952, assigned to Cerus Corp. Other methods of employing quenching agents, other than adding a solution of the agent to the blood component, also may be employed. For example, a solid phase quenching system wherein the quenching agent is immobilized on a support structure may be employed in the system including, but not limited to, those quenching agents/structures disclosed in U.S. Patent No. 6,403,559, assigned to V.I, Technologies, Inc. The foregoing patents in this paragraph are hereby incorporated herein by reference .

SECOND SOLUTION [000147] The second solution, if required, is preferably aqueous-based and preferably includes a sugar selected from the group consisting of fructose and dextrose, to provide an energy source for long term storage of the red blood cells, after the inactivation or other treatment process is completed. For the above use, the second solution is preferably acidic and has a pH of approximately 5.8. [000148] As noted above, a preferred pathogen inactivation agent used in the illustrated embodiment is pH-activated. At an acidic pH, it is inactive or stable, and does not react excessively with the quenching agent. When the agent is added to the higher pH red cells (suspended in the higher pH first part of the storage solution) it becomes activated to carry out the inactivation process.

[000149] One of the additional benefits of having the second solution separate from the first solution relates to sterilization. For example, it is known that dextrose will degrade under the raised temperature of autoclaving unless the dextrose is maintained in an acidic medium. By providing storage solution in the form of two separate solutions, the second solution may be maintained at an acidic condition during heat sterilization and is not adversely affected (for purposes of sterilization) by the higher pH of the first solution. In other words, combining the first and second solutions before sterilization could result in adverse effects such as caramelization of the dextrose (i.e., glucose) during heat sterilization. This is avoided by maintaining the storage solution in two separate parts until after sterilization.

[000150] It is presently believed that about 10-50 ml, and preferably about 20-40 ml and possibly more preferably about

35-40 ml of the second solution will be sufficient to reconstitute the pathogen inactivation and quenching agents and provide sufficient volume for treatment of the likely volumes of red cells to be treated.

[000151] As noted earlier, the solution in which the red cells are collected may itself be a complete storage solution, and a separate second part of the solution may be unnecessary.

In that event, another biologically compatible liquid may be used to reconstitute the treating agent and/or quenching agent .

RECONSTITUTION OF PATHOGEN INACTIVATION AGENT AND QUENCHING AGENT

[000152] In accordance with the preferred embodiment of the illustrated process, the pathogen inactivation or other treating agent is provided in a powder form in a container or vial, as is the optional quenching agent, e.g., L-Glutathione.

To reconstitute the inactivation agent without prematurely activating it, the low pH second solution is used to reconstitute the inactivation agent. The reconstitution is preferably carried out by repeated circulating the second solution into and out of the vial (by repeated injection and withdrawal) until the inactivation agent is suspended or dissolved in the solution.

[000153] If the reconstitution is achieved by inserting a spike or other piercing member through a vial septum of an inverted vial, with the septum facing down, facing down reconstitution may be enhanced if the incoming stream of reconstitution liquid from the spike is directed at the "floor" of the vial (which is at the top when the vial is inverted) . The fluid stream into the vial may also have a relatively high velocity, such as a velocity greater than about 10 meters/second, with a Reynolds number in excess of 3000 and spike or piercing member upstream pressures of between about 20 and 60 psi, to further enhance reconstitution. Adequate reconstitution may be achieved with between about 3 and 4 ml of reconstitution liquid injected into the vial. The repeated injection and withdrawal of this liquid three or four times or more also serves to mix the reconstituted agent with the rest of the reconstitution (or second) liquid. The pumping action may also enhance mixing. [000154] The fluid exit ports in the spike or piercing member may be positioned so that they are just beyond the inside surface of the septum when the spike or piercing member is inserted. This location assists in creating a turbulent zone in the neck of the vial, where the bulk of the agent is located when the vial is inverted. The incoming reconstitution liquid initially provides a washing action against the inside surface of the walls of the vial. When located just inside the vial, the ports are also submerged in the reconstitution liquid, which aids in complete withdrawal of liquid and powdered agent. To further enhance reconstitution, a slight pull-back motion may be used after the spike or piercing member is inserted, so as to eliminate or reduce any internal depression, moat or trough that forms as the septum deflects slightly around the spike when it is inserted.

[000155] The spike or piercing member may employ a center point (as opposed to a needle cut) to reduce lateral forces and may have two or more exit ports around the central point to avoid areas of low turbulence. Reconstitution may be further enhanced if the spike or piercing member is inserted into the inverted vial at an acute angle to the vertical axis of the vial, such as a substantial acute angle, as an angle of about 30 degrees. Such an angle provides a turbulent, non- symmetrical flow pattern of reconstitution fluid within the vial, which is believed to enhance resuspension of the dry agent therein.

[000156] The inactivation agent and solution are then repeatedly injected into and withdrawn from the vial containing the quenching agent until the quenching agent is also resuspended in that solution. The features described above to enhance reconstitution of the treating agent may also be employed when reconstituting the quenching agent.

[000157] Other means or apparatus may be used for resuspending the inactivation agent and the quenching agent without departing from the present invention, as it is contemplated that a wide variety of apparatus could be used for this purpose. For example, the treating agent and quenching agent could be provided in a flexible pouch and reconstituted by adding liquid to each pouch and agitating the pouch manually or automatically to dissolve or dilute the treating agent and/or quenching agent.

[000158] Preferably, the second part of the two part solution comprises 1-15% dextrose solution, and more preferably an 8% dextrose solution, and when the inactivation agent and quenching agent are reconstituted, together comprise a volume of preferably about 20-40 ml and more preferably about 35-40 ml, which is sufficient to treat red cells in a range of anticipated collection volumes. As noted earlier, the volume of concentrated red cells may vary from about 150 to 300 ml, and there must be sufficient inactivation agent to accommodate that variation. [000159] When the pathogen inactivation agent and quenching agent are suitably reconstituted with the second solution, they are combined with the collected red cells suspended in the first solution.

COMBINATION OF INACTIVATION AGENT AND RED CELLS [000160] The reconstituted inactivation agent and optional quenching agent may be combined with the collected red cells in a wide variety of ways without departing from the present invention. The reconstituted inactivation agent and quenching agent (which together with the second solution comprise 10-50 ml of fluid, preferably about 20-40 ml and more preferably 35- 40 ml) may be added directly to the collected red cell product, for example by addition of the reconstituted agent directly into a container containing the collected red blood cells. Alternatively, the reconstituted inactivation agent and red cells may be conveyed separately into a mixing container where they are mixed together.

[000161] Preferably, however, the reconstituted inactivation agent and quenching agent are added to the red blood cells by a carefully controlled metering process. In such a process, the concentrated red blood cells are withdrawn from the container in which they have been stored at a relatively precisely controlled flow rate. The reconstituted inactivation agent and quenching agent are added to the red cell flow stream also at a relatively precisely controlled flow rate, so that the ratio of the flow rate of the reconstituted inactivation agent and quenching agent (with the second part of the solution) and the flow rate of the collected red blood cells is basically constant . An important benefit of metered combination of pathogen inactivation agent and red cells is that it provides a more predictable end point concentration of pathogen inactivation agent relative to red cells even when the volume of collected red cells varies. [000162] As was pointed out earlier, the collected concentrated red cells may have a volume from between 150-300 ml, although more typically the volume varies between 180-220 ml. By employing the metered introduction of inactivation agent into a metered flow rate of red cells, a constant ratio of inactivation agent to red cells is provided even when the amount of red cells varies from one donor to another. While effort is ongoing to determine the desired flow rate ratio, it is presently contemplated that the flow rates should be such as to result in a concentration of the most preferred inactivation agent from about 0.1-0.15 Mol to about 0.3 mMol, with the more desired concentration about 0.2 mMol. It is also contemplated that the concentration of L-Glutathione in the resultant product following mixing will be about 2.0 mMol. The precise concentration and control of L-Glutathione, however, as a quencher, is not believed to be as important as that of the inactivation agent. Of course, other concentration ranges for other pathogen inactivation agents and/or quenching agents may be appropriate and may be obtained by varying the amount of the agent, the volume of second solution used, and the relative flow ratio for mixing. Alternatively, dosing can be achieved by measuring the actual weights or volumes of red cells and reconstituted agents to be combined.

[000163] As presently contemplated, and as noted above, about 50 mg of pathogen inactivation agent and 312 mg of the quenching agent will be resuspended in approximately 35-40 ml of solution part 2, so as to provide sufficient volume of inactivation agent to accommodate varying volumes of collected concentrated red cells.

[000164] In one embodiment, the red blood cells are drawn from their source container by a peristaltic pump which provides a carefully metered flow rate of red blood cells. The reconstituted inactivation agent and quenching agent are drawn into a disposable syringe type pump (employing a piston- cylinder arrangement) which will precisely meter the flow of inactivation agent into the red blood cell stream. Although this is presently deemed a preferred approach, it is also understood that various means for combining the reconstituted inactivation agent and red blood cells could be used without departing from the present invention.

[000165] It should be noted that when employing the metering system described above, where a metered flow of pathogen inactivation agent is combined with a metered flow of red blood cells, it is preferred that the flow of inactivation agent begin first or first reach the junction, where the two streams are combined, to better assure that the desired concentration of inactivation agent is present in all the red cells and/or that all red blood cells are contacted by inactivation agent.

MIXING OF RED BLOOD CELLS AND INACTIVATION AGENT [000166] After the collected red blood cells and the reconstituted reactivation agent and quenching agent are combined, the combined fluids are subject to further mixing to provide a suitable level of confidence that complete mixing has taken place and that all the red blood cells have been treated by the inactivation agent.

[000167] As presently contemplated, in the arrangement where a metered flow of inactivation agent is added to a metered flow of red cells, the combined fluid stream is passed through a static mixing system which thoroughly mixes the combined fluid stream. By "static mixing," it is meant a mixing of the fluid stream without the need for moving parts such as impellers, blades or mechanically driven devices. [000168] Typically the static mixing occurs through the motive force of the flowing fluid stream itself. One such static mixing system is described in detail in U.S. Patent Applications Serial No.60/435, 146, entitled "Static Mixing Apparatus and Method," filed December 20, 2002, which is hereby incorporated by reference into this application. The static mixer shown in that application includes at least one and preferably a plurality of static mixing sections employing flow dividers that sequentially divide and rotate the combined fluid stream so as to mix the fluid stream, such as in a radial direction. A second mixing section may be provided, such as an accumulation chamber, which causes mixing of the combined fluid stream in a direction not limited to a radial direction, such as also in an axial direction, thereby better assuring a very high degree of homogeneity of the fluid stream.

[000169] The degree of mixing provided by the static mixer shown in the above-identified application is extremely high. For example, where the static mixer employs a series of mixing elements which sequentially stretch and fold the fluid stream, the mixing may be defined by the number 2ⁿ, where "N" represents the number of mixing elements. For example, with thirteen such mixing elements and fluid stream transformations, 2ⁿ is equal to 1.6 x 10⁴ and a hypothetical 1 cm fluid stream segment is repeatedly divided and subdivided so that it becomes less than 1 μm. It is further understood that when mixing pathogen inactivation agent and red cell fluid streams in this manner, mixing may be optimized by flowing the fluid stream in a laminar flow region or condition with a Reynold's number less than about 100.

[000170] Although the apparatus described in the above application represents the preferred mixing arrangement, it is also contemplated that other mixing techniques can be used to provide thorough mixing of the inactivation agent and the collected red cells. For example, the combined solutions may be mixed by expressing the solutions back and forth between two separate containers or bags, or between two or more compartments of the same container or bag. This can be done manually or can be done in an automated fashion by apparatus that alternately compresses and releases the containers or container portions to alternately express the solution back and forth between the containers or container compartments. [000171] Similarly, mechanical agitators into which a container of the combined red cells and inactivation agent are placed may also be employed to provide thorough mixing of the solution. For purposes of the present invention, however, the means by which the collected red cells and pathogen inactivation are mixed is not limited to one particular means or apparatus. The objective is to obtain a suitable concentration of pathogen inactivation or other treating agent in the collected red cells or other biological fluid and to provide sufficient mixing, whatever apparatus if any is used, to provide the needed high degree of assurance that the inactivation agent has been suitably mixed through-out the biological fluid.

INCUBATION

[000172] After combination of the pathogen inactivation agent and the collected red cells and mixing of those solutions, the combined collected red cells, pathogen inactivation agent, quenching agent, and storage solution (now first and second solutions combined) are incubated for a period of time sufficient to assure that the inactivation process has taken place. Presently, it is contemplated that a container containing the combined solution will be incubated in an environmentally controlled area having an ambient temperature of between about 19-25° C, for about 1-24 hours and more preferably 12 hours, although more or less time may be required or sufficient.

[000173] To prevent the red cells from accumulating in one part of the container and thereby diminishing the reliability of the inactivation process, the combined solution is preferably agitated during the incubation period. A variety of apparatus may be employed to agitate the container during the incubation period. For example, the container may be placed on a commercially available platelet incubator/agitator which oscillates back and forth, keeping the contents of the container in an agitated state and the red cells generally suspended throughout the solution. Alternatively, custom systems may be designed that repeatedly turn the container or invert it throughout the incubation period.

SORPTION

[000174] After incubation, the combined red cell and inactivation solution is optionally treated to remove any unused pathogen inactivation agent or quenching agent and any reaction or degradation product of the agent. More specifically, in the illustrated embodiment, the combined solution is contacted with a sorption device, which may operate by adsorption, absorbtion or other sorption mechanisms to cleave to or otherwise remove any remaining inactivation agent and degradation or reaction by-products.

[000175] There are several different ways the combined solutions may be contacted with a sorption device. For example, the sorption device may be in the form of a filter through which the solution is passed, either on a one time basis or repeatedly to carry out the sorption process. In a more preferred embodiment, the sorption is carried out by a compound adsorption device ("CAD") in the form of a wafer or insert that is located in a container, into which the treated red cells are transferred.

[000176] One such compound adsorption device is described in detail in U.S. Patent Application Serial No. 60/364,289 filed March 14, 2002, entitled "Compound Removal Device," which is hereby incorporated by reference into this application. The combined red cells and pathogen inactivation agent are stored in contact with the adsorption device for a sufficient period of time to complete the adsorption process. In the illustrated method, the treated red cells are stored in a container with the adsorption device for approximately 1-24 hours, more preferably about 8 hours at a temperature ranging from about 19-25° C.

[000177] To accelerate the adsorption process and to increase the adsorption kinetics, the container of treated red cells and adsorption device may be continually agitated during the storage period to enhance mixing and contact of the solution with the adsorption device. Various apparatus or techniques may be used to agitate the solution during the adsorption process. These apparatus may include an orbital shaker, a platelet agitator, rotary agitator or a device such as a Wheaton roller for continuously turning the container (available from Wheaton Science Products of Millville, New Jersey) . Alternatively, the container may be manually or automatically massaged or squeezed, the solution may be transferred back and forth between separate containers or separate compartments in the same container.

[000178] Further apparatus may be provided to automatically open a fluid passage between the incubation container and the adsorption container and transfer the red cells, such as by inverting the containers to allow gravity assisted flow from one container to another.

[000179] After the sorption process is complete, the red cells may be transferred to another container for storage until transfusion or, alternatively, may remain in the same container in which the sorption process has taken place. At the end of the process, however, the red cells have undergone a pathogen inactivation treatment which provides assurance to an extent not previously available that any suspected pathogens contained or harbored in the red cells have been inactivated and present no significant risk to a patient receiving a transfusion of the red cells.

PROCESS CONTROL

[000180] The method of red cell treatment described above or other biological fluid treatment is preferably controlled and/or monitored by an electronic data management system that monitors and/or controls each stage of the process to best assure that when the process is complete that it has been performed in accordance with rigorous adherence to the required processing steps to achieve the desired pathogen inactivation. The integrated data management system may include, for example, bar code tracing of each quantity or unit of red cells throughout the process, monitoring by appropriate sensors or detectors in the apparatus employed in carrying out the process to confirm processing in accordance with the predetermined steps of a stored protocol in the data system memory, and confirmation by pre- and post-processing assays. Such a system also permits the archival of data for later access, as needed, for review of the processing carried out on any individual quantity or unit of red cells that have been treated or for statistical analysis or other uses.

MANUAL FLUID CIRCUIT ASSEMBLY [000181] The process described above may be carried out using various apparatus and systems ranging from entirely manual to highly automated. Figure 2 illustrates a fluid circuit assembly 50 that may be used in manually carrying out a biological fluid treatment procedure and, in particular, a red cell pathogen inactivation procedure as described above. The fluid circuit assembly 50 is preferably an integral, closed system, pre-assembled, pre-sterilized and ready for connection to a quantity (such as a collected unit) of concentrated red cells . [000182] The illustrated fluid circuit assembly 10 includes a series of flexible plastic containers or pouches, connected by tubing, including a reconstitution liquid container 12, an optional quenching agent container 14, a pathogen inactivation agent container 16, a mixing container 18, an incubation or reaction container 20, an adsorption container 22, and a storage or transfusion container 24. The container 12 of reconstitution liquid may contain a selected quantity of dextrose, which may be the second part of a two part red cell storage solution as discussed above. The container 12 is connected in series to the quenching agent container 14 and the pathogen inactivation agent container 16, which may contain, respectively, a quenching agent, specifically glutathione, and a pathogen inactivation agent, i.e., an ALE compound, both in powder (or other) form, as described above.

[000183] To begin the reconstitution, a standard frangible flow control member (or other flow control device) in the connecting tubing is opened and the dextrose from container 12 is expressed into the quenching agent container 14. The quenching agent is reconstituted by agitating and/or shaking the container 14. After the quenching agent is reconstituted, a flow control member downstream of the quenching agent container is opened and the combined quenching agent and dextrose are expressed into the pathogen activation agent container 16, where the liquid is agitated in the container by shaking or squeezing or otherwise manipulating or massaging the container to reconstitute the pathogen inactivation agent. [000184] Next, the reconstituted inactivation agent and quenching agent are mixed with a collected quantity of red cells. The pathogen inactivation agent container 16 is connected by tubing to the mixing container 18. A junction 26 in the tubing includes a branch tubing segment adapted for sterile connection to a container or pouch 28 containing a quantity of collected red cells. The reconstituted pathogen inactivation agent (and optional quenching agent) and red cells are drained or expressed from their respective containers into the mixing container 18. After transfer into mixing container is complete, the tubing into the mixing container may be sealed and severed from the upstream containers and tubing.

[000185] The mixing container 18 is then gently agitated by repeatedly squeezing or twisting or by gently shaking or massaging the container. Preferably the agitation continues for at least several minutes with a hundred or more agitations to assure generally complete mixing. After the mixing is complete, a flow control member is opened and the combined red cells and pathogen inactivation agent are expressed into the incubation or reaction container 20. [000186] To allow the pathogen inactivation agent sufficient time to treat the red cells, the container 20 of combined fluids is incubated or stored at the desired temperature for up to 12 hours or more. Preferably the container is gently agitated during incubation to enhance the reaction and to keep the red cells suspended. After the incubation is complete, the combined red cells and inactivation agent are drained into the adsorption container 22.

[000187] The adsorption container has a pre-inserted adsorption media, such as a wafer or the like, as described earlier. The red cells are stored in the adsorption container for a time sufficient to allow adsorption of any unused pathogen inactivation agent, quenching agent or reaction byproducts. It is anticipated that about 8 hours storage in the adsorption container, with gentle agitation, is sufficient to complete the adsorption process. At that point, the treated red cells may be transferred into the transfusion container 24 for refrigerated storage until required for transfusion to a patient .

PROCESSING CELL

[000188] Figures 3 and 4 depict possible processing or production cells or production areas in which a red cell or other biological fluid treatment as described above may be carried out in a more efficient and automated manner using a more preferred system and apparatus of the present invention. The production cell or area 30 may comprise four functional stages or stations - a receiving/preparing station 32, a mixing station 34, an incubation/adsorption station 36 and a final processing/checking station 38. Although a variety of station arrangements may be used, Figure 3 illustrates a compact and efficient layout, wherein the receiving, incubating and final check stations are arrayed sequentially and located generally around the mixing station 34. [000189] Typically, for example, a quantity of red cells, such as a single collected unit or container, arrives first at the receiving/preparing station 32. There, the attendant may verify that the unit is suitable for processing or takes the necessary steps to render it suitable. For example, as described above, the unit needs to be at the proper temperature (e.g. 19-25°C) . Also, original collection of the red cells in a suitable storage solution may be verified. If, for example, a two-part storage solution is used as described above, verification will be required that the red cells have been collected in the correct first part solution. The operator or attendant may also verify that the unit being treated (1) has the appropriate volume of red cells to qualify under such standards as may be applicable, (2) is leukocyte- reduced via collection or filtration, (3) has been appropriately concentrated, with the other components (plasma and platelets) removed and/or (4) is being processed timely (within five days after collection, for example) . [000190] Each qualification or verification is preferably recorded directly into an electronic data management system, via scanner, keyboard or other input device. That information may be stored in the data system memory in association with a unique number, code or other identifier that is associated with the particular quantity or unit of red cells being processed. For example, each unit or bag of red cells may have a unique bar code label, imbedded electronic memory or processor chip or other device that is scanned or interrogated by the operator when initiating the qualification or verification procedure. The verification or qualification data is stored for that particular quantity of red cells on suitable computer data storage media such as magnetic memory, hard disc drive, compact disc or floppy disc, so that the processing history of a particular quantity of red cells may be archived and/or retrieved as needed.

[000191] After the operator has verified or qualified a particular quantity of red cells for treatment, and the verification or qualification has been appropriately entered into the data management system, the operator then preferably selects the appropriate disposable fluid circuit assembly to be used with the particular red cell treatment desired. Upon selection of the appropriate disposable fluid circuit assembly, the operator may enter identifying data for such assembly into the data management system, which preferably has the ability to check the operator's selection and to alert the operator if the operator has chosen a fluid circuit assembly that is intended for a different red cell treatment procedure or is otherwise not appropriate for the desired treatment procedure or for the specific red cells to be treated. [000192] Upon selection of the appropriate disposable fluid circuit assembly, the operator attaches the particular quantity or unit of red cells. Preferably the connection is made using a standard sterile connection device, although any other sterile connection device/procedure approved by the blood processing center concerned may also be used. The operator then proceeds to the mixing station 34, and there installs the disposable fluid circuit assembly into a reusable controller or control module 40, which may be one of several such reusable controllers generally centrally located, for example on a carousel 42, or otherwise arrayed for efficient loading and unloading.

[000193] As discussed above and as will be described more fully below, the reusable controller 40 automatically directs fluid flow through the disposable fluid circuit assembly or module to mix a treating agent, such as a red cell pathogen inactivation agent, with the red cells (or other biological fluid being treated) in the desired ratio and, if desired, to process the combined treatment agent and red cells through a mixer, such as a static mixer, to assure complete mixing of the treating agent and the quantity of red cells or other biological fluid. This process may include, for example, automated reconstitution of a pathogen inactivation agent and/or an optional quenching agent, precision combination of the reconstituted agent (s) with the red cells, mixing of the combined red cells and treating agent (and optional quenching agent) , and directing the mixed red cells and pathogen inactivation agent to a storage or incubation container. Incubation and adsorption processes could also be accommodated at this station if desired. However, the extended time associated with incubation and adsorption lends itself to separate processing in order to free up the automated mixing device for treatment of additional units of red cells. [000194] When the mixing of the treating agent and the red cells is complete, the container of treated red cells (which may have a pre-attached adsorption container and storage container for later use) are sealed and severed from the rest of the automated fluid circuit assembly and taken to the incubation/adsorption station 36. At the incubation/adsorption station, the treated red cells are incubated for the desired period of time and at the desired temperature to assure that the treating agent has time to react with the red cells and effect the desired treatment. As noted above, the incubation time for the illustrated process will normally be about 12 hours and take place at a temperature of approximately 19-25°C, with gentle agitation keeping the red cells suspended and enhancing the treatment. The instrument or apparatus used for incubation or the operator should preferably record the time that incubation has started and the time of completion and record such information in the data management system.

[000195] After incubation, the treated red cells are then expressed, manually or automatically, through an adsorption device or into a container containing an adsorption device which adsorbs any unreacted treating agent, quenching agent or reaction by-products from the collected red cells. Other adsorption means may also be used.

[000196] When a container with an internal compound adsorption device is used, the treated red cells may remain in the container with the compound adsorption device for at least approximately 8 hours at 19-25°C, during which time the container will preferably be gently oscillated or agitated to assure contact with the adsorption device while avoiding undesirable hemolysis of the red cells. Again, the initiation and completion of the adsorption cycle are preferably recorded either manually or automatically in the data management system. After the adsorption is complete, the red cells may be transferred to a third container for storage or allowed to remain in the adsorption container, as desired.

[000197] From the incubation stage, the treated red cells are removed to the final processing/discharge station 38. There, an attendant or operator, after expressing residual air from the red cell storage container (for example, into an attached empty container) , seals and severs the treated red cell container from the rest of the disposable fluid assembly, if any, still attached. The operator or attendant may also prepare any samples of red cells for subsequent testing or storage as may be required by the practice or standards of the particular blood center or institution doing the processing. The operator also may review the process data for the treated unit or quantity of red cells to assure that the predetermined operation or protocol for treatment of the red cells has been carried out. Each of these checks or verifications also may be entered into the data management system so that each treated quantity of red blood cells has a complete treatment and verification history recorded in the data management system. The treated red cells may then be released for transfusion to the patients as required, with confidence that the particular procedure, such as pathogen inactivation, has been fully and completely carried out and that all necessary processing steps and ( conditions of the process or protocol have been met .

[000198] Figure 4 shows an alternative processing cell or production facility, having additional automated mixing devices or controllers 40 and greater capacity than the production cell shown in Figure 3. As with the production cell of Figure 3, the production facility or cell shown in Figure 4 employs a plurality of receiving/preparing stations 32, mixing stations 34 employing several carousels of reusable controllers 40, a larger or higher capacity incubation/absorption station 36 and a plurality of final processing/checking stations 38. Again, the stations are generally sequentially arranged and the controllers are centrally located for efficient loading and unloading.

DISPOSABLE FLUID CIRCUIT MODULE AND REUSABLE CONTROL MODULE [000199] Turning now to a description of the method and apparatus of the present invention employed in reconstituting and mixing the treating agent with the red blood cells, Figure 5 shows an operator loading or installing a disposable fluid circuit assembly or module 44 into the reusable controller or control module 40 of the present invention which controls flow through the fluid circuit assembly. Loading takes place, of course, after the quantity, e.g., container or bag, of red cells to be treated is attached to the disposable fluid circuit assembly. The disposable fluid circuit 44, which will be discussed in more detail later, preferably comprises a generally rigid tubing organizer or housing 46 with inflow tubing 48 connected to a container or bag of red cells 50 and inflow tubing 52 connected to a container of reconstitution liquid 54, such as the second part (e.g., dextrose) of a two- part solution, and an outlet tube 56 connected to an incubation/adsorption container or container set 58.

[000200] The reusable controller 40 may include a receiving station such as slot 60, into which the rigid tubing housing 46 is inserted. The controller 40 also preferably includes weigh stations 62, 64, and 66 for the red blood cell container 50, the reconstitution liquid container 54 and incubation container 58. Each weigh station includes a weigh scale on which the respective container rests and the weigh scales are used to control flow rates during processing, to detect leakage and to monitor the progress of the process.

[000201] Figure 6 diagrammatically shows the fluid flow path, valves, mixers, sensors, pumps and containers employed in the combination disposable fluid circuit module and control module as employed in carrying out pathogen inactivation in red cells, in accordance with one embodiment of the invention. The components shown in the dashed outline in Figure 6 generally comprise the portions of the fluid circuit assembly that are contained within the tubing organizer housing and those portions of the controller that interface with the fluid circuit to control flow therethrough.

[000202] As illustrated diagrammatically in Figure 6, the assembled system includes the red blood cell container 50, the reconstitution liquid container 54 and the incubation and/or compound adsorption container 58. The red cell container 50 communicates with the fluid circuit assembly via tubing 48 that defines the red cell inlet into the disposable fluid circuit assembly. Similarly, the container of reconstitution liquid 54 communicates via reconstitution fluid tubing 52 that defines an inlet for the reconstitution fluid into the fluid disposable circuit module. Incubation container or incubation/adsorption container or container set 58 communicates with the disposable fluid circuit assembly via tubing 56. Typically, the entire fluid circuit module shown in Figure 6, with the exception of the red blood cell container 50, is preassembled, and is provided to the user as a closed sterile assembly or kit. As indicated above, the red cell container is joined to the tubing 48 by a sterile connection device, whereby the entire fluid circuit module defines a closed sterile flow path for the processing of the red blood cells.

[000203] In the illustrated embodiment, the disposable fluid circuit module includes a sealed container having an interior chamber containing a treating agent, such as vial 68 containing a pathogen inactivation agent, and, optionally, a sealed container having an interior chamber containing a quenching agent, such as vial 70. The rigid housing 46 contains or mounts access members, not shown in detail in Figure 6, such as piercing members for piercing the septums of vials 68 and 70, and a fluid pump, which is preferably but not exclusively in the form of a syringe or piston-cylinder pump 72.

[000204] Tubing 52 defines a flow path between the reconstitution fluid container 54 and an inlet end of the syringe pump 72. Vial 68, and optionally vial 70, are connected to the reconstitution fluid flow path between the reconstitution fluid container 54 and the syringe pump 70. The reconstitution fluid flow path also may include a frangible member 74 (or other flow control member) immediately downstream of the reconstitution fluid container 54 to prevent flow of reconstitution fluid until the frangible member has been broken, thereby opening the flow path to fluid flow through the tubing 52. Pinch valves 76, 78, 80 and 82 control flow of reconstitution fluid, pathogen inactivation agent and optional quenching agent through their respective flow paths to the syringe pump 72.

[000205] Turning to the red cell flow path, the red blood cells communicate through the red blood cell tubing 40 and damping chamber 84 to the mixing junction or Y 86. A frangible flow control member 88 normally blocks flow until opened. The damping chamber accumulates a quantity of red cells to provide a constant flow of red cells to the mixing Y 86 and to smooth-out or dampen any pulsations or variations in the flow rate created by red blood cell pump 90 which, in a preferred embodiment, is a peristaltic pump located on the controller.

[000206] To deliver reconstituted pathogen inactivation agent to the mixing Y 86, the fluid circuit includes a tubing segment 92 extending between mixing Y 86 and junction 94 in the fluid flow tubing 52 that extends between the reconstitution liquid container 54 and the syringe pump 72. The tubing segment 92 may also include a damping chamber 96 to smooth out any flow variation in liquid flowing from the syringe pump to the mixing Y.

[000207] For conveying combined red cells and pathogen inactivation agent to the incubation container 58, the fluid circuit includes a flow path or tubing segment generally at 98. This flow path may include one or more static mixers or mixing sections generally at 100, for mixing the combined red blood cells and reconstituted pathogen inactivation agent (and any optional quenching agent) . As discussed in more detail later, this static mixing section may include one or more static mixers.

[000208] One form of static mixer that may be employed in the illustrated fluid circuit is the static mixer 102, which uses a series of alternating helical segments that repeatedly divide and rotate the fluid stream so as to provide a high degree of mixing of the combine fluid stream. Because this type of static mixer tends to mix the fluids streams in a generally radial direction and may, in highly unusual circumstances, allow some portion of the fluid stream to remain insufficiently mixed, axial mixer 104 (which, in general, is also a static mixer) may be provided between the other static mixers 102 to mix the combined fluid stream in a non-radial direction so as to better assure a total and complete mixing of the combined red cell and pathogen inactivation agent fluid streams. Pinch valve 106 may be provided to control flow between the mixing Y 86 and the incubation container 58, i.e., through tubing segment or flow path 98. [000209] In general, fluid flow through the fluid circuit assembly or module 44 is controlled by the controller or control module 40, operating through pinch valve 76-82, 108, 110 and 106, red blood cell pump 90, syringe pump 72 and a frangible breaking mechanism (not shown in Figure 6) for opening the frangible members 74 and 88 to allow fluid flow through the reconstitution fluid and red blood cell flow lines. Additionally, the controller may have sensors 112, 114 and 116 located, respectively, in the fluid flow path entering the syringe pump, in the red blood cell flow path between the damping chamber 84 and mixing Y 86 and in the combined fluid flow path downstream of the mixing Y 86. These sensors allow the controller to sense fluid flow at those locations for controlling the sequence of operation and sensing potential departures from the desired process or protocol. Additionally, although not shown in Figure 6, the controller may include a load cell on the syringe pump actuator to monitor the force exerted by the syringe pump and/or syringe pump position. By monitoring the force exerted on the syringe, the controller is able to identify when the syringe plunger bottoms out within the barrel or cylinder of the syringe for future reference. In addition, the load cell may provide a force indication or signature during the disposable fault check, indicate a force profile or signature during the reconstitution cycle and provide for potential fault detection during other processing.

DISPOSABLE/FLUID CIRCUIT ASSEMBLY AND FLOW ORGANIZER HOUSING [000210] One embodiment of the disposable fluid circuit assembly 44 is shown in Figures 7-13. It includes the rigid tubing organizer housing 46, which houses many of the components of the fluid circuit assembly, red cell inflow tubing 48, reconstitution inflow tubing 52, outlet tubing 56, incubation container 58, adsorption container 118 and displacement air container 120. As best seen in Figures 8-12 (See Fig. 12 in particular) , the portion 122 of the fluid circuit that is contained in or by the housing 46 is arrayed on a fluid circuit organizer 124. As seen in Figures 10 and 12, the disposable fluid tubing circuit portion 122 includes the red cell inflow tubing 48 and reconstitution fluid inlet tubing 52, with respective frangible flow control members 88 and 74. The reconstitution fluid tubing extends past the frangible flow control member 74, to the inlet end of the syringe pump 72. Vials 68 and 70 are connected via tubing segments 68t and 70t to the reconstitution flow tubing at connectors 68c and 70c.

[000211] Red cells enter the disposable tubing assembly through the red cell tubing 48. The red cell flow path continues, past the frangible flow control member 88, to the damping chamber 84 and from there to the mixing Y or junction 86. It is at the mixing Y where the red cells are combined with reconstituted treating agent and optional quenching agent from the syringe pump. Tubing segment 126 connects the syringe pump, via damping chamber 96, with the mixing Y 86. [000212] From the mixing Y, the combined red cell and treating agent fluid stream flows through tubing to a first static mixer 102, axial mixer 104 (also a type of static mixer) and a second static mixer 102. From there, the mixed stream flows through exit tubing 56, which leads to the incubation container 58 (not shown in Figures 9 and 10) . [000213] The disposable fluid circuit assembly 122 is arrayed on the fluid circuit organizer 124 as best seen in Figures 10 and 12. The fluid circuit organizer 124 (see Figures 12 and 13) is a generally rigid plastic plate with pre-formed slots and recesses defining specific locations into which the various parts of the disposable tubing assembly 122 are placed. For example, the organizer has a slot 128 for receiving red cell inflow tubing 48, slot 130 for receiving reconstitution fluid flow tubing 52, slot 132 for receiving outlet tubing 56, and slots 134 and 136 for receiving static mixers 102. The organizer also includes recess 138 for receiving the damping chamber 84, recess 140 for receiving the mixing Y 86, and recess 142 for receiving axial mixer 104, as well as other slots and recesses for other tubing segments and components of that portion of the disposable fluid circuit 122 that mounts on the organizer.

[000214] The organizer 124 also may have such number and location of pinch valve sites as needed to control flow through the fluid circuit. , As shown, the organizer includes pinch valve site 76 on the reconstitution liquid inflow line, and pinch valve sites 80 on the treating agent flow line 70t and 82 on the optional quenching agent tubing segment 68t. Pinch valve site 78 is located on the reconstitution fluid inflow line, between the connectors 68c and 70c (where the optional quenching agent and treating agent enter the reconstitution liquid flow path) . Pinch valve site 108 is located along the reconstituted treating agent tubing segment 92, and pinch valve site 110 is located in the red cell inlet flow path, between the damping chamber 84 and mixing Y 86. [000215] At each pinch valve site of the organizer 124 the tubing of the fluid circuit extends across a diagonal pinch valve surface 144 which cooperates with pinch valve plungers located in the reusable controller. The respective tubing extends generally transversely across each pinch valve surface, as seen for example in Figure 10. Control of fluid flow through the respective tubing segment is controlled by actuating the plunger of the controller to pinch the respective tubing against the pinch valve surface 144 to close or block flow through the tubing or to retract the pinch valve plunger to release the tubing and allow the flow therethrough (as seen in Figures 14 and 15) .

[000216] After the tubing assembly 122 is assembled onto the organizer 124, a plastic overwrap or pouch 146 is attached, which fully encloses the tubing organizer and associated fluid circuit assembly to provide additional containment of the fluid circuit in the unlikely event of any leakage from any of the components mounted on the organizer. The overpouch or overwrap generally comprises a pair of facing plastic sheets of any suitable material that is preferably flexible and heat sealable or weldable. The sheets are sealed together along seal in 148 around the perimeter of the tubing organizer. For complete containment, the overpouch is sealed to the tubing 126 extending from the syringe pump, as well as to the reconstitution liquid, red cell and exit tubings 52, 48 and 56, and the tubing or connectors extending from the vials 68 and 70.

[000217] After the tubing organizer 124 and tubing assembly 122 are sealed within the overpouch 146, they are mounted between the rigid shells of the organizer housing 46. The organizer 46 may have registration openings 150 that mate with corresponding registration projections 152 (see Figure 9) in the rigid housing to assure proper positioning of the tubing organizer and associated parts within the housing. [000218] As may be seen in Figures 8 and 9 the rigid tubing organizer housing 46 includes a syringe pump cavity 154 which captures the barrel of the syringe pump between flanges 156 located on the syringe barrel 158. This allows the syringe plunger 160 to extend outside of the housing 46 for actuation by the controller 40, as will be discussed later. [000219] The housing 46 also includes regions 162 and 164 for receiving the vial containment vessels 166 and 168 that contain the treating agent vial 68 and, optionally, the quenching agent vial 70. Regions 162 and 164 are open at the upper end to receive vial access actuators (not shown) associated with the controller for accessing the contents of the vial. As will be described in more detail later, the vial actuators comprise plungers which depress the vial containment vessels and force a piercing member or spike through the vial septum so that the contents or inner chamber of each vial communicates with each respective fluid flow path 68t and 70t. [000220] As best seen in Figures 7 and 8, the frangible members 74 and 88 are preferably located in a window 170 defined in the housing 46. The frangible members are in a spaced apart, parallel relationship within the window. The controller 40 includes a frangible breaking apparatus which, upon appropriate command from the controller, breaks the frangible connectors preferably, but not necessarily, one at a time, to open fluid flow through the respective tubing. The details of the frangible connector and breaking apparatus are discussed hereinafter.

[000221] For ease of handling, the rigid tubing organizer housing includes a large opening 172 defining a handle 174 that may be grasped for inserting the tubing organizer housing into a receiving station on the controller.

SYRINGE PUMP [000222] Turning now to a more detailed description of various components of the fluid circuit assembly, attention is first directed to the syringe pump 72, which is shown in more detail and Figures 16-19. Figure 16 is a side view of the syringe pump 72, showing the outer barrel or cylinder 158, plunger 160 and a containment sleeve 176. The barrel is generally cylindrical, and is open at one end for receiving the plunger 160 and has an inlet/outlet port 178 at the other end. The plunger includes a pair of spaced apart radial flanges 156 that cooperate with the rigid tubing organizer housing to hold the syringe pump in place. Flange 156a is located at the plunger-receiving end of the barrel and flange 156b is located between the ends of the barrel. [000223] As best seen in Figure 17, Plunger 160 mounts a resilient cap or piston 180 at one end, which is slidably received within the cylindrical chamber of the barrel 158. The other end of the plunger is attached, such as by threads, to a plunger cap 182 which is engageable by a cooperating mechanism on the controller 40 for reciprocating the plunger within the barrel. The containment sleeve 176 surrounds the portion of the plunger that extends from the barrel to provide secondary sealed containment of the syringe pump in the event any liquid leaks past the plunger piston 180. The containment sleeve has radial end flanges 184 for sealed attachment to the upper flange 156a of the barrel 158 and to the plunger cap 182, so as to enclose the plunger portion that extends from the barrel .

[000224] The containment sleeve 176 is axially extendable to accommodate movement of the plunger into and out of the syringe barrel. Different configurations may be employed to accommodate axial extension and contraction of the containment sleeve. In the illustrated embodiment, the wall of the containment sleeve is cross sectionally undulating or corrugated so as to extend or contract in a bellows or accordion-like fashion, as shown for example in Figures 18 and 19. The sleeve may also be of resilient material that stretched to allow plunger extension. Figure 18 shows the pump 72 with the plunger 160 fully depressed into the barrel or cylinder 158, and Figure 19 shows the pump with the plunger fully retracted, illustrating the containment sleeve fully surrounding the portion of the plunger extending from the barrel in both positions.

[000225] For accuracy, it should be noted that in the embodiment of the syringe pump illustrated in Figures 18 to 19, specifically, the containment sleeve 176 does not terminate in radial flanges 184, but terminates in axially- extending ends that are sealingly attached to the plunger cap 182 at one end and to a barrel cap 182 at the other end. The barrel cap 186 is attached to the open end of the syringe barrel and may include an extension 188 for receiving and guiding the plunger 160. Raised ribs or grooves 190, 192 defining detents in the extension and/or the plunger may be provided for indicating or limiting the stroke of the plunger.

VIAL ASSEMBLY [000226] Turning to the vial assembly, Figures 20-23 show a vial assembly that may be employed in this illustrated embodiment of present invention. The vial assemblies for the red cell treating agent (e.g. pathogen inactivation agent) and optional quenching agent are the same, and only red cell treating agent vial assembly will be described in detail. [000227] The vial assembly, as shown generally Figure 20, includes a vial containment vessel 166 for double containment of the vial and an access assembly. The features of the vial assembly are more easily understood with reference to Figures 21-23. Figure 21 is an exploded perspective view and Figures 22 and 23 are cross-sectional views, showing the vial assembly before the contents of vial are accessed (Figure 22) and after the contents are accessed (Figure 23) .

[000228] As shown in Figures 20-23, the vial 68 is contained within a vial housing 166. The vial 68 is a standard vial with glass or plastic body 194 open at one end, which opening is closed by a pierceable septum 196 of latex or other suitable material. The vial is received in the vial containment vessel 166 with the septum facing the access assembly 198. The open end of the containment vessel 166 is generally covered by a pierceable diaphragm 200 that is peripherally captured and sealed between the end of the containment vessel and the access assembly 198.

[000229] The access assembly 198 comprises a spike or piercing member body 202 and a cap structure 204 that is attached to the containment vessel 166 at one end and slidably received within the spike body 202 at the other end. As may be seen in Figure 22, a center spike or piercing member 206 is located within the spike body 202. The spike body includes an outer cylindrical wall 208 spaced from and surrounding at least a portion of the spike. The space between the center spike 206 and peripheral wall 208 defines a receiving slot for slidably receiving a portion of the cap structure 204 (See Figure 23) .

[000230] Still referring to Figures 22 and 23, the cap structure 204 has a base portion 210 attached to the vial containment vessel 166, and a projecting cylindrical wall structure having a generally cylindrical outer wall 212 and generally cylindrical inner wall 214. The inner wall forms a spike receiving passageway 216. The inner wall terminates in proximity to the pierceable diaphragm 200, and is closed by a pierceable cover 218.

[000231] To access the contents of the vial, the vial 68 and access assembly 198 are pushed together, forcing the piercing member or spike 206 through the cover 218, through the pierceable diaphragm 200 and through the vial septum 196 as shown in Figure 23. The spike body 202 has a plurality of axially extending slots 220 that allow the outer wall 208 to flex slightly outwardly as the spike is inserted into the vial . The upper end of the outer wall terminates in an inwardly extending hook 222 that resiliently snaps into annular slot 224 in the outer wall 212 of cap structure 204 and prevents inadvertent separation of the vial and spike when the spike is fully inserted into the vial . As seen in Figure 22, the hook 222 is received in a lower slot 226 in the outer wall 212 when in the non-vial accessing position. The tapered upper surface of the hook releases the hook from slot 226 when the spike body is pushed toward the vial. 0-ring 228 provides a seal between the spike 206 and the inner wall 214 of the cap 204. As noted later, the flow path leading to the hollow spike preferably is pressurized when the hollow spike is inserted into the vial to force or blow any treating agent away from the orifice and prevent drawing any unreconstituted agent unto the spike.

STATIC MIXER [000232] As described above, the disposable fluid circuit assembly or module includes at least one static mixing section to achieve thorough mixing of the pathogen inactivation fluid (or other treating agent) with the concentrated red blood cells (or other biological fluid) . In the illustrated embodiment, (see Figures 6 and 9) the static mixing region includes a first mixer 102, a second mixer 104 and a third mixer 102, identical to the first. While this may be the preferred number and order of the mixing sections, in keeping with the present invention the first mixing section may be used alone, if it is so desired, and still provide a very high degree of homogeneity in the resultant mixture. It is also in keeping with the present invention to employ the first mixing section in combination with a second or additional mixing sections to provide a greater degree of confidence that the sufficient mixing has occurred.

[000233] The first static mixer 102 is best seen in Figure 24. As shown in that figure, which illustrates an exemplary portion of the first static mixer 102, the static mixer comprises a tubing or flow path segment that contains at least one and preferably a plurality of mixers or mixing elements 230 serially arranged within the flow path to mix the fluid stream as it flows along the combined fluid flow path, without the need for any moving parts or external forces, other than the force of the moving stream itself. Each mixing element 230 preferably comprises a generally planar surface that is shaped to divide the fluid stream and to rotate the fluid stream as it passes by the mixing element. In this regard, the mixing element has a substantially helical or auger shape. The shape of the element may also be analogized to a ribbon in which one end is rotated 180° relative to the other end.

[000234] The leading edge 232 of the mixing element extends diametrically across the fluid stream so as to divide or bisect the fluid stream into two sub-streams. As a result of the generally helical or auger shape of the mixing element, each sub-stream is twisted or rotated through 180° as it moves past the mixing element. The side edges of the mixing element are closely positioned against the inside wall of the flow path (e.g. flow tubing), so that essentially all of the fluid is constrained to move in the manner directed by the mixing element. The mixing elements do not intentionally rotate, but are preferably stationary to force the fluid to move around them.

[000235] As illustrated, there is a series of mixing elements 230 disposed one behind the other in the fluid stream. The leading edge 232 of each succeeding or downstream mixing element is preferably located at 90° relative to the leading edge (and following edge) of the next most preceding or upstream mixing element so as to divide the stream in a different direction than originally divided. In addition, as illustrated in Figure 24, each succeeding mixing element is preferably shaped to rotate or twist the fluid stream in the opposite direction as compared to the preceding (upstream) or the following (downstream) mixing element. So, as illustrated in Figure 24, the first mixing element bisects the fluid stream vertically and rotates or twists each sub-stream in a clockwise direction. This may also be referred to as a right- hand twist. The leading edge of the next mixing element extends in a generally horizontal direction to subdivide the fluid stream in a direction different from that by the first mixing element and, additionally, has a left-hand or counterclockwise twist so as to rotate or twist the mixture sub-streams in a counterclockwise direction. After passing the second mixing element, the fluid stream then engages a mixing element identical to the first mixing element with a vertical leading edge and a clockwise or right-hand twist. Any number of mixing elements may be employed to obtain the desired degree of mixing. For achieving a high degree of homogeneity it may be preferred to have ten or more and preferably twelve to thirteen or more mixing elements employed in the first static mixing section. If only a single such static mixer 102 is employed, it may be desired to have as many as 24 and possibly more such mixing elements. [000236] The alternating clockwise-counterclockwise arrangement of mixing elements 230 described above in the first mixer 102 of Figure 24 produces rapid mixing and creates a level of turbulence in the fluid stream which further encourages mixing as the fluid streams pass through the first mixing section. Alternatively, the mixing elements may be arranged to turn or twist the fluid in the same direction to provide a more laminar mixing flow with less pressure drop through the mixing section. Such an arrangement is illustrated in Figure 25, which shows a portion of a mixing section having three mixing elements 230, each mixing element having the same shape (right-hand twist) so as to rotate or twist the fluid stream in the same direction. As seen in Figure 25, the leading edge 232 of the first element is vertical, bisecting the fluid stream and twisting or rotating the fluid stream in a generally clockwise manner. The next mixing element is positioned so that its leading edge is horizontal, to further sub-divide the fluid stream, but also to rotate or twist the fluid stream in a clockwise direction, leading the fluid stream to the next or third mixing element which has vertical leading edge and is similarly positioned or shaped to rotate or twist the fluid stream in a clockwise direction. A similar number of mixing elements may be provided as in the mixer fashioned in accordance with Figure 24. However, additional mixing elements may be desirable in the Figure 25 embodiment, as compared to the Figure 24 version, since the Figure 25 version employs a generally laminar flow through the mixing section and has less turbulence than in the Figure 24 static mixer.

[000237] The static mixer described above, in effect subdivides each unit of the fluid stream in what may be described as a stretching and folding action, and the mixing may be defined by the number "2ⁿ, " where "n" represents the number of mixing elements. For example, with 13 such mixing elements and fluid stream transformations, 2ⁿ = 1.6 x 10⁴, and a hypothetical 1 cm fluid stream segment is repeatedly divided and subdivided so that it becomes less than 1 μm. If a single static mixing section as described above is employed, it may include approximately 24 mixing elements for enhanced mixing. It is further understood that mixing may be optimized by flowing the fluid stream in a laminar flow region or condition with a Reynolds number less than 100. When mixing red blood cell concentrate with pathogen inactivation agent such as described above it is desirable to obtain thorough mixing of the agent to a concentration from about 0.150 to 0.300 mM in the final mixed fluid stream product.

[000238] One particularly suitable mixing element 230 that may be employed in the static mixers as shown in Figures 24 and 25 is available as the Kenics KM static mixer from Chemineer, Inc. of Dayton, Ohio. These static mixing elements are modular, allowing any number desired to be placed in the fluid stream to achieve the desired mixing. In addition, these mixing elements are available in a wide variety of materials such as polyvinyl chloride or other plastics that may be suitable for use in a disposable medical fluid circuit, as described herein.

[000239] The arrangement and shape of the mixing elements 230 in the first static mixer 102 results in a mixing of the fluid streams mainly in a radial direction. It may be possible in certain unpredictable circumstances for minute fluid streams to pass through the first mixing section without being mixed sufficiently to provide the highest levels of homogeneity that may be desired, for example, when mixing a pathogen inactivation agent with biological solutions such as blood or blood components . Accordingly, the combined fluid flows streams may include the axial static mixer 104, which is adapted to reshuffle the fluids and to mix the fluid in a direction other than or in addition to radial, such as mixing axially relative to the direction of flow. Such a mixer may be of a variety of shapes or configurations. As shown in Figures 6-12, for example, the axial static mixer 104 is a simple enlarged accumulation chamber or reservoir into which the combined fluids flow into an upper opening in the chamber. When the fluid stream enters the chamber, it disperses in a variety of directions other than radial (not excluding some radial dispersion as well) . Any unmixed fluid streams from the first mixer are thus broken up and reshuffled or redistributed within the axial mixer.

[000240] Although the mixing section could terminate at the end of either the first mixer 102 or the second mixer 104, to provide an even greater degree of confidence with respect to the homogeneity of the resulting mixture, and to further process any unmixed streams that are broken up in the axial mixer 104, a third mixer 102 may be provided. In the illustrated embodiment, the third mixer 102 is essentially identical to the first mixer 102. It comprises at least one and preferably a plurality of mixing elements 230 which are serially located along the fluid flow path and shaped to repeatedly divide and twist or rotate the fluid flow path in the same or opposite directions as the fluid passes through the third mixing section. The number of mixing elements may be varied, but ten or more, and twelve to thirteen mixing elements are preferred in each of two separate mixing sections 102.

[000241] From the third mixer 102, the mixture flows preferably into the incubation/adsorption container or container set, where it may be further processed. It should be appreciated that one benefit of the system illustrated above, is that a closed medical fluid circuit assembly is provided that results in precise addition of a pathogen inactivation agent, or other fluid, with a blood component, or other medical fluid, and thorough mixing to provide an extremely high level of reliability that the resulting mixture is sufficiently homogeneous and that the pathogen inactivation agent or other treating agent is sufficiently mixed with the red blood cells (or other biological fluid) to carry out the pathogen inactivation (or other treatment) process. [000242] Additional discussion of the static mixers described above and alternative static mixing arrangements that may be employed is found in U.S. Patent Application Serial No. 60/435,146 filed December 20, 2002, entitled, "Static Mixing Apparatus and Method" , which is incorporated by reference in this description.

REUSABLE CONTROLLER [000243] Figure 5 illustrates the installation of the disposable fluid circuit module 44 into a reusable controller 40. The reusable controller as shown in Figure 5 includes the receiving station or slot 60 into which the rigid tubing organizer housing 46 is inserted. As will be discussed in more detail, the controller includes apparatus associated with the receiving slot which act upon the various valves, pump and vials and frangible members mounted on or in the organizer housing 46 to control processing of fluids through the disposable fluid circuit module. In addition, as can be seen from the face of the controller, shown in Figure 26, the controller includes weigh station 62 for receiving the container of concentrated red cells 28; weigh sta'tion 64 for receiving the reconstitution liquid container 54; and weigh station 66 for receiving the incubation container 58. [000244] Each station in the illustrated embodiment is formed as a recess or pocket in the face of controller 40, allowing the respective container or bag, which is typically of flexible plastic material, to be placed in the recess in a lay-flat position. As best seen in Figure 38, the bottom surface or floor of each station may comprise a weight scale 233 of suitable conventional or other design which is connected to the controller processing system, so that the weight of the containers (and changes in the weight) can be detected and used to calculate flow rates, to monitor for compliance with the processing protocol, to detect potential leakage, and to track other processing parameters and progress .

[000245] A peristaltic pump 234 for controlling the flow of concentrated red cells into the disposable fluid circuit processing system is also located on the face of the controller. As best seen in Figure 27, the pump comprises a rotor 236 with a plurality of rollers 238 that compress the tubing. The pump has a hinged curved door 240 that encloses the rotor and acts as a platen, against which the red cell inlet tubing 48 is compressed by the rollers of the pump. If desired, a special pump tubing segment 242 made of silicone or other material may be used in the inlet tubing for contact with the pump rotors .

[000246] Figure 26 is a frontal view of the illustrated reusable controller, with the disposable fluid circuit assembly installed therein. It should be noted that the illustrated controller is merely one configuration that may be used in accordance with the present invention. As seen in Figure 26, the controller operates with one fluid circuit assembly at a time. As will be discussed later, the controller may include a pair of adjoining slots or additional disposable receiving slots for receiving two or more tubing organizer housings and associated containers so that two or more units of red cells may be processed simultaneously through separate disposable fluid circuit assemblies. Also, the controller shown in Figure 26 employs a slot-type receiving station for the tubing organizer housing. Alternatively, as will also be discussed later, the tubing organizer housing could be mounted flat on the face of the controller or mounted behind a door located on the face of the controller, or other suitable mounting arrangements may be used.

CONTROLLER ACTUATOR ASSEMBLY [000247] Figures 28 and 29 are perspective views of actuator assemblies in the controller 40 that operate to control fluid flow through the disposable fluid circuit. In the illustrated embodiment, the controller includes a valve actuator assembly 244, a frangible element breaker assembly 246, a pump actuator assembly 248 and a vial actuator assembly generally at 250. The actuator assemblies are mounted to a slot housing 252 that forms the receiving slot or station 60 for the rigid organizer 46.

VALVE ACTUATORS [000248] The valve actuator assembly 244 is best viewed in Figures 34-36. The assembly comprises a base plate 254 and a number of valve actuators 256 corresponding to the number of pinch valve positions on the tubing organizer housing 46. As presently contemplated, there will be a plurality, up to about seven such valve actuators, with the illustrated embodiment using five actuators. The valve actuators are assembled to the mounting plate and the mounting plate is attached at the desired location to the slot housing 252. The mounting plate locates the valve actuators in positions aligned with the pinch valve sites in the disposable tubing organizer housing 46, which are accessible through aligned openings in the slot housing.

[000249] The valve actuators are preferably electromechanical linear actuators that pinch the respective tubing closed by extending an actuator rod 258 toward the disposable fluid circuit housing. The end of the rod pushes against the tubing and compresses it against a pinch valve surface 144, as described earlier, on the tubing organizer 124. Each valve actuator is preferably an electro-mechanical, direct-driven device, not requiring linkages or motion-transferring components. The position, speed, acceleration and position of the actuator are preferably software controllable for monitoring and control, if desired, by the controller operating system. Such actuators are commercially available from Alpha Gear Drives Corp., of Elk Grove Village, Illinois. [000250] In the illustrated controller, it is contemplated that the normal or home position for each valve actuator is fully retracted, with the respective tubing being in open flow condition. When the actuator receives a signal from the controller, it extends the actuator rod until it encounters a sufficient resistance to further advancement, for example, 10 pounds resistance force, which indicates complete pinching. At this point, the controller signals the actuator to stop advancing the actuator rod and to hold it in (the pinching or closed) position until it receives a signal to retract to the home (or open) position.

FRANGIBLE BREAKER [000251] The frangible element breaker assembly 246 preferably comprises a dual motion actuator 260 mounted on a base 262 for mounting on the slot housing 252. When mounted on the slot housing 252, the actuator is aligned with the window 170 formed in the fluid organizer housing 46, in which the frangible elements 74 and 88 are located. As with the valve actuators, the actuator 260 is preferably electromechanical and, as illustrated in Figures 30 and 31, preferably has two directions of motion - axial and rotary - for an actuator rod 264. The actuator rod 264 carries, at its distal end, a frangible breaker head 266. The breaker head 266 has a generally flat configuration with a frangible- receiving slot or groove 268 or alternatively a pair of opposed slots or grooves.

[000252] A typical frangible flow control member is illustrated in Figures 83 and 84. The frangible member 270 is sealed within flow tubing 272 to block flow through the tubing until opened. The frangible member is opened by bending it laterally, causing it to break out a line of weakness (see Figure 84) , opening a passage through the tubing. [000253] When actuated, the breaker head 266 is parallel to the frangible connectors and in a retracted position. It is extended by the actuator rod 264 to a position between the frangible members . The actuator then rotates the breaker head 90 degrees, to the position shown in Figure 30 in which a selected frangible member is located within the slot 268 in the breaker head. When signaled by the controller, the actuator retracts the actuator rod, pulling the breaker head 266 and exerting lateral force on the frangible member. The ends of the frangible member are held against lateral movement by the rigid housing 46, and the force is exerted on the intermediate weakened area, breaking the frangible member and opening its respective flow path to fluid flow. The actuator rod is then rotated 90 degrees to release the broken frangible from the breaker head, and further rotated 90° in the opposite direction to carry out the same sequence on the other frangible member, or retracted to the home or start position. As noted above, the breaker head 266 may have a pair of opposed slots 268 to break both frangibles simultaneously, as shown in Figures 80 and 81.

[000254] Figure 82 illustrates an alternative frangible breaker that also employs an electromechanical actuator that rotates a breaker head 274 to break two frangible members simultaneously by bending the frangible members at the weakened area of the frangible to cause it to fracture, as seen in Figure 84.

PUMP ACTUATOR [000255] Turning now to the pump actuator assembly 248 shown on Figure 33, the pump assembly 248 comprises a linear electromechanical actuator 276 with a linear actuator rod coupled at the distal end to a pump connector 278. The distal end of the pump connector 278 includes a horizontal and vertical slots for receiving the radial flange 280 (located on the cap of the pump plunger) when the disposable tubing organizer housing 46 is inserted into the receiving slot 60 of the controller.

[000256] The pump actuator, in response to signals from the controller, reciprocates the plunger as needed to reconstitute the pathogen inactivation and quenching agents and to pump the reconstituted agents to the mixing Y 86 for mixing with the red cells to be treated. As will be discussed in more detail later, the pump actuator retracts the plunger to draw in the desired quantity of reconstitution agent, it then reciprocates the plunger to repeatedly inject reconstitution liquid into one of the vials 68 and 70 and withdraw the liquid to reconstitute the agent. Then the same process is carried out for the other vial. Following reconstitution, the valve actuator carefully meters the reconstituted agents to the mixing Y for mixing with the red cells. The pump actuator may also include a pump force and /or position sensor 282 to monitor the force exerted by the actuator and/or the positions of the actuator.

VIAL ACTUATOR

[000257] The vial actuator assembly 250 also comprises at least one and possibly two electromechanical actuators 284 mounted on a base structure 286 supported on the slot housing 252. The vial actuator includes a linear actuation rod which mounts plungers or pushers 288. The valve actuators 284 are mounted such that the plungers or pushers are aligned with the vial containment vessels 166, 168 when the disposable tubing organizer 46 is inserted into the slot. When actuated, the valve actuator pushes the plungers downwardly against the vial containment vessel, depressing the vials contained therein forcing the hollow spike 206 of each access assembly through the pierceable diaphragm 200 and vial septum 196 to access the contents of the vial. As previously discussed, the spike may be inserted so that the fluid openings in the distal of the spike are just inside the inner surface of the septum so as to enhance mixing of the reconstitution fluid with the pathogen inactivation or optional quenching agent.

[000258] A single actuator may be used to depress both vial pushers or alternatively the vial actuator assembly may include two actuators for independent actuation as illustrated, as desired.

A PROCESSING PROCEDURE [000259] When it is desired to carry out red cell treatment process as described herein and, in particular, when employed [000260] For red cell pathogen inactivation treatment employing a pathogen inactivation agent, referred to as S303, described above, and a quenching agent gluthione, referred to as GSH, the controller and the control system go through a series of steps. For purposes of this description, these steps are broken down into a start-up procedure, a preparation procedure, a reconstitution procedure, a dosing procedure and a completion procedure.

[000261] Briefly, the start up procedure comprises powering up the controller, positioning the actuators at the desired loading location and carrying out certain basic calibration checks for the weigh stations. In the preparation procedure, the controller prompts the user to load the disposable fluid circuit assembly into the controller and then carries out a series of tests and checks for leakage and the like before beginning the procedure. During the reconstitution procedure, the pathogen inactivation and quenching agents are reconstituted and the reconstituted pathogen inactivation agent and quenching agent are drawn into the syringe pump for subsequent injection into a stream of red blood cells drawn from a collection container. The dosing procedure includes mixing of the pathogen inactivation agent, quenching agent and red blood cells, passage of the combined fluid stream through the mixers and into a container (such as an incubation container) . The controller and system then go through a completion procedure culminating with sealing and severing of the incubation container, which contains the treated red cells, and instructing the user on removal of the disposable fluid circuit assembly from the controller, and preparing for the next treatment cycle. The specific steps of each procedure are set forth in the following table 1.

TABLE 1 START UP

PREPARATION

RECONSTITUTION

Pull air into syringe Fill the syringe with half of the volume of air required to purge dextrose out of the recon line from the incubation bag.

Close dose and mixer valves 174, 176

Open dextrose valve 146

Purge air from syringe The remaining air is purged from the syringe using the recon sensor to detect the fluid level in the syringe.

Close dextrose valve 146

Open dose and mixer valves The reconstitution circuit is vented to ambient (into the Incubation container) prior to reconstitution of the powders to prevent pressure buildup and to allow better force sensor performance .

Close dose and mixer valves 174, 176

Check dextrose volume used The dextrose bag volume is compared to the volume at the start of dextrose prime to determine how much is actually in the syringe. An error will occur if the correct amount is not available. The final syringe position is also checked.

Pressurize the recon circuit This ensures any movement through GSH valve, when it is opened, is toward the vial . For safety reasons, the system should keep the powder from leaving the vial when possible. This also allows a GSH valve occlusion check just prior to compression.

Open GSH valve 152 When the GSH valve is opened, the pressure in the recon circuit should force air into the vial, preventing any powder from leaving the vial. This also allows a GSH valve open check just prior to compression.

GSH recon compression stroke This step forces the cycle volume and the residual volume of dextrose into the GSH vial to

DOSING

CO PLETION

Ste Description

ALTERNATIVE EMBODIMENTS [000262] The present invention has been described in terms of one or more specific embodiments. .However, there are alternative that may also be employed without departing from the present inventions.

ALTERNATIVE CONTROLLER AND FLUID CIRCUIT ASSEMBLY [000263] Figures 39-60 show an alternative reusable mixing controller or control module 290 for carrying out, in its preferred form, pathogen inactivation in red blood cells, and an associated disposable fluid circuit assembly 292 that is adapted to be mounted on the reusable module for automated reconstitution and mixing of the pathogen inactivation or other treating agent with the red cells (or other biological fluid) to be treated. As may be seen in Figures 39-42, the mixing controller or control module 290 comprises a loading station 294, into which the disposable fluid assembly is mounted, a weigh station 296 for a red blood cell container, a weigh station 298 for a reconstitution fluid container, a weight station 300 for a treated red cell container, a peristaltic pump 302 for controlling flow of red blood cells into the disposable fluid circuit assembly, a tubing sealer 304 for sealing and severing plastic tubing and a display 306, such as liquid crystal display, touch screen or other output/input device, for user information and input. [000264] The weigh stations in this embodiment are accessible laterally for ease of loading. Specifically, as readily seen in Figure 39, the weigh station 300 for the treated red cells is open at the front and the right side, making manual loading of the container onto the weigh scale much easier than the front-load-only arrangement seen in controller 40 of Figure 5. [000265] Figure 41A shows the loading station of the controller with the door 310 in the open position. As may be seen there, the loading station comprises a recessed housing 900, sized to receive the fluid circuit assembly 292 in an upright face-on position (as distinguished from the edge-on receiving slot configuration of the earlier embodiment) . [000266] Viewing the inside of the housing as seen in Figure 41A, portions of the working ends of the actuator assemblies themselves may be seen, which engage the disposable fluid circuit assembly. The actuator assemblies themselves are substantially similar to those describe above, subject to reconfiguring or repositioning to accommodate the different position of the disposable in the controller 290 of Figure 41. More specifically, viewable in Figure 41A are at least pinch valve plungers 902, pump plunger engagement member 904 and frangible breaker 906. As can also be seen in Figure 41A, the pump plunger engagement member 904 is slotted at 908 to slidably receive an end flange of syringe pump plunger when the fluid circuit assembly is inserted. Also, slotted receiver 910 is located on the housing to slidably receive a radial flange located on a syringe pump barrel, similar to the syringe barrel flange illustrated in prior embodiments.

[000267] Figure 41B is a rear perspective view of the housing 900 with associated actuator assemblies mounted thereon. As with prior versions of the controller, this controller employs a vial actuator assembly or subassembly 912, valve actuator assembly or subassembly 914, frangible breaker assembly or subassembly 916 and pump actuator assembly or subsassembly 918. As noted above, these subassemblies employ electromechanical actuators as earlier described and differ from the earlier-described embodiment principally in reconfiguration or repositioning to accommodate the different position of the disposable fluid circuit.

[000268] Reconfiguration or repositioning is most evident in the valve actuator and pump actuator subassemblies. For example in the controller 290 (and associated disposable) the pinch valve sites are aligned in a row and thus, as shown in Figure 41C, the valve actuators 920 are mounted in corresponding alignment on a pre-drilled mounting block 922 that positions the actuator plungers to correspond to the location of the tubing pinch sites in the corresponding disposable, which will be described in more detail hereinafter.

[000269] The syringe pump actuator subassembly 918 is best seen in Figure 41D. This subassembly differs from that described earlier in connection with controller 40, in that the actuator 924 is indirectly connected to the plump plunger engagement member 904. As shown in Figure 41D, a horizontal connecting member 926 extends between and connects the actuator 924 to the plunger engagement member 904. The connecting member is slidably mounted to rails 927 on a base plate 928, which also mounts the actuator 924.

[000270] With the above arrangement, axial movement of the actuator shaft causes connecting member 926 to slide up or down on the base plate, and thus to move the plunger engagement member 904 up and down a corresponding ^"amount. Other arrangements such as rocker arm, or other mechanical connector, could also be used to translate actuator shaft movement to movement of the engagement member without departing from the present invention. Of course, the actuators for the other control functions also may be indirectly connected to the operating element as may be needed or desirable for space or operating concerns, and the present invention is not, in its broader aspects, limited to the detailed connection arrangements that may be employed for the electro-mechanical actuators .

[000271] Figures 41 and 42 illustrate loading of a disposable fluid assembly 292 into the loading station 294 of the controller 290. As seen in Figure 41, the fluid circuit assembly, as in the prior embodiment, employs a generally rigid housing or frame 308 which holds various parts of the fluid circuit assembly in a prearranged position for cooperation with various actuators within the controller 290. As seen in Figure 41, the housing or frame of the disposable fluid assembly is mounted by inserting the frame at an angle into the loading station 294, and then rotating or pivoting it into position within the loading station as shown in Figure 42. The loading station preferably has a transparent door 310 that allows the operator to view and visually verify operation of the disposable fluid circuit assembly when the controller is operating. As with the earlier embodiment, the controller includes an electro-mechanical pump actuator assembly, a vial actuator assembly, an automated frangible break assembly and a valve actuator assembly responsive to the programmed control system of the control module to control fluid flow through the fluid circuit assembly in a predetermined sequence. [000272] Figure 43 is a general plan view of the disposable fluid circuit assembly 292 employed with the controller 290 -Ill-

discussed above. As shown in Figure 43, the disposable fluid circuit assembly 292 includes a rigid housing or frame 308 that contains various parts and pieces of the disposable fluid circuit assembly in a manner analogous to the rigid tubing organizer 46 described earlier. The disposable fluid circuit assembly 292 also includes a container 312 of the reconstitution liquid, a container 314 of red blood cells, or other biological fluid to be treated (which is attached at the treatment site) , and a combination incubation and adsorption container 316.

[000273] The rigid housing or frame 308 is shown in more detail in Figures 44 and 45. As can be seen there, the rigid housing 308 comprises an open framework, allowing visual inspection of various components of the fluid circuit assembly held by or contained within the housing 308.

[000274] Turning to Figure 45, shown there is an exploded view of the rigid housing or frame 308 which, as can be seen there, is made up of two halves or shells 318 that capture most of the disposable fluid circuit assembly and a tubing organizer 320 therebetween, similar to the embodiment described earlier.

[000275] As may be seen in Figures 43-44 and 47, the disposable fluid circuit includes reconstitution fluid inlet tubing 322, red blood cell inlet tubing 324 and treated red cell exit tubing 326. The reconstitution fluid inlet tubing 322 extends from the reconstitution fluid container 312 to a frangible flow control member 328 and from the frangible flow control member to a piston cylinder or syringe-type pump 330. The fluid circuit also includes a treating agent vial and access assembly 332 and an optional quenching agent vial and access assembly 334, the details of which will be discussed later. Each vial access assembly includes a hollow piercing member or spike for accessing the contents of the vial, the piercing member communicating through respective tubing segment 332t and 334t to a connector in the reconstitution tubing flow line 322.

[000276] The red blood cell inlet tubing 324 also contains a frangible flow control member 336. From the frangible control member, the red blood cell tubing 324 extends to a rigid manifold, generally at 338, which contains preformed flow paths for red cells and for reconstituted treating agent (and optional quenching agent) . In this embodiment, as in the previously discussed embodiment, treated red cells flow through a pair of serially arranged static mixers 340 which employ a series of blade or auger surfaces to repeatedly divide and rotate the combined red cell and treating agent fluid stream to provide a high degree of mixing. Also, as with the prior embodiment, the fluid circuit and organizer, as shown in Figure 47, has an over pouch or overwrap 842 that is sealed around in peripheral edges, (along the dashed line in Figure 47) to provide double containment of the fluid flow components mounted on the organizer. As with the prior embodiment, the overpouch may comprise a pair of facing flexible plastic sheets peripherally sealed together along the dashed lines to fully enclose the tubing organizer 320 and the associated tubing, connectors and fluid flow manifold mounted thereon.

[000277] The tubing organizer 320, as best seen in Figure 46, comprises a generally rigid, flat plastic plate with a plurality of upstanding walls that serve to locate and hold various parts and pieces of the fluid circuit 292 in specific preselected positions. For example, as best seen in Figures 46 and 47, the tubing organizer 320 includes upstanding walls 344 for receiving and locating tubing segments of the reconstitution fluid inflow tubing 322 and the treating agent and quenching agent tubing segments 332t and 334t. The portion of the tubing segments captured between the walls 344 may comprise a substantially rigid plastic tubing connectors 332c and 334c, which is snap-fit between upstanding walls 346 that have facing end hooks that define a catch for receiving and holding the tubing segment in place on the tubing organizer. Similarly, upstanding walls 348 of tubing organizer define passageways for receiving fluid circuit tubing and include facing hooks or detents that serve to hold the tubing in place between the walls. Similarly, the tubing organizer 320 includes upstanding walls 350 terminating in hooked ends for holding the rigid manifold 338 in place on the tubing organizer. With this construction, fluid circuit 292 may be mounted at specific predetermined locations on the fluid organizer and held securely in those locations for subsequent assembly operations, such as attachment of the overpouch 342 and assembly within the rigid housing or frame 308.

[000278] The pinch valve arrangement for the fluid circuit assembly in this alternative embodiment operates similarly to that described earlier. Tubing organizer 320 includes a valving surface 352 that extends longitudinally within slot 354 in tubing organizer. As seen in Figure 47, the valving surface extends at 90° to tubing segments through which fluid flows from the reconstitution fluid container, the treating agent vial the quenching agent vial, and the syringe type pump. Plungers (not shown in Figure 47) associated with electro-mechanical actuators on the controller 290 may be actuated as desired, to pinch one or more of the tubing segments against the valving surface, to close the tubing and prevent flow therethrough. Release of the tubing opens the tubing to fluid flow.

[000279] Turning now to a more detailed discussion of the rigid manifold 338, Figures 51-55 show the manifold and the internal fluid passageway therewithin. The manifold 338 comprises a rigid body 356 having upstanding walls 358 defining fluid passageways and chambers within the manifold. The body is sealed by a rigid plastic cover or lid 360 which is attached to the body and sealed to the outstanding walls to form the closed passageways and chambers within the manifold as may be seen in Figures 54 and 55.

[000280] Red blood cells flow into the manifold at inlet 362. The red blood cells then flow into an enlarged chamber 364 that forms a damping chamber to remove or reduce pulsations in the flow of the red blood cells as they join with the pathogen inactivation agent or other treating agent. From the chamber 364, the red blood cells flow through fluid passageway 366 to a mixing junction or Y-site 368 from which they continue along combined fluid passageway 370 to outlet 372. The pathogen inactivation agent (optionally including a quenching agent) , or other treating agent, enters the manifold at inlet 374 and flows directly to the mixing Y or junction 368 where it is combined with the red blood cell flow stream. [000281] From the outlet 372, the combined red cell and treating agent fluid streams flow through one of the static mixers 340 (as best seen in Figure 47) . The exit of the static mixer 340 is connected to inlet 376 of the manifold, which enters manifold chamber 378. The chamber 378 forms an axial mixer that mixes the combined fluid stream in additional directions so as to reshuffle the fluid stream and reduce the possibility of any unmixed red cells that may have passed through the first static mixer. Chamber 378 communicates, through aggregate or clot filter screen 380, with outlet 382 of the manifold into a second static mixer 832, through which the combined fluid streams again undergo extensive and repeated division, twisting and division, as previously described in the prior embodiment, to further add assurance that the combined fluid streams are thoroughly mixed. From the second static mixer 340, the combined fluid streams flow through tubing 326 to the combined incubation and adsorption container 316.

[000282] Turning now to the vial and the vial access assembly shown in Figures 48-50, the disposable fluid assembly, in its preferred embodiment, employs two vial and access assemblies of identical construction - one vial and access assembly for the treating agent, such as a pathogen inactivation agent, and one vial and access assembly for a quenching agent, if desired. As shown in Figures 48 and 50, the vial and access assembly 332 for the treating agent is shown in an actuated position after the vial have been depressed by a plunger of a vial actuator mechanism located on the reusable controller 294. The vial and access assembly 334 for the quenching agent is shown in the position before actuation, and before access to the contents of the vial.

[000283] Figures 49 and 50 show, respectively, exploded views of the vial and access assemblies and cross-sectional views of the vial and access assembly. Turning first to Figure 49, the vial and access assemblies have a common base 384. The base 384 has a pair of vial and access assembly mounting rings or bodies 386, into each of which is inserted a spike or piercing pin member 388 and an intermediate sleeve 390 that located between the spike and the mounting ring (as best seen in Figure 50) .

[000284] Referring to Figure 50, vial 392 is contained in a generally cylindrical vial container 394 in an inverted position, with the vial septum 396 facing the piercing end of spike or piercing pin 388. The vial container 394 has an end cap 396 that includes an outer generally cylindrical wall 398 and an inner return wall 400. The inner and outer wall of the end cap form a projecting structure which may be inserted into the mounting ring 386, when the vial container is pushed toward the base, such as by an actuator on the controller or control module 290. O-ring 404, is located between spike 388 and the inside surface of the return wall 402 to provide sealing between them while allowing relative axial movement of the piercing member within the return wall 402. The end of each mounting ring 386 includes an inward hook surface 406 which is received within an annular groove 408 in the outer wall 398 of the vial cap 396, when in the assembled position, but prior to insertion of the piercing member into vial, as best seen in Figure 50 (left side) . The outer wall of the vial cap also includes an annular groove 410 in proximity to the vial. The hook 408 engages in the annular groove 410 when the spike is fully inserted into the vial in order hold the vial and spike together in the access position (Figure 50, right side) .

[000285] Turning now to Figures 56-60, showing the piston- cylinder or syringe type pump 330 employed in this embodiment, Figure 56 is a generally side view of the piston-cylinder pump. As can be seen there, the pump includes a generally cylindrical barrel 412 having an end port 414 at one end and an end flange 416 at the other end. The barrel also includes an intermediate annular flange 418 which cooperates with the rigid frame or housing 308. The syringe pump 330 also includes a plunger 420 and an expandable sleeve 422 that extends between the open end of barrel 412 and the exterior end of the plunger to provide containment between the open end of the barrel 412 and the plunger 420.

[000286] Figures 58-60 illustrate the syringe pump within the plunger shown in various positions within the barrel. As may be seen there, the plunger 420 extends through the open end of the syringe barrel and terminates in a piston 424 of resilient material, such as latex, neoprene or other suitable material that slides in sealed contact with the inside surface of the barrel 412. The extendable sleeve 422 terminates at each end with an axially extending sleeve 426. At one end, the sleeve is sealed to the plunger and at the other end the sleeve is sealed to a cap 428 attached to the open end of the syringe barrel 412. The cap may include an upwardly extending internal guide or sleeve 430 for receiving and guiding the plunger 420. The guide may include one or more annular detents that cooperate with one or more annular slots provided in the plunger (or vice versa) to indicate selected positions of the plunger, such as fully displaced or fully extended.

[000287] Finally, referring to the treated red cell container 316, as shown in Figure 43, as noted earlier, the container 316 is a combined incubation and adsorption container. Preferably the container comprises at least two compartments, a red cell incubation compartment 432 and an adsorption device compartment 434, which are separated by a frangible seal line 436.

[000288] The incubation compartment 432 receives the treated red cells directly from the exit tubing 326. Treated red cells reside in the compartment 432 during the incubation, in the same manner as was discussed earlier in connection with systems employing a single or dedicated incubation container. After the incubation is complete, the frangible seal line 436 is opened, and the treated, incubated red cells are allowed to flow into and contact the adsorption media 438 that is contained in compartment 434. A variety of structures may be used to provide the frangible seal line 436. For example, one or more frangible flow control devices may be extend across the seal line 436 and, when broken, allow flow of red cells from the compartment 432 into compartment 434. Alternatively, the seal line 436 may be separable or peelable, allowing the walls of the container to actually separate along the seal line, fully opening compartment 434 to red cell access from compartment 432. Other means may also be provided for bringing the two compartments into communication without departing from the present invention. For example, in a still further embodiment the flexible plastic walls of the container may be sealed together along seal line 436, and a mesh material may be located between the container walls and along and within the seal line. The mesh material facilitates separation of the container walls, allowing them to effectively peel apart at the seal line when pulled apart or when pressure is otherwise applied to^' separate the container walls. Of course, entirely separate incubation and adsorption containers also may be provided if desired.

ADDITIONAL ALTERNATIVE PROCESSING SYSTEMS [000289] Figures 61-63 show an alternative rigid tubing organizer housing 440 and associated disposable fluid circuit 442 therein, and a slot housing and associated actuator assemblies that may be employed in a reusable controller to control fluid flow through two fluid circuits simultaneously. As in the earlier embodiments, the rigid organizer housing has facing shell halves 446 that capture portions of the disposable fluid processing circuit therebetween. [000290] Figure 62 best illustrates the fluid flow components located in the rigid housing. As seen there, the fluid circuit comprises reconstitution fluid inflow tubing 448, red cell inflow tubing 450 and treated red cell outflow tubing 452. The reconstitution inflow tubing is connected, via tubing segments, to a syringe-type pump 454, pathogen inactivation agent vial access assembly 456 and quenching agent vial access assembly 458. The red cell tubing and flow tubing for reconstituted inactivation agent are connected to a flow block 460 having a performed fluid path therein. The red cells first enter an enlarged accumulation or damping chamber 462, from which they flow to a mixing junction 464 where they join the reconstituted inactivation agent (and optional quenching agent) . From the junction 464, the combined fluid stream flows through serially arranged static mixers 466, 468 and 470 shown in greater detail in Figure 70, and discussed below.

[000291] Figure 63 illustrates a slot housing 472 and associated actuator assemblies that may be used in a controller to control flow simultaneously through two fluid assemblies, such as but not limited to the type of assembly shown in Figures 61 and 62. The slot housing 472 comprises two separate, parallel receiving slots 474 for insertion of a rigid organizer housing. Pairs of valve actuator assemblies 476, pump actuator assemblies 478, vial actuator assemblies 480 and frangible breaker assemblies 482, which are substantially like those described earlier, are mounted on the housing for separate control of fluid through each fluid circuit assembly.

[000292] Figure 61 shows a further alternative fluid flow system and circuit, generally designated 484, is shown there which is particularly, but not exclusively, suited for use in inactivating pathogens in a biological fluid in a closed, sterile and disposable assembly. Generally speaking, the fluid flow circuit or assembly shown in Figure 61 includes a first fluid source 486, a second fluid source 488, a static mixing portion generally designated 490, and an output receptacle or container 492 for receiving the fluids after they have been mixed. In the illustrated embodiment, the first fluid source 486 comprises a container or bag of biological fluid, such as concentrated red blood cells, which is connected to a flow junction 494 by a first fluid flow path that includes tubing 496, and an accumulation or damping chamber 498.

[000293] The second fluid source 488 in the illustrated embodiment is a source of pathogen inactivation agent such as described above, and is illustrated as a syringe pump to provide a precise flow rate, which is connected to the flow junction 494 through a second fluid flow path comprising a tubing 500.

[000294] The flow junction 494 comprises a standard Y-site or V-site as commonly used in medical fluid flow systems, although other arrangements could also be employed for joining the fluid streams such as a T-site or the like. Generally, it is preferred to bring the fluid streams together in the same general direction of flow to provide the least potential adverse effect on the red blood cells. It may be desired to inject the inactivation agent into the center of the red cell stream because the greater density red cells tend to flow more in the middle of the fluid stream, with the lighter plasma being on the outside margins of the stream.

[000295] It is contemplated that flow rate of the first and second fluids to the flow junctions will be carefully metered, particularly in a pathogen inactivation application. Accordingly, one or more of the fluid pathways may include a tubing segment, such as a length of silicone tubing if desired, for cooperating with a peristaltic pump of the type commonly used in controlling fluid flow through medical fluid circuit assemblies. Because peristaltic pumps operate by progressively compressing the tubing with a series rollers or fingers, the flow rate within the first fluid flow path varies or pulses slightly in response to the action of the peristaltic pump. To eliminate such variation in the flow rate and to allow more precise flow rate control, the first fluid pathway may include the accumulation or damping chamber 498. This chamber accumulates a volume of fluid, with a quantity of air above the fluid that acts as a cushion or spring to dampen the pulsation so that a continuous, non- pulsing or reduced-pulsing flow of fluid exits the chamber. [000296] The flow rate of the pathogen inactivation chemical may be controlled by a peristaltic pump or by an actuator that progressively depresses the plunger of a syringe pump in a programmed and controllable fashion to accurately achieve the desired linear flow rate of pathogen inactivation fluid for mixing with the red blood cells flowing through the first fluid path. For example, in a process of inactivating pathogens in red blood cell concentrate, the RBCs may be controlled at a flow rate of about 100 cc/min. and the pathogen inactivation agent at a rate of about 6cc/min. or other desired rate for good mixing results and for minimizing damage to the red ^"cells. Although a syringe pump could also be used to control the flow of red blood cells, peristaltic pump control of red blood cells directly from their storage or collection container may be more gentle and result in less hemolysis .

[000297] As pointed out earlier, it may be important in a variety of applications, not limited to treatment of biological solutions or pathogen inactivation treatments of blood or blood components, to obtain a very high degree of mixing of two or more fluid streams and to provide a very high degree of homogeneity in the resulting mixture. In accordance with the present invention, the fluid circuit 484 shown in Figure 61 includes a static mixing portion or area 490 to achieve thorough mixing of the pathogen inactivation fluid with the concentrated red blood cells to provide a very high degree of assurance that suitable contact is made between the pathogen inactivation agent and the concentrated red blood cell fluid. In the illustrated embodiment, the static mixing area 490 includes a first mixing section 502, a second mixing section 504 and a third mixing section 506. While this may be the preferred number and order of the mixing sections, it is in keeping with the present invention to use the first mixing section alone, if it is so desired, to provide a very high degree of homogeneity in the resultant mixture. It is also in keeping with the present invention to employ the first mixing section in combination with a second mixing section to a greater degree of confidence that the sufficient mixing has occurred.

[000298] The static mixer employed in the first mixing section of Figure 61 is best seen in Figure 65. As shown in that figure, which illustrates a portion of the first static mixing section, the static mixing section includes at least one and preferably a plurality of mixers or mixing elements 508 to mix the fluid stream as it flows along the combined fluid flow path, without the need for any moving parts or external forces, other than the force of the moving stream itself. Each mixing element 508 preferably comprises a generally planar surface that is shaped to divide the fluid stream and to rotate the fluid stream as it passes by the mixing element as described earlier in connection with Figure 24.

[000299] The alternative clockwise-counterclockwise arrangement of mixing elements 508 described above in the first mixing section produces rapid mixing and creates a level of turbulence in the fluid stream which further encourages mixing as the fluid streams pass through the first mixing section. Alternatively, the mixing elements may be arranged to turn or twist the fluid in the same direction to provide a more laminar mixing flow with less of a pressure drop through the mixing section. Such an arrangement is illustrated in Figure 66, which shows a portion of a mixing section having three mixing elements, each mixing element having the same shape so as to rotate or twist the fluid stream in the same direction as described in connection with Figure 25. [000300] The arrangement and shape of the mixing elements 508 in the first static mixing section 502 results in a mixing of the fluid streams mainly in a radial direction. It may be possible in certain unpredictable circumstances for minute fluid streams to pass through the first mixing section without being mixed sufficiently to provide the highest levels of homogeneity that may be desired, for example, when mixing a pathogen inactivation agent with biological solutions such as blood or blood components. Accordingly, the combined fluid flow stream may include a second mixing section 504 which has a mixer adapted to mix the fluid in a direction other than radial, such as axially relative to the direction of flow through the first section. Such a mixer may be of a variety of shapes or configurations. As shown in Figure 64, for example, the mixer in the second section is a generally enlarged accumulation chamber or reservoir in which the combined fluids flow and disperse in directions other than radial. Any unmixed fluid streams from the first mixing section are broken up and redistributed within the second mixing section. Additional and other static mixer alternatives for use in a second mixing section will be discussed later.

[000301] Although the mixing section could terminate at the end of either the first mixing section or the second mixing section, as indicated above, to provide an even greater degree of confidence with respect to the homogeneity of the resulting mixture, and to further process any unmixed streams that are broken up in the second mixing section, a third mixing section 500 may be provided as shown in Figure 61.

[000302] From the third mixing section, the mixture flows preferably into the output receptacle or container 492, where it may be stored until needed, further processed, and/or transferred to an optional transfer container which may be preattached or attached by a later sterile connection (if employed in a medical application) .

[000303] Figure 67 shows a further alternative of the present invention, employing a recirculation feature or step to provide additional mixing. As shown there, a container of red blood cells 510 is connected via tubing 512 to a flow junction 514, where a pathogen inactivation agent is added from a pathogen inactivation fluid source 516. The combined fluid stream exits the junction and flows into a mixing section 518 which may comprise a plurality of mixing elements substantially as described in the first mixing section of Figure 64. From this mixing section, the combined fluid stream is directed into an output bag or container 520, which may also serve as a second mixer. The output bag includes a draw tube 522 which extends into the bag and near the lower end of the bag, to draw the combined red blood cell concentrate and pathogen inactivation fluid from the bag and recirculate it into the red blood cell line leading from the red blood cell container 510 to flow junction 514. From junction 514, the combined red blood cell and pathogen inactivation agent drawn from the bag are recirculated through the mixing section 518 and back into the output bag 520. This cycle of draw and mixing may be completed as many times as desired to provide the desired assurance of sufficient mixing of the red blood cell concentrate and pathogen inactivation fluid. In this embodiment it is contemplated that the red blood cell flow is controlled by a peristaltic pump and a tubing segment 524, such as a length of silicone tubing, is provided for cooperation with the peristaltic pump. The pathogen inactivation fluid source may be syringe type pump, or a peristaltic pump may also be used to control the inactivation agent flow rate.

[000304] Figure 68 shows a further embodiment of the present invention combining various features of Figure 64 and Figure 67. As in Figure 67, this system includes a red blood cell concentrate container 526 which is connected via tubing 528 to flow junction 530, where it is combined with pathogen inactivation fluid flowing from the pathogen inactivation fluid source 532, such as a syringe pump or other source. From the junction 530, the combined fluid stream is directed to a first mixing section 534 which is preferably constructed as described in connection with the first mixing section in Figure 64 -- employing a plurality of mixing elements 508 to repeatedly divide and alternately turn or twist the combined fluid streams in opposite directions as it passes therethrough.

[000305] After passing through the first mixing section 534, the fluid stream enters a second mixing section 536, which is in the form of a larger diameter tubing segment that operates comparably to an accumulation reservoir in that it allows mixing of the combined fluid streams in a direction other than radial to break up any unmixed streams and to further assure that mixing has taken place in various directions. From the second mixing section, the combined fluid stream is passed into a third mixing section 538 comprising a plurality of one or more mixing elements 508, preferably essentially identical to that described in connection with the first mixer in Figure 64. The combined fluid stream exiting the third mixing section 538 is directed into an output container 540. [000306] The output container may include a draw tube or, alternatively, the walls of the container may be vertically sealed at 542, stopping short of the bottom of the container, to create a draw area 544 that extends into the bottom of the container and through which the container contents may be drawn for recirculation through recirculation line 546 into a fluid junction 548 in the red blood cell concentrate tubing. The red blood cells and pathogen inactivation agent mixture may be recirculated through the mixing sections and output

I container as many times as desired to assure confidence that the appropriate mixing has occurred. Accordingly, this system combines three mixing sections, such as seen in Figure 64, with the output container and recirculation feature illustrated in Figure 67 to provide a system that can provide an even higher level of confidence in the completed mixture homogeneity.

[000307] Figure 69 shows yet a further alternative embodiment of the present invention in which a container of concentrated red blood cells 550 is attached by tubing 552, via frangible flow control member 568, to an accumulation or damping chamber 554 and from there to a flow junction 556 where pathogen inactivation fluid is added from a pathogen inactivation fluid source 558 such as a syringe pump or other source. From the fluid junction, the combined fluid flows serially through three separate mixing sections 560, 562 and 564, each of which is constructed essentially as described for the first mixing section of the fluid flow system shown in Figure 64. From the third mixing section, the combined fluid stream is conducted into the output container 566.

[000308] Figure 70 shows yet a further embodiment of the present invention, in which the mixing system may be part of a flow control unit or module and employing an alternative design of a second static mixing section for mixing in a direction other than radial. In this embodiment red blood cells are received, through fluid flow line 570 into an accumulation or damping chamber 572. A pathogen inactivation agent is received from a pathogen inactivation fluid source through flow path 574 to junction 576, where it joins the concentrated red cell flow. From the junction, the combined fluid streams pass through a mixing portion or area, generally at 578, having three separate mixing sections. The first and third mixing sections 580 and 584 each have one and preferably a plurality of mixing elements 508 such as described in connection with the first mixing section of Figure 64, where the combined fluid stream is repeatedly (preferably more than ten times) divided and turned or twisted alternately in clockwise and counterclockwise directions.

[000309] A second mixing section 582 is provided between the first and third mixing sections for mixing the fluid in a direction other than radial. In this embodiment, the second mixing section comprises an elongated flow path 586 with flow obstructers 588 located along the flow path. These flow obstructers extend across the flow path and require the fluid flow stream to divide and flow around the flow obstructers. This generates areas of vortices behind or downstream of each flow obstructer, as seen in Figure 71. As a result, substantial mixing in a non-radial direction, such as axial or other direction relative to the direction of fluid flow, occurs as the fluid passes through this second mixing section. As a result, the combined fluid stream that exits the third mixing section is better assured of having a high degree of mixture homogeneity.

[000310] Figures 74 and 75 show other embodiments of the second mixing section of the static mixer of Figure 70. In these embodiments, in addition to the flow obstructers 588, the walls of the flow channel may have lateral projections 590 that define restricted flow path regions through the second mixing section. The flow rate of the fluid stream increases in these restricted regions, as compared to the flow rate in unrestricted regions, and one result of this arrangement is the creation of lower pressure vortex generating areas downstream of each projection that tend to provide additional mixing in directions not limited to radial directions to aid in reshuffling the components of the fluid stream and breaking up any unmixed regions of the two fluids .

[000311] A variety of arrangements of flow obstructers and/or projections may be used in the second mixing section. In Figure 75, for example, projections 590 extend from the opposite side wall of the second mixing section. The projections are located intermediate the flow obstructers. Alternatively, the projections could alternatively be located on opposite walls with one projection projecting from one wall and the next sequential projection projecting from the opposite wall. Also, the projections may be used by themselves, without the flow obstructers.

[000312] When the apparatus and method of the present invention are used for mixing an agent, such as a pathogen inactivation agent, with a biological fluid comprising cellular material, such as red blood cell concentrate, it may be desirable to employ a premix chamber between the flow junction and the first static mixer. The premix chamber may be simple chamber where the fluids are allowed to mingle. [000313] Because the flow stream of biological material may have a density gradient, with the cellular matter or cell population centrally located and the suspension liquid located more radially outwardly, uniform mixing at the junction may not take place. By placing a premix chamber in the flow stream immediately downstream of the mixing junction, additional mixing is encouraged before entry into the mixing section. The premix chamber acts as a diffuser, in effect a type of static mixer, to help diffuse the agent and biological fluid streams and break up concentrations of either fluid. [000314] Figure 72 shows apparatus that may be used in the biological fluid stream or in the combined fluid streams to reduce any density gradient that may occur in the stream. As discussed earlier, certain flow streams, such as a flow stream of concentrated red blood cells or other cellular material may develop a density gradient where the higher density materials tend to flow in the center of the stream. The apparatus of Figure 72 addresses this by reversing this flow relationship and thereby conditioning the fluid stream for further processing.

[000315] The apparatus of Figure 72 includes an upstream flow divider 592 and a downstream flow combiner 594. A flow stream which may have a density gradient enters the upstream flow divider in the direction indicated. The flow divider has a central flow orifice 596 that diverts the inner or generally central portion of the flow stream into bypass tubing 598 and an annular flow orifice 600 that diverts the radially outer portion of the flow stream into bypass tubing 602. The flow combiner 594 introduces the fluid from bypass tubing 602 into the center of the flow path and the fluid from the bypass tubing 598 into an outer annular region of the flow path, thereby reversing the relative positions of those portions of the fluid stream to reduce the density gradient. With this reversal, the fluid stream is better conditioned for contact with a treatment agent or for entry into downstream static mixers, to assure more complete contact between the fluids. [000316] Figure 73 shows yet another style of static mixer 604 that may be used to cause axial or non-radial mixing of a fluid stream or be used as a premix chamber. This static mixer includes an inlet region 606, and diametrically enlarged diffusion region 608 and a tangentially located outlet port 610. Fluid enters the inlet region, accumulates and diffuses in the diffusion region, and exits tangentially through the outlet port 610. The result is a substantial diffusion and mixing of the fluid stream in non-radial directions. [000317] Figures 76 shows a further alternative with first and third mixing sections 612 and 616 as described above in connection Figure 11. The second mixing section 614 comprises an accumulation chamber or reservoir 618 that tends to cause mixing in a variety of directions and reshuffles or breaks up any unmixed regions of the fluids.

MIXING ALTERNATIVES [000318] The embodiments discussed above achieve mixing by manual massage or shaking of a mixing container (Figure 2) , or one or more static mixers to assure complete mixing of the red cells and treating agent (e.g., Figure 6) . Other mixing arrangements may also be used without departing from the present invention, and Figures 77-78 illustrate two further examples of alternatives mixing systems that may be employed. [000319] In Figure 77, the mixed red cells and treating agent are directed via tubing 620 into a flexible container 622, which is preferably a flexible bag. The bag is placed in a mixer 624. The mixer has divider 626 that is movable so as to press the walls of the container together at a central location to divide the container into two or more compartments (see Figure 51) . The divider 626 extends only partially across the container, leaving an open passageway connecting the bag compartments. Pressure members 628 overly each bag compartment. By alternately and oppositely compressing and releasing each compartment, the combined red cells and treating agent are gently forced back and forth between the compartments mixing them without causing unacceptable levels of hemolysis.

[000320] The compartments may be compressed automatically via electromechanical actuators, rotating cams or any other suitable means to cause continuous, preferably slow reciprocation of the actuators to move the combined red cells and treating agent gently back and forth between the compartments .

[000321] This gentle mixing may be continued on for sufficient period to achieve the desired mixing and then the container 352 may be removed from the apparatus and incubated for the desired time, e.g., about 12 hours to allow the pathogen inactivation agent to react completely with the red cells. Alternatively, the container can remain in the mixer during incubation, with the mixer being operated at a much slower rate to gently agitate the combined red cells and pathogen inactivation agent to maintain the red cells in suspension and enhance treating reaction. Treated red cells may exit through tubing 630 to container 632 for further processing, as desired.

[000322] Figure 79 shows another mixing container/apparatus that may be employed in the treating process of the present invention. As seen in Figure 79, mixing container 630 has inlet 632, outlet 634 and two chambers 636 and 638 connected in series by a passageway 640.

[000323] The mixing container 630 may be mounted in apparatus having reciprocating pressure members or paddles 642. The apparatus may have a fixed platen or support surface on one side of the container, with movable pressure members 642 positioned to compress the chambers against the platen. Alternatively, the apparatus may include opposed movable pressure members .

[000324] Whatever the construction, the apparatus preferably operates by alternately compressing each chamber to express the red cells and treating agent from one chamber, e.g., 636, through passageway 640 and into the other chamber, e.g., 638. This action is carried out repeatedly to move the combined red cells and treating agent (and optional quenching agent) gently back and forth between the chambers to thoroughly mix the red cells and treating agent. The red cells and treating agent may be transferred back and forth as many times as desired (e.g. 100 or more) to assure the desired mixing. When the mixing is completed, the combined red cells and treating agent are preferably expressed into an adsorption container for removal of any remaining treating agent, quenching agent or reaction by-products .

ALTERNATIVE CONNECTORS [000325] Although the treatment system described above employs frangible flow control members that are opened by bending, other frangible flow control members or sterile connectors may also be used. Two different versions of such connectors are illustrated in Figures 85-90.

[000326] The first version, seen in Figures 85-87 employs a base 644 that mounts a hollow spike or piercing member 646 and a fluid flow port 648, and a cap 650 with a fluid flow port 652. The spike 646 is received within a closed-end tube 654. An insert member 656, having an interior diaphragm 658, is located between the cap 650 and the closed end of tube 654. A collapsible sleeve or bellows 658 extends between the base 644 and cap 650, and encloses the spike 646, tube 650 and insert member 656.

[000327] When fully assembled, but prior to actuation, the connector appears generally as shown in Figure 54, with fluid flow tubing (not shown) connected to ports 648 and 652 and leading to the rest of the fluid circuit assembly. The connector may be located in the red cell inlet flow path, the reconstitution fluid path or other location as desired. Preferably the connector is st.erilized, such as by radiation, and the sleeve 658 protects the interior from contamination. [000328] Until actuation, the connector blocks flow through the respective fluid path in which it is mounted. To open the connector to fluid flow, the base 644 and cap 650 are pushed toward one another, forcing the piercing member 646 through the closed end of tube 654 and through diaphragm 658. [000329] Figures 88-90 show an alternative flow control member or device including a base 660, having a hollow piercing member or spike 662 and a port 664, and a cap 666, having an annular skirt 668 and a port 670. The piercing member 662 is sealingly received within one end of a tapered sleeve 672, and an insert 674 is sealingly received within the other end of sleeve 672. The insert is sealingly attached over a raised interior ring 676 in the cap, and has an interior diaphragm 678 so that when the flow control member is assembled as shown in Figure 89, the piercing member is sealed within the tapered sleeve 672 and insert 674.

[000330] As with the earlier described flow control member, the device shown in Figures 88-90 is preferably sterilized after assembly. Before actuation, as seen in the position shown in Figure 89, the flow control device blocks flow therethrough. To open the device to fluid flow, the base and cap are pushed axially together to force the piercing member 662 through diaphragm 678, providing a flow path between the ports 664 and 670. The tapered sleeve 672 accommodates this motion by bending or flexing as seen in Figure 90, and may include a weakened or annular thin wall region intermediate the ends to lessen the compressive force required to open the flow control device.

[000331] Further alternative flow control members are illustrated in Figures 91-93. In Figure 91, the device includes inner and outer tubular telescoping members 680 and 682. Inner tube 680 has a tubing connection port 684 at one end and an open opposite end that is sealed by an overcap 686. The overcap is frangible and in the illustrated embodiment includes a line of reduced thickness 688 defining a weakened area.

[000332] The outer telescoping member 682 has an outer sleeve 690 and an inner tapered breaking member 692 for engaging and breaking the frangible overcap of the inner tube to open flow through the device. The inner and outer telescoping members are relatively slidable from the telescoped-apart position shown in Figure 91, where flow is blocked by the overcap, to a telescoped-together position where the projecting breaking member 688 has pierced the overcap opening flow through the flow control device. 0-ring 694 between the inner and outer members allows a relative sliding motion between the inner and outer members while sealing against contaminants. [000333] The flow control device of Figure 92 is similar to that described immediately above, with a hollow inner telescopy member 696 and a hollow outer telescoping member 698. The outer telescoping member 698 has an inner projecting wall 700, the terminal end of which is employed to puncture a frangible overcap 702 that covers the open end of inner telescoping member 696. When it is desired to open the flow path, the inner and outer members are pushed together, either manually or automatically by actuation device, forcing the projecting wall 496 through the overcap. This opens the connector to flow therethrough between ports 704 and 706. [000334] Figure 93 illustrates another form of flow control device that could be employed in the illustrated method and apparatus. As shown there, the device includes a tubing receiver 708 in the form of a u-shaped rigid plastic member with a raised central occluder 710. Each side of the receiver is open to rotably receive a hub or axle 712 of a cam member 714. The cam member has an off-center, non-circular (such as hexagonal) bore 716 to receive a drive shaft 718 of matching cross-sectional shape.

[000335] The drive shaft may be associated with the controller and be selectively rotatable by an electromechanical actuator. The cam member is located on flexible tubing 720 associated with a disposable fluid assembly. The drive shaft 718 rotates the eccentric cam 714 between a position where it compresses or pinches the tubing 720 against the occluder 710, blocking flow through the tubing, and a position where it releases the tubing to allow flow through the tubing. Intermediate positions may provide intermediate flow rates.

VIAL/VIAL ACCESS [000336] Alternative vial containers and vial access devices are shown in Figures 94-96. Figure 94 shows a vial 722, a vial container 724, a hollow access (piercing pin or spike) member 726 for accessing the contents of vial 722, which may contain a red cell treating or other agent. Vial 722 is situated within vial container 724, which is sealed by end cap 728 to fully enclose and provide double containment of the vial .

[000337] The vial container 724 includes the integral access member 726 to access the vial contents. The access member is located within a raised ring 730 at the closed end of vial container 724. The ring has an internal annular rib 732 which supports the vial 722 above the piercing member prior to piercing.

[000338] To access the vial contents, the end cap 728 is depressed, moving the vial past the rib 732 and against the piercing member, and forcing the piercing member through the vial septum and into the interior chamber within the vial . The cap may be depressed manually or automatically by an actuator of a controller or other device.

[000339] Further, alternative structures for the vial container are shown in Figures 95-96. In Figure 95, the piercing member 726 is separate from the vial container and attached to the vial container, such as by press fit or bond. [000340] In Figure 96, the piercing member 726 and ring 730 are integral, and separate from the vial container 724. They are attached to the container by press fit, bond or other technique.

[000341] Figures 97-98 illustrate another vial/vial container access structure that may be employed in the red cell treatment system described herein. As shown there, vial 734 is sealed in vial container 736. The vial 734 is of typical construction with a glass or plastic body 738 and piercable septum or closure 740.

[000342] The vial container 736 has a container body 742, a vial container cap or cover 744 mounted on the container body, with a piercable diaphragm 746 captured between the body 742 and cap 744. The cap includes a cylindrical extension 748 for slidably receiving a hollow spike or piercing member body 750. O-ring 752 provides a seal between the spike body 750 and the interior surface of the cap extension 748 while permitting relative slidable movement. Piercing member 750 is hollow and communicates with a connection port 754 that may be attached to fluid flow tubing or other apparatus.

[000343] Figures 99-100 show yet a further embodiment of vial and vial access apparatus 756 that may be used. The apparatus there includes a spike or piercing member body 758, a cap or cover 760, an internal vial support member 762, vial 764 and optional connecting ring 766. The piercing member body 758 has an elongated cylindrical body 768 for receiving the vial 764 in an inverted position. Attachment of the cover 760, which may be by direct bonding to the open end of cylindrical body 768 or by the connecting ring 766, seals the vial within the housing to provide double containment.

[000344] The piercing member body 758 includes an axially extending hollow spike or piercing member 770 positioned to pierce the vial septum 772 when the cover 760 is depressed. [000345] Internal support member 762 supports the vial in a position spaced above the spike until access is required. The internal support member is sized to frictionally engage the inside surface of spike body 768 at a location above the end of spike 770. The inside diameter of the spike body 768 may decrease (gradually or abruptly) in the direction toward the spike so that the support member 762 can be easily inserted into the open end of spike body 768 (before the cover 760 is attached) and positioned at the desired location, at which point it is frictionally engaged against inside surface with sufficient force to reliably support the vial 764 above the spike 770 until actuation is required.

INCUBATION [000346] As discussed above, following mixing of the red cells and treating agent, the combined fluids are preferably incubated to allow the treating agent sufficient time to react with or otherwise carryout the desire treatment of the red cells. Figure 101 shows one potential incubation process employing (but not limited to) a dual container mixing arrangement similar to that shown in Figure 79 above. As illustrated in Figure 101, after mixing is completed the treated red cells are collected in an incubation container 774 that is pre-attached to the adsorption or compound adsorption device "CAD" container 776. The CAD container, which is sealed from the incubation container, is folded into a compact arrangement with the incubation container and placed in an incubation tray 778. The tray may be stored in stationary or movable racks or otherwise arranged conveniently for incubation. Preferably the trays are mounted an apparatus that provides for gentle oscillation or shaking of the tray and the bags thereon, keeping the red cells suspended and encouraging mixing and reacting with the treating agent. [000347] After incubation is completed, the flow connection between the incubation CAD and containers is opened and the treated red cells, treating agent and optional quenching or other agent, are transferred into the CAD container 776. Although illustrated as a completely separate container, the CAD container 776 may be a separate compartment in the incubation container, or the adsorption device may be separately contained and releasable into the incubation container when desired. In the illustrated embodiment employing a separate container, the filled CAD container 776 is sealed and severed from the incubation container and moved to an adsorption location, where it rests for such time as required for the adsorption device to remove any unreacted treating agent or quenching agent or reaction by-products. During this time, the container is preferably subject to gentle oscillation or shaking to enhance contact with the adsorption device and to maintain the red cells in suspension. [000348] Figure 102 illustrates a potentially more automated incubation and adsorption station employing an incubation/adsorption controller 780) using a two-container arrangement similar to that described above. In this embodiment, however, a frangible flow control member generally at 782 is located in tubing connecting the incubation and CAD containers 784 and 786.

[000349] The incubation and CAD containers are mounted in a spaced apart arrangement on the incubation/CAD controller 780. The controller includes a frangible breaker 788 for automatically opening the frangible flow control member after incubation is completed and means for automatically transferring the red cell contents of the incubation to the CAD container.

[000350] The means for transferring the red cells in the incubation container 784 may contact or squeeze the incubation container to forcibly express the contents into the CAD container 786. Alternatively, the controller may invert the containers to allow transfer by gravity drain from the incubation container 784 to the CAD container 786. However transferred, the controller may monitor the storage time in the CAD container and signal the operator when the adsorption cycle is completed. In this embodiment, the controller may also apply a gentle shaking or oscillation to the incubation and CAD containers to maintain the red cells in suspension and encourage the respective treatment .

[000351] Figure 102 also depicts a manual device 790 for transferring treated red cells from the incubation container 784 to a CAD container 786 and for sealing and severing the tubing between them.

[000352] When the incubation and adsorption steps are complete, the flow tubing between the containers is sealed and severed, any required samples are taken and the CAD (or dedicated container) are transferred to a refrigeration facility for storage. At this time, any required information may be entered into the data management system. It is further contemplated that the incubation/adsorption controller 780 would, by appropriate connection into a data management system, record the beginning and completion of each step for each quantity of blood treated and any other data that may be desired for record or archival purposes.

[000353] A further embodiment of an incubation and adsorption apparatus is shown in Figures 103, 104 and 105. As shown there, an automated incubation/adsorption controller 790 comprises a tower 792 having a plurality of slotted carriers 794 arranged vertically in the tower. The carriers are rotatable up to about 135° in each direction. The tower also includes a series of vertically - arrayed hanging locations 796 along each side of the tower.

[000354] The tower 792 is intended to operate simultaneously on a series of disposable container systems, generally at 798. Each container system includes an incubation container 800 and an adsorption or "CAD" container 802. The container is connected by flexible tubing including a frangible flow control member 804. The container system may have been part of a larger disposable fluid circuit assembly, much of which has been discarded after a reconstitution and/or mixing process has been carried out. [000355] The incubation container 800 of each container system 798 is suspended at a hanging station 796 on the tower, the frangible member 804 is mounted on an automated frangible breaker 806 in the tower, and the adsorption container is inserted into one of the slots of carrier 794. Each carrier has a pair of slots and is associated with a pair of frangible breakers 806 and hanging stations 796 located on opposite sides of the tower. This permits incubation and adsorption to be carried out simultaneously on two units or quantities of red cells. The hanging station may comprise a simple hook or include a scale to monitor the container weight. The hanging station also may include means to agitate the container to keep the red cell solution suspended.

[000356] The tower may have an associated control system to monitor the incubation period and to automatically open the frangible member after the incubation is complete. The control system may also monitor a weight scale on which the incubation bag is suspended to assure that transfer to the adsorption container has occurred, and generate a warning if the transfer to the adsorption container does not occur according to the desired protocol.

[000357] After transfer of the treated red cells to the adsorption container (and during transfer if so desired) , the apparatus agitates the adsorption container by gently rotating the carrier back and forth (clockwise and counter-clockwise) through an arc of about 270 degree, as illustrated in Figure 105. This oscillation serves to keep the red cells suspended and enhance contact with the adsorption device located in the adsorption container. When the adsorption period is completed. The apparatus may signal completion by aural or visual signals or electronically or otherwise.

DATA MANAGEMENT

[000358] Figure 106 diagrammatically shows a data management system that may be used in the red cell (or other biological fluid) treatment described above. Figure 106 depicts for illustrative purposes lines of communication between a data manager 808 and one or more receiving/preparing stations 810, mixing stations 812, incubation and adsorption stations 814 and 816 (which may be a single integrated station or separate stations) and final processing/check stations 818. Of course, these stations may combined or further divided, as convenient for processing and data management purposes. Also, there may be additional computers or servers 820 that communicate directly with the stations and control or relay the communications to the Data Manager.

[000359] The data manager 808 may be an on-site or off-site general purpose data processing system, such as a digital microprocessor-based programmable computer system having memory, logic and communication capabilities. Within the data manager resides the desired protocol or process steps or characteristics for the particular red cell or other biological fluid treatment - red cell pathogen inactivation in this particular embodiment.

[000360] The data manager 800 may be in communication with the various stations or apparatus by any suitable wire or wireless communication techniques. For example, the various stations may be in a local area network, or part of a local area network. Communications to and from the data manager may also be by way of the local area network. Alternatively, any communications between the various stations and to/from the data manager may be by the world wide web internet, other data processing systems, the public switched telephone network, any cellular network, satellite communication links, radio frequency (RF) links, Wireless Fidelity (Wi-Fi) technology, or any combination thereof. Any ^' other suitable communications technologies or combination thereof may also be employed. [000361] Each station may employ a miniature internet web server with its own unique URL, such as a PicoWeb server from Lightner Engineering of La Jolla, California. This would allow access to each station via world wide web internet connection from the data manager. One or more monitors 822 may be employed to allow remote user access to the data manager, to the individual stations and to any other computer or sever 820 for viewing data, communicating instructions or for other purposes. Of course, the data manager may be connected to other on-site or off-site computers for access by others, if so desired.

[000362] As depicted in Figure 106 each station preferably has data input means such as keyboard 824, bar code scanner or wand 826 or other device for entering data, such as from a bar code label 826 on a container of red blood cells 828 or a data-containing communications/computer chip in or on the container. Each station may also be configured and equipped to communicate directly with the data manager without user or attendant intervention. For example, start and completion of the incubation cycle for a particular quantity of red blood cells may be communicated automatically and directly to the data manger 808 or intervening computer 820 on a real time basis, or such data may be stored at the particular stations until the station is interrogated by the data manager. However the data is transmitted, the data manager is preferably programmed to compare the processing sequence and conditions to the established protocol and to provide a means to verify and certify that all the required processing steps/conditions have been met for each particular quantity or unit of red cells treated. Such certification could be, for example, by way of a label that is applied automatically or manually to the container of treated red cells at the final processing station. Other means for certifying such processing may also be used. If the required processing steps or protocol are not met, the data manager may also provide an indication that processing was incomplete or not verifiable, and may provide a decertification for that particular quantity or unit.

[000363] Although the present invention is described in connection with certain examples or embodiments employed in red cell pathogen inactivation, it is understood that the system apparatus and methods described above may also be used in treating other biological or medical fluids and is not limited to only the particular application described.

Claims

CLAIMSWhat is claimed is:

Claim 1. A method for ex vivo treatment of red blood cells comprising: providing a quantity of red blood cells; providing a disposable fluid circuit assembly and a reusable controller, the fluid circuit assembly and controller being cooperative to control fluid flow through the fluid circuit pursuant to a predetermined operation sequence including at least one treatment step to affect the red cells; introducing the red cells into the disposable fluid circuit; processing the red cells through the fluid circuit assembly in accordance with the predetermined operation sequence, including treating the red cells in accordance with the treatment step to affect the red cells; and monitoring compliance with the predetermined operation to assure that the red cells are processed through the fluid circuit in accordance with the predetermined operation sequence .

Claim 2. The method of Claim 1 in which the quantity of red cells is associated with a unique identifier and the method includes electronically storing the results of monitoring in association with the unique identifier.

Claim 3. The method of Claim 1 in which the controller includes an electronic data storage memory, and the monitoring includes automatic electronic monitoring of the predetermined operation sequence and generating a compliance report, the compliance report being stored in said data storage memory.

Claim 4. The method of Claim 1 wherein the predetermined operation sequence includes a plurality of steps, each such step including at least one monitored condition, the monitoring being carried out automatically and the results of the monitoring being retrievably electronically stored in association with a unique identifier.

Claim 5. The method of Claim 4 in which the steps include: contacting the red blood cells with a treating agent for a selected time; and contacting the combined red blood cells and treating agent with a sorption device for a selected time to remove any treating agent or reaction by-products.

Claim 6. The method of Claim 1 in which the predetermined operation sequence includes one or more treatment steps for pathogen inactivation, blood type universalizing, leukocyte reduction or inactivation or prion removal or inactivation.

Claim 7. The method of Claim 1 in which the predetermined operation sequence includes contacting the red cells with a treating agent.

Claim 8. The method of Claim 7 in which the treating agent comprises a pathogen inactivation agent, or a blood type universalizing agent.

Claim 9. The method of Claim 7 wherein the treating agent is liquid and contacting comprises mixing the treating agent with the red cells.

Claim 10. The method of Claim 7 in which the predetermined operation sequence includes removing the treating agent or reaction by-products from the red cells.

Claim 11. The method of Claim 1 including a plurality of treatment steps to affect different characteristics of the red cells .

Claim 12. The method of Claim 11 including a treatment step for pathogen inactivation and a treatment step for blood type universalizing.

Claim 13. The method of Claim 11 in which the treatment step for blood type universalizing includes contacting the red cells with one or more enzymes adapted to cleave the antigens that characterize type A, B and/or AB blood types.

Claim 14. The method of Claim 13 including removing the enzyme and reaction products or by products after the treatment steps .

Claim 15. The method of Claim 14 in which the removing includes washing the red cells .

Claim 16. The method of Claim 7 including substantially removing the treating agent by washing the red cells.

Claim 17. The method of Claim 12 which the treatment steps for pathogen inactivation and blood type universalizing are carried out at least on part simultaneously.

Claim 18. A disposable pre-assembled fluid circuit module for use with a reusable control unit for treating red cells in accordance with a predetermined operation sequence, the disposable module comprising: a fluid inlet; a red cell inlet; a fluid outlet; a sealed container having an interior chamber containing a red cell treating agent; a container access member for accessing the interior of the sealed container; a fluid pump; a flow path interconnecting the fluid inlet, red cell inlet, fluid outlet, pump and container access member.

Claim 19. The module of Claim 18 in which the sealed container comprises a vial having a piercable wall and the access member comprises a piercing member, the vial being slidably movable toward the piercing member by the control unit to access the contents of the vial.

Claim 20. The module of Claim 19 in which the fluid pump comprises a cylinder and piston movable within the cylinder, the piston being operable by the control unit to precisely control fluid flow.

Claim 21. The module of Claim 18 further comprising a static mixer in communication with the flow path for mixing the agent and the red cells.

Claim 22. The module of Claim 18 further comprising a second sealed container having any interior chamber containing a second agent .

Claim 23. The module of Claim 18 in which the agent is a dry material reconstitutable by the addition of liquid.

Claim 24. The module of Claim 22 in which the red cell treating agent is a pathogen inactivation agent and the second agent is a quenching agent .

Claim 25. The module of Claim 24 in which the inactivation agent and quenching agent are in dry form and reconstituted by the addition of fluid by the pump.

Claim 26. The module of Claim 18 in which one or both of the sealed container and access members are movable by the control unit to access the treating agent in the interior chamber .

Claim 27. The module of Claim 26 in which the sealed container comprises a vial slidably movable by the control unit and having a piercable wall, and the access member comprises a piercable member situated to pierce the wall when the vial is moved by the control unit.

Claim 28. The module of Claim 18 in which the red cell inlet communicates with a flow dampener disposed in said module .

Claim 29. The module of Claim 18 comprising a secondary containment enclosing the flow path to safeguard against leakage into the ambient environment .

Claim 30. The module of Claim 18 comprising a flow path guide for arranging the flow path in a defined manner and a rigid outer shell containing the flow path and flow path guide .

-( Claim 31. The module of Claim 18 in which said flow path includes a plurality of valve stations operatively controlled by the control unit for controlling flow through the flow path.

Claim 32. The module of Claim 30 in which the flow path includes a plurality of tubing segments and the flow path guide includes a plurality of valve areas, each valve area of the path guide comprising a rigid backstop and a tubing segment of the flow path overlying the back stop, the rigid shell including an aperture adjacent the valve area for receiving a valve element of the control unit to compress the tubing segment against the backstop.

Claim 33. The module of Claim 32 in wliich the backstop comprises a generally elongated pinch surface generally extending transverse to the axis of the respective tubing segment to pinch the tubing closed when the tubing is pressed against the backstop surface .

Claim 34. A reusable control module for controlling processing of red cells through a disposable fluid circuit module of the type including a fluid inlet, a red cell inlet, a fluid outlet, a sealed container containing a red cell treating agent; a container access member for accessing the treating agent, a fluid pump and a flow path interconnecting the above, the reusable control module including: a receiving station for receiving the fluid circuit module in operative association with the control module; a plurality of weigh stations; a pump actuator for actuating the pump on the fluid circuit module; a plurality of valves operable to control flow through the flow path in the fluid circuit module; an actuator for causing the sealed container to be accessed by the container access member; and a control system for automatically processing red cells through the fluid circuit module in accordance with a predetermined operation sequence including at least one treatment step to affect the red cells.

Claim 35. The control module of include Claim 34 further comprising a pump for controlling flow of red cells into the fluid circuit module.

Claim 36. The control module of Claim 34 for use with a fluid circuit module that includes a frangible flow control element located in the flow path, the control module further including a frangible element breaker for automatically breaking the element and opening the flow path.

Claim 37. The control module of Claim 34 for use with a fluid circuit module in which the sealed container is a vial with a piercable wall and the access member is a piercing member, in which the control module actuator for causing access to the sealed container comprises a pusher element adapted to push the vial and piercing member together until the piercing member pierces a wall of the vial for access the red cell treating agent in the vial.

Claim 38. The control module of Claim 34 for use with a fluid circuit module in which the pump comprises a cylinder and piston that are relatively slidably movable, the control module pump actuator comprising a linearly movable engagement member for moving the piston or cylinder to intake or expel fluid from the cylinder.

Claim 39. Apparatus for opening an elongated frangible flow control member of the type having a weakened area located between the opposite ends thereof, the apparatus comprising an engagement member for receiving the frangible member in proximity to the weakened area and a linear actuator operably attached to the engagement member to move the engagement member linearly against the frangible member to fracture the member at the weakened area.

Claim 40. Apparatus of Claim 39 in which the actuator is adapted to rotate the engagement member between a release position and engaged position.

Claim 41. Apparatus of Claim 40 in which the engagement member includes a recess for receiving the frangible member therein when the engagement member is in the engage position.

Claim 42. Apparatus of Claim 41 adapted to simultaneously open two parallel spaced apart frangible members, the engagement member being located between the frangible members and comprising opposed slots, the actuator adapted to rotate between a release position and an engaged position in which the frangible members are received in the opposed slots .

Claim 43. A method for opening an elongated frangible flow control member of the type having a weakened area between opposite ends thereof, comprising positioning the frangible member in a linearly movable engagement member, bracing the ends of the frangible member against linear movement and linearly

moving the engagement member to fracture the frangible member at the weakened area .

Claim 44. Apparatus for opening an elongated frangible flow control member of the type having a weakened area between the opposite ends thereof, the apparatus comprising a rotor having at least two spaced members for receiving a frangible member therebetween and a rotary actuator adapted to rotate the rotor to bend at least one end of the frangible member relative to the weakened area to fracture the member.

Claim 45. The apparatus of Claim 31 adapted to open at least two frangible connectors simultaneously.

Claim 46. A syringe pump comprising: an elongated barrel defining a generally cylindrical bore open at one end; an elongated plunger extending through the one end of the barrel, one end of the plunger disposed in and sealingly contacting the bore and the other end of the plunger being exterior of the bore; and an elongated containment sleeve having opposed ends, the sleeve being disposed around the plunger, one end of the sleeve being sealingly attached to the one end of the barrel and the other end of the sleeve being sealingly attached to the other end of the plunger to sealingly contain the plunger and provide containment against leakage from the one end of the barrel .

Claim 47. The syringe pump of Claim 46 in which the sleeve is axially extendable to accommodate reciprocation of the plunger.

Claim 48. The syringe pump of Claim 47 in which the sleeve is disposed to extend or contract in accordion-like fashion with movement of the plunger

Claim 49. A method of reconstituting a dry agent contained in a container having an internal pressure and a piercable access site, the method comprising providing a piercing member with a fluid flow path defined therein; pressurizing the fluid flow path of the piercing member to a pressure greater than the internal pressure and piercing the access site with the piercing member while maintaining the greater pressure in the piercing member flow path.

Claim 50. The method of claim 49 including flowing liquid through the piercing member into the vial container after piercing the access site.

Claim 51. The method of claim 50 in which the piercing member has a distal opening communicating with the fluid flow path and the access site has an internal surface, and piercing the access site includes positioning the distal opening generally in proximity to the interior surface .

Claim 52. The method of claim 50 in which the container has a generally longitudinal axis and the piercing member has a longitudinal axis, and piercing the access site includes positioning the piercing member such that its axis is not parallel to the axis of the container.

Claim 53. A method of reconstituting a dry agent contained in a vial having a longitudinal axis and a piercable access site, the method comprising providing a piercing member having a longitudinal axis and a fluid flow path defined therein; piercing the access site with the piercing member at an angle such that the axis of the piercing member is not parallel to the longitudinal axis of the vial.

Claim 54. The method of claim 53 including flowing liquid through the piercing member into the vial after piercing the access site, the piercing member including opposed distal fluid openings and the angle of piercing creating non-symmetrical flow of liquid from the opposed openings .

Claim 55. The method of claim 53 in which the angle is about thirty degrees .

Claim 56. A monitoring system for confirming the treatment of a selected blood product in accordance with a pathogen inactivation procedure, said monitoring system comprising: a data processing system with memory, a monitoring program stored in said memory, said monitoring program having a plurality of predetermined steps for treating the selected blood product in accordance with the pathogen inactivation procedure, and means for communicating information to the data processing system relative to selected steps of treating the blood product, whereby said data processing system compares the information communicated in each selected step of treating the blood product to the predetermined steps in the monitoring program to confirm that the blood product has undergone each of the predetermined steps.

Claim 57. The monitoring system in accordance with claim 56 wherein said data processing system also compares the information communicated in each selected step to predetermined steps to confirm that the selected steps have been executed.

Claim 58. A method of confirming the treatment of a selected blood product in accordance with a pathogen inactivation procedure in a monitoring system including a data processing system with memory, a monitoring program stored in said memory, said monitoring program having a predetermined plurality of steps for treating the selected blood product in accordance with the pathogen inactivation procedure, said method comprising: communicating information to the data processing system relating to each selected step of treating the selected blood product,

comparing the information communicated in each selected step of the treatment to confirm that the selected blood product has undergone each of the predetermined steps, and storing the information communicated in each step of the treatment of the blood product in said memory for later access .

Claim 59. A method of confirming the treatment of one of a plurality of different blood products in accordance with a pathogen inactivation procedure by a data management system, said blood product contained within a container having an identifier that identifies the type of blood product, a plurality of disposable pathogen inactivation sets with one compatible disposable pathogen inactivation set for each of the plurality of blood products, said pathogen inactivation set having identification indicia, said data management system including a data processing system with memory, a monitoring program stored in memory, said monitoring program having a predetermined sequence of steps required to treat each one of the plurality of different blood products in accordance with an appropriate pathogen inactivation procedure for each one of the plurality of blood products, and apparatus for processing the blood product during the treatment, said apparatus in communication with the data management system, said method comprising:

communicating information about the identifier on the selected blood product container to the data processing system, communicating information about the identification indicia associated with the selected pathogen inactivation set to the data processing system, attaching the container of the blood product to the disposable pathogen inactivation set carrying out the pathogen inactivation procedure, communicating data relating to the actual procedure carried out to the data processing system, whereby the data processing system compares the information communicated about the selected steps to the predetermined steps to confirm that the blood product has undergone each of the predetermined steps and to confirm that a compatible disposable pathogen inactivation set was used for the identified blood product.

Claim 60. Apparatus for mixing at least first and second fluid in a stream to provide a very high degree of homogeneity of the mixture, the apparatus comprising a mixture inlet for receiving the combined first and second fluid stream, a mixture outlet for the combined first and second fluid stream and a fluid flow path between the inlet and outlet, the flow path including a first static mixing section and a second static mixing section, the first mixing section comprising at least one radial mixer for mixing the stream in a generally radial direction relative to the direction of fluid flow through the first section and the second mixing section comprising at least one axial mixer for mixing the stream in a generally axial direction relative to the direction of flow through the first section.

Claim 61. The apparatus of Claim 60 wherein the first static mixing section comprises a plurality of flow dividers for dividing and rotating the fluid stream as it moves through the first section.

Claim 62. The apparatus of Claim 60 wherein the second static mixing section comprises a turbulence generator.

Claim 63. The apparatus of Claim 60 further comprising a third static mixing section downstream of the second static mixing section, the third static mixing section comprising a radial mixer for mixing the stream in a generally radial direction relative to the direction of fluid flow through the third section.

Claim 64. The apparatus of Claim 60 in which the apparatus is adapted to mix a biological fluid containing cellular matter and an agent fluid, and the mixers are adapted to mix the fluids without substantially harming the cellular matter.

Claim 65. The apparatus of Claim 60 in which the radial mixer is adapted to mix the stream while maintaining a generally laminar flow through the first section.

Claim 66. The apparatus of Claim 60 in which the radial mixer is adapted to create a turbulent flow through the first section.

Claim 67. The apparatus of Claim 60 in which the axial mixer comprises a fluid reservoir.

Claim 68. The apparatus of Claim 60 in which the flow path further comprises a fluid reservoir upstream of the first section.

Claim 69. The apparatus of Claim 60 in which the radial mixer comprises a plurality of surfaces, serially located in the flow path, to repeatedly divide and rotate the fluid stream as it passes through the first section.

Claim 70. The apparatus of Claim 69 in which each surface has a generally helical shape.

Claim 71. The apparatus of Claim 60 in which the axial mixer comprises a plurality of vortices-generating elements serially located in the second section of the flow path.

Claim 72. The apparatus of Claim 69 in which the surfaces rotate the fluid stream in the same direction.

Claim 73. The apparatus of Claim 71 in which each surface rotates the fluid stream in a direction opposite to that of the preceding or succeeding surface.

Claim 74. The apparatus of Claim 69 in which each surface is generally helically shaped.

Claim 75. The apparatus of Claim 71 in which the second mixing section comprises an elongated fluid flow passageway and the vortices-generating elements comprise flow obstructers located in the passageway to form a plurality of flow paths of smaller cross-sectional area around the obstructers to create vortices regions downstream of each obstructer.

Claim 76. A static mixer for mixing a fluid stream comprising a biological fluid and a pathogen inactivation agent fluid, comprising a fluid stream flow path through which the fluid stream passes and a plurality of flow dividers located within and serially along the flow path, the flow dividers being shaped to divide and rotate the fluid stream and each successive flow divider being disposed to subdivide the fluid stream as it passes along the flow path, the flow path comprising a sufficient number of dividers to provide a degree of homogeneity resulting in effective contact between the pathogen inactivation agent and the biological fluid.

Claim 77. The static mixer of Claim 76 in which the flow dividers mix the fluid stream in a generally radial direction.

Claim 78. The mixer of Claim 76 on which each flow divider rotates the fluid stream in the opposite direction to that of respective upstream or downstream flow divider.

Claim 79. The mixer of Claim 76 in which each divider is generally helical in shape.

Claim 80. The mixer of Claim 76 wherein the flow path further comprises a reservoir upstream of the flow dividers.

Claim 81. The mixer of ^l Claim 76 comprising ten or more flow dividers.

Claim 82. The mixer of Claim 76 in which each flow divider is disposed to substantially bisect the fluid stream and rotate it about 180°, and each flow divider is disposed to bisect the fluid stream at about 90° relative to the direction of bisection by the immediately upstream or downstream divider.

Claim 83. Apparatus for mixing at least first and second fluids in a stream to provide a very high degree of homogeneity of the mixture, the apparatus comprising a first fluid passageway and a second fluid passageway, the first and second fluid passageways joining in a combined fluid passageway, an accumulation chamber located in the first fluid passageway, the combined fluid passageway including a first static mixing section and a second static mixing section, the first mixing section comprising a plurality of static radial mixers for dividing the fluid stream and mixing the stream in a generally radial direction relative to the direction of fluid flow through the first section and the second mixing section comprising at least one static mixer for mixing the stream in at least a non-radial direction to provide more complete mixing.

Claim 84. The apparatus of Claim 83 wherein the first fluid pathway comprises a pumping segment upstream of the accumulation chamber for cooperation with a peristaltic pump.

Claim 85. The apparatus of Claim 83 wherein the static mixer in the second mixing section comprising an accumulation chamber.

Claim 86. The apparatus of Claim 83 further comprising a third static mixing section downstream of the second static mixing section, the third static mixing section comprising a plurality of static radial mixer for dividing the fluid stream and mixing the stream in a generally radial direction relative to the direction of fluid flow through the third section.

Claim 87. The apparatus of Claim 83 in which the apparatus is adapted to mix a biological fluid containing cellular matter and an agent fluid, and the mixers are adapted to mix the fluids without substantially harming the cellular matter.

Claim 88. The apparatus of Claim 83 in which the static radial mixer is adapted to mix the stream while maintaining a generally laminar flow through the first section.

Claim 89. The apparatus of Claim 83 in which the static radial mixer is adapted to create a turbulent flow through the first section.

Claim 90. The apparatus of Claim 83 in which the static radial mixer comprises a plurality of surfaces, serially located in the flow path, to repeatedly divide and rotate the fluid stream as it passes the first section.

Claim 91. The apparatus of Claim 90 in which each surface has a generally helical shape.

Claim 92. The apparatus of Claim 83 in which the static mixer in the second mixing section comprises a plurality of vortices-generating elements serially located in the second section of the flow path. '

Claim 93. The apparatus of Claim 90 in which the surfaces rotate the fluid stream in the same direction.

Claim 94. The apparatus of Claim 90 in which each surface rotates the fluid stream in a direction opposite to that of the preceding or succeeding surface .

Claim 95. The apparatus of Claim 92 in which the second mixing section comprises an elongated fluid flow passageway and the vortices-generating elements comprise flow obstructers located in the passageway to form a plurality of flow paths of smaller cross-sectional area around the obstructers to create vortices regions downstream of each obstructer.

Claim 96. A method for mixing two or more fluids in a combined stream comprising mixing the stream in a radial direction relative to the direction of flow of the fluid stream and mixing the fluid stream in a direction other than radial .

Claim 97. The method of Claim 96 in which one fluid is an agent adapted to act on a biological fluid and another fluid is a biological fluid.

Claim 98. The method of Claim 96 in which the radial mixing includes dividing and rotating the fluid stream a plurality of times.

Claim 99. The method of Claim 98 in which the fluid stream is alternatingly rotated in opposite directions.

Claim 100. The method of Claim 96 including mixing the fluid stream in a radial direction after mixing in a direction other than radial.

Claim 101. The method of Claim 96 in which the fluids are pre-mixed before mixing them in a radial direction.

Claim 102. The method of Claim 96 including recirculating the fluid stream and repeating the steps of Claim 37 a plurality of times.

Claim 103. The method of Claim 96 in which one fluid comprises red blood cell concentrate and another fluid comprises a pathogen inactivation agent.

Claim 104. The method of Claim 103 including separately controlling the flow rates of the one and other fluids.

Claim 105. The method of Claim 103 including conditioning the flow of the one fluid to reduce any density gradient .

Claim 106. The method of Claim 98 in which the fluid stream is divided and rotated ten or more times .

Claim 107. A method of mixing a pathogen inactivation agent fluid stream and concentrated red blood cell fluid stream comprising: joining the fluid streams to form a combined fluid stream; repeatedly dividing and rotating the combined fluid stream; mixing the combined fluid stream in a direction other than radially with respect to the direction of flow after the repeated dividing and rotating; collecting the combined fluid stream in a container.

Claim 108. The method of Claim 107 further comprising repeatedly dividing and rotating the combined fluid stream after the accumulating step and before the collecting step.

Claim 109. The method of Claim 107 including conditioning the concentrated red blood cell fluid stream to reduce any density gradient before joining the fluid streams.

Claim 110. The method of Claim 107 including flowing the fluid streams to a flow junction where they are combined and flowing the pathogen inactivation agent fluid stream to the flow junction before the red cell concentrate fluid stream is flowed to the fluid junction.

Claim 111. The method of Claim 107 including withdrawing the combined fluid from the container and repeating the steps of Claim 106.

Claim 112. The method of Claim 107 in which the mixing step comprises accumulating the combined fluid stream in a chamber .

Claim 113. The method of Claim 107 in which the mixing step comprises partially obstructing the combined fluid stream to create mixing vortices to mix the fluid in a direction other than radial .

Claim 114. A method for ex-vivo treatment of collected red cells suspected of containing pathogens comprising: combining the red cells and a first solution; combining a pathogen treatment agent and a second solution; mixing the combined red cells and first solutions and the combined pathogen treatment agent and the second solution to provide a combination; and incubating the combination.

Claim 115. The method of Claim 114 further comprising combining the second part solution with a quenching agent.

Claim 116. The method of Claim 114 wherein the pathogen treatment agent is unreconstituted and the second solution reconstitutes the agent.

Claim 117. The method of Claim 116 in which the pathogen treatment agent is dry.

Claim 118. The method of Claim 115 in which the combination is agitated during incubation.

Claim 119. The method of Claim 115 comprising contacting the combination with a sorbtion media.

Claim 120. The method of Claim 115 in which the mixing is carried out statically.

Claim 121. The method of Claim 114 in which the first solution comprises a solution having a pH comparable to the pH of said red blood cells.

Claim 122. The method of Claim 114 in which the second solution has a pH that is acidic.

Claim 123. The method of Claim 114 wherein the incubation continues for a plurality of hours.

Claim 124. The method of Claim 119 in which the combination is contacted with the sorption media for a plurality of hours.

Claim 125. The method of Claim 114 in which the pathogen treatment agent is relatively stable or inactive at a pH that is relatively acidic compared to the normal pH of red cells and is activated upon combination with red cells.

Claim 126. The method of Claim 125 in which the quenching agent is combined with pathogen treatment agent prior to the contact with red cells.

Claim 127. The method of Claim 114 in which the first part of the storage solution comprises sodium citrate, one or more buffers, adenine and mannitol, said solution being substantially free of chloride and sugars.

Claim 128. The method of Claim 127 in which said buffers include sodium phosphate monobasic and sodium phosphate dibasic.

Claim 129. The method of Claim 114 in which the second part comprises a sugar selected from the group consisting of fructose and dextrose.

Claim 130. The method of Claim 114 in which the first part of the storage solution comprises approximately 1 mMol/1 to about 2.2 mMol/1 adenine, approximately 20 mMol/1 to about 110 mMol/1 mannitol, approximately 2.2 mMol/1 to about 90 mMol/1 sodium citrate, approximately 1 mMol/1 to about 10 mMol/1 sodium phosphate/monobasic and approximately 5 mMol/1 to about 25 mMol/1 sodium phosphate dibasic.

Claim 131. The method of Claim 114 in which the pathogen treatment agent comprises a pathogen inactivation agent.

Claim 132. The method of Claim 131 in which the pathogen inactivation agent prevents and/or reduces replication of bacterial and viral pathogens .

Claim 133. A method for ex-vivo inactivation of collected red cells suspected of containing pathogens comprising: combining the red cells and a first solution; combining a pathogen inactivation agent and a second solution; combining a quenching agent and the second solution; combining the red cells, first solution, pathogen inactivation agent, second solution, and quenching agent; and incubating the combined red cells, first part, pathogen inactivation agent, second solution and quenching agent.

Claim 134. The method of Claim 133 in which the pathogen inactivation agent is unreconstituted and the method further comprises reconstituting the pathogen inactivation agent.

Claim 135. The method of Claim 133 in which the quenching agent is unreconstituted and the method further comprises reconstituting the quenching agent.

Claim 136. The method of Claim 134 including removing unused pathogen inactivation agent, a degradation product of the agent and/or quenching agent after incubation.

Claim 137. A system for the ex-vivo treatment of collected red cells suspected of containing pathogens comprising: means for combining the red cells and a first solution; means for combining a pathogen treatment agent and a second solution; means for mixing the combined red cells and first solution and the combined pathogen treatment agent and the second solution to provide a combination; and means for incubating the combination.

Claim 138. The system of Claim 137 further comprising means for combining the second solution with a quenching agent .

Claim 139. The system of Claim 137 wherein the pathogen treatment agent is unreconstituted and the second solution reconstitutes the agent.

Claim 140. The system of Claim 137 in which the pathogen treatment agent is dry.

Claim 141. The system of Claim 137 including means agitate the combination during incubation.

Claim 142. The system of Claim 137 further comprising a sorption media and means for contacting the combination with the sorption media.

Claim 143. The system of Claim 137 in which the means for mixing comprises a static mixer.

Claim 144. The system of Claim 137 in which the first solution has a pH comparable to the pH of said red blood cells .

Claim 145. The system of Claim 137 in which the second solution has a pH that is acidic. <

Claim 146. The system of Claim 137 in which the pathogen treatment agent is relatively stable or inactive at a pH that is relatively acidic compared to the normal pH of red cells and is activated upon combination with red cells.

Claim 147. The system of Claim 138 in which the means for combining the quenching agent with the pathogen treatment agent combines them prior to the contact with red cells.

Claim 148. The system of Claim 137 in which the first solution comprises sodium citrate, one or more buffers, adenine and mannitol, said solution being substantially free of chloride and sugars .

Claim 149. The system of Claim 138 in which said buffers include sodium phosphate monobasic and sodium phosphate dibasic .

Claim 150. The system of Claim 137 in which the second solution comprises a sugar selected from the group consisting of fructose and dextrose.

Claim 151. The system of Claim 137 in which the first solution comprises approximately 1 mMol/1 to about 2.2 mMol/1 adenine, approximately 20 mMol/1 to about 110 mMol/1 mannitol, approximately 2.2 mMol/1 to about 90 mMol/1 sodium citrate, approximately 1 mMol/1 to about 10 mMol/1 sodium phosphate/monobasic and approximately 5 mMol/1 to about 25 mMol/1 sodium phosphate dibasic.

Claim 152. The system of Claim 137 in which the pathogen treatment agent comprises a pathogen inactivation agent.

Claim 153. The system of Claim 152 in which the pathogen inactivation agent prevents replication of bacterial and viral pathogens .