WO2002058849A1 - Multi-well filtration apparatus - Google Patents

Multi-well filtration apparatus Download PDF

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Publication number
WO2002058849A1
WO2002058849A1 PCT/US2002/000594 US0200594W WO02058849A1 WO 2002058849 A1 WO2002058849 A1 WO 2002058849A1 US 0200594 W US0200594 W US 0200594W WO 02058849 A1 WO02058849 A1 WO 02058849A1
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WO
WIPO (PCT)
Prior art keywords
vacuum
plate
well
collection
collection plate
Prior art date
Application number
PCT/US2002/000594
Other languages
French (fr)
Inventor
Roger Q. Roberts
Michael Wuelk
Original Assignee
Varian, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Varian, Inc. filed Critical Varian, Inc.
Priority to AU2002243493A priority Critical patent/AU2002243493B2/en
Priority to EP02708982A priority patent/EP1360009B1/en
Priority to DE60216418T priority patent/DE60216418T2/en
Priority to JP2002559175A priority patent/JP4117191B2/en
Publication of WO2002058849A1 publication Critical patent/WO2002058849A1/en

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B01L3/50255—Multi-well filtration
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/18—Apparatus therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/20—Accessories; Auxiliary operations
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/2575—Volumetric liquid transfer

Definitions

  • the present invention generally relates to filtration apparatus and more specifically relates to a multi-well filtration or solid phase extraction apparatus .
  • Multi-well filtration/solid phase extraction assemblies are well known in the art and are used for the assay of biological liquid specimens.
  • Conventional filtration assemblies typically comprise a filtration plate, or separation plate, having multiple wells for receiving a liquid specimen, and a collection tray having a plurality of wells for collecting filtrate.
  • the separation plate and the collection tray are disposed in a stacked relationship such that individual collection wells are associated each with a single filtration well.
  • a conventional multi-well filtration plate has 96 wells for performing multiple assays simultaneously.
  • Each well typically contains a separations media, for example a filter member, for separating a biological component from the liquid that is introduced into the separation plate, and allowing a liquid portion of the biological fluid to filter into the collection tray.
  • a multi-well filtration/solid phase extraction assembly requires vacuum, positive pressure, centrifugation or other mechanical means to force the liquid through the separations media.
  • a vacuum box useful for vacuum processing multiple biological samples in a multi-well filtration plate is described in U.S. Patent No. 5,205,989 and U.S. Patent No. 5,283,039, both to Aysta.
  • the Aysta vacuum box and other conventional vacuum assemblies that are designed to process multiple samples simultaneously are costly, thereby discouraging many users from adopting the 96- well format for sample processing.
  • the present invention provides a low cost interface between a multi-well separation plate and the collection plate for enabling vacuum processing of multiple samples without need for an expensive vacuum assembly.
  • the present invention provides effective means for simultaneously vacuum processing multiple fluid samples without the need for a more expensive, conventional vacuum box.
  • the present apparatus generally comprises a unique vacuum collar that is adapted to be connected to, and in an interfacing relationship with, a 96-well separation plate and a 96- well collection plate.
  • the separation plate generally includes a plurality of filtration wells each including an inlet portion, and a relatively narrow outlet portion forming a nozzle-like tip.
  • the filtration wells extend through a depth or thickness of the separation plate.
  • a separation media is provided within each of the filtration wells for filtering a biological fluid sample to be assayed.
  • the collection plate includes a plurality of collection wells for receiving filtrate or eluate that is discharged from an associated one of the filtration wells .
  • the vacuum collar when interconnected between the separation plate and the collection plate, defines a substantially air-tight chamber.
  • Means are provided for connecting the chamber to a vacuum source, such as an inexpensive vacuum pump assembly.
  • a vacuum source such as an inexpensive vacuum pump assembly.
  • a hose barb connector extending through the collar and adapted to be connected to a vacuum hose is provided.
  • means are provided for enabling manual control of pressure differential in the chamber in order to suit a particular application.
  • the vacuum collar includes means, for example a keyed structure, for ensuring alignment between separation wells and collection wells.
  • the vacuum collar is adapted to place each separation well discharge tip a specified distance into each collection well to ensure that eluates or filtrates will not contaminate adjoining collection wells.
  • the vacuum collar may include other features which ensure proper connection of the separation plate and the collection plate each time the apparatus is used, thereby eliminating possibility of error in eluate or filtrate transfer.
  • a multi-well filtration apparatus in which the collection plate is adapted to be directly connected to a vacuum assembly in a stacked configuration. More specifically, the collection plate includes means, including a vent opening sized and structured to receive an air-intake member of the vacuum assembly when the collection means is disposed on the vacuum assembly in a stacked relationship.
  • Figure 1 shows a perspective view of a multi-well filtration apparatus in accordance with the present invention
  • Figure 2 shows an exploded view of the multi-well filtration apparatus shown in Figure 1;
  • Figure 3 shows a top view of the multi-well filtration apparatus shown in Figure 1;
  • Figure 4 shows a cross-sectional view of the apparatus taken across line 4-4 of Figure 3 ;
  • Figure 5 shows a partially cut away side view of another multi-well filtering apparatus in accordance with the present invention.
  • Figure 6 shows a collection plate of the multi-well filtering apparatus shown in Figure 5.
  • Figure 7 shows a cross sectional view of the apparatus taken across line 7-7 of Figure 5.
  • the apparatus 10 generally includes a separation plate 12 adapted to receive a fluid, for example a blood specimen or other biological fluid, a vacuum collar 14, and collection means 16 for capturing and containing components of the biological fluid passed through the separation plate 12.
  • Means, for example a hose barb connector 18, for connecting the vacuum collar 14 to a vacuum source (not shown) is also included.
  • the separation plate 12, shown most clearly in Figure 2 may comprise a conventional, multi-well separation plate.
  • the separation plate 12 includes means, including a plurality of wells 22, for example 96 wells, for receiving multiple test samples of a biological fluid to be processed by filtration or solid phase extraction techniques.
  • the separation plate wells 22 will hereinafter sometimes be referred to as "filtration wells" . It will be appreciated that the number of wells 22 found in the separation plate 12 in the embodiment 10 shown is simply a matter of convenience for the analyst or investigator.
  • the separation plate 12 may contain as few as one well, or as many wells as are functionally permissible to give the actual dimensions of the plate 12. Typical test samples of a biological fluid are often less than about 500 microliters in volume, and the wells 22 are sized appropriately to contain these small volumes.
  • the separation plate 12 may be formed of any resilient and non-reactive material that is commonly available.
  • a separation media 24 is disposed generally at a bottom of each of the filtration wells 22 and functions to separate and retain components of the biological fluid as the fluid passes through the well 22.
  • the separation media 24 comprises any filter, membrane, matrix or the like, in a single layer or multiple layers thereof, that is suitable for the application on hand. Attention is directed specifically to Figure 4.
  • the separation plate 12 may be a substantially unitary, molded structure with each of the filtration wells 22 comprising a generally cylindrical aperture 34 extending through a full thickness, or depth, of the separation plate 12. More specifically, each filtration well 22 includes a relatively wide receiving inlet portion 38 and relatively smaller outlet portion 39 forming a nozzle-like tip.
  • the filter 24 is disposed at the bottom of the inlet portion 38 as shown, and may be held in place by a retaining ring 42 or other conventional means.
  • the inlet portion 38 of adjoining wells 22 may be defined by a common walls 44.
  • the separation plate 12 includes a substantially rectangular upper portion 46 having a face 47 in which the filtration wells 22 form a matrix arrangement as shown.
  • the separation plate 12 further includes a lower portion 48 from which the nozzle-like tips 39 of the filtration wells 22 depend.
  • the collection means 16 in the shown embodiment 10 comprises a multi- well collection plate.
  • the collection plate 16 includes means, for example a plurality of wells 52, for receiving a liquid component, i.e. filtrate or eluate, of the biological fluid sample which is discharged from outlet tips 39 of the filtration wells 22.
  • Each of the collection wells 52 has an inlet 53 defined in a face 54
  • the collection plate 16 may include any number of such wells 52, with a 96-well collection plate being a typical example. Adjoining collection wells 52 may share a common wall 58.
  • the vacuum collar 14 is adapted to interface and interconnect the separation plate 12 and the collection plate 16. More specifically, as shown in Figure 2, the vacuum collar 14 includes inner wall 62 defining a generally central space 64, and an outer wall 66 . Space 64 is sized to receive the lower portion 48 of the separation plate 12 and the face 54 of the collection plate 16.
  • each one of the collection wells 52 is aligned with an associated one of the filtration wells 22.
  • the vacuum collar 14 is adapted to position the outlet tips 39 of the filtration wells 22 a specified distance into the collection wells 52, thereby insuring that filtrates or eluates will not contaminate adjoining wells.
  • a chamber 68 to which a vacuum can be applied is defined between the separation plate 12, collection plate 16, and inner wall 62; see Figure 4.
  • the vacuum collar 14 may include a valve 70 in communication with the chamber 68.
  • the valve 70 may be used to facilitate venting of the system as needed, through port 72, and may be opened and closed by means of handle 76.
  • the valve 70 may include a second port 78, comprising for example a female Luer fitting, for enabling serial connection of multiple multi-well filtration apparatuses 10 in accordance with the invention.
  • the port 78 can be utilized as a means for interconnecting the vacuum collar 14 with another vacuum collar in accordance with the invention may be connected and operated by means of a single vacuum source.
  • Cap 79 is provided for sealing the second port 78 when not in use. It is to be appreciated that although only one specific arrangement is shown, there are many different possible valve arrangements for providing vacuum to the system in accordance with the invention, using inexpensive valves and/or hose barb connectors.
  • the apparatus 10 is assembled as shown in Figures 1 and 4. Specimen samples are deposited in a conventional manner into the filtration wells 22.
  • a vacuum source (not shown), connected to a hose fitting 78 of the vacuum connector 18 is activated.
  • a valve member 70 (see Figures 1 and 3) is placed in an open position. Air in the chamber 68 is drawn through air intake 72 (see Figure 4) and a partial vacuum is produced within the chamber 68. This partial vacuum will assist in drawing biological fluid within in each filtration well 22 through the filter 24 or other separation media. The filtered fluid, or "filtrate” will drip into the corresponding collection well 52.
  • the collar 14 in accordance with the invention may include a valve specifically for controlling a pressure differential in the chamber such that depending upon the application, the differential in the chamber 68 can be adjusted between a low pressure differential and a relatively higher pressure differential .
  • the vacuum collar 14 may further include means for ensuring that the separation plate 22 and the collection plate 16 are always aligned in the same direction. Attention is directed again to Figures 1 and 3 , wherein it is shown that the separation plate 12 includes a beveled area 95 on the rectangular portion 46 that will only fit in the vacuum collar 14 when positioned properly within keyed area 96. Similarly, as shown in Figure 2, the collection plate 16 includes a beveled area 97 on its rectangular portion 16 that will only fit within the vacuum collar when positioned properly to the vacuum collar's keyed area 98. These keyed areas 96 and 98 of the vacuum collar 14 will only accept the separation plate 12 and the collection plate 16 in one orientation ensuring proper alignment of the separation plate 12 and the collection plate 16.
  • this embodiment 100 generally comprises a multi-well separation plate 12, a vacuum collar 114, a collection plate 116, and a vacuum assembly 118 in a stacked relationship.
  • the vacuum assembly 118 may include comprise a housing 119, an internal vacuum mechanism, more specifically a vacuum pump 120, disposed in the hosing 119 and an air intake member 122 extending upward from the housing 119.
  • the housing 119 may include a recessed upper surface 124 configured to contain the collection plate 116 as shown.
  • the vacuum pump 120 may be any suitable vacuum pump, for example such as a vacuum pump available from KNF Neuberger, Inc.
  • the air intake member 122 may extend from the recessed surface of the housing as shown and may be elongated or slot-like in shape .
  • the collection plate 116 includes means, for example vent opening 126, for receiving the air intake member 122 when the collection plate 116 is stacked on the recessed surface 124 of the vacuum housing 119.
  • the vent opening 126 preferably conforms to the dimensions of the air intake member 122 and extends across a substantial portion of a width of the collection plate 116 as shown. This design provides a substantially uniform vacuum distribution across the chamber 168.
  • the vacuum collar 114 may be identical to the collar 14 described hereinabove. However, it will be appreciated that in this embodiment 100, the vacuum collar 114 does not require a separate vacuum connector 18 or other vacuum connections or manifolds. Air is drawn from a vacuum chamber 168, (defined between the vacuum collar 114, collection plate 116 and separation plate 12) , directly through the air intake member 122 received in the vent opening 126 of the collection plate 116.
  • this embodiment 100 provides vacuum processing of multiple biological fluid samples, through the stacking of the separation plate 12, collection plate 114 and vacuum assembly 118 in a towerlike relationship. It can be appreciated that this embodiment 100 does not require a substantial area of table or counter space to operate.
  • this embodiment 100 does not require a substantial area of table or counter space to operate.

Abstract

A multi-well filtration (10) or solid phase extraction apparatus is provided which generally includes a vacuum collar (14) adapted to be interconnected between a conventional 96-well sample application plate, or separation plate (12), and a 96-well collection plate (16). The vacuum collar provides an interface chamber between these plates through which a vacuum can be applied to assist in the filtration which a vacuum can be applied to assist in the filtration process. The collection plate may include a vent opening providing a direct communication between the interface chamber and a vacuum assembly air intake, when the collection plate is disposed on top of the vacuum assembly in a stacked configuration, thereby eliminating any need for separate vacuum manifolds or connectors.

Description

MULTI-WELL FILTRATION APPARATUS
The present invention generally relates to filtration apparatus and more specifically relates to a multi-well filtration or solid phase extraction apparatus .
Multi-well filtration/solid phase extraction assemblies are well known in the art and are used for the assay of biological liquid specimens. Conventional filtration assemblies typically comprise a filtration plate, or separation plate, having multiple wells for receiving a liquid specimen, and a collection tray having a plurality of wells for collecting filtrate. The separation plate and the collection tray are disposed in a stacked relationship such that individual collection wells are associated each with a single filtration well. A conventional multi-well filtration plate has 96 wells for performing multiple assays simultaneously. Each well typically contains a separations media, for example a filter member, for separating a biological component from the liquid that is introduced into the separation plate, and allowing a liquid portion of the biological fluid to filter into the collection tray.
For many applications, a multi-well filtration/solid phase extraction assembly requires vacuum, positive pressure, centrifugation or other mechanical means to force the liquid through the separations media. A vacuum box useful for vacuum processing multiple biological samples in a multi-well filtration plate is described in U.S. Patent No. 5,205,989 and U.S. Patent No. 5,283,039, both to Aysta. Unfortunately, the Aysta vacuum box and other conventional vacuum assemblies that are designed to process multiple samples simultaneously, are costly, thereby discouraging many users from adopting the 96- well format for sample processing.
The present invention provides a low cost interface between a multi-well separation plate and the collection plate for enabling vacuum processing of multiple samples without need for an expensive vacuum assembly.
Summary of the Invention
Accordingly, a multi-well filtration and solid phase extraction apparatus is provided by the present invention which is useful for facilitating vacuum processing of liquid biological samples using conventional multi-well separation plates and filtrate or waste collection plates. Advantageously, the present invention provides effective means for simultaneously vacuum processing multiple fluid samples without the need for a more expensive, conventional vacuum box. In one particular embodiment, the present apparatus generally comprises a unique vacuum collar that is adapted to be connected to, and in an interfacing relationship with, a 96-well separation plate and a 96- well collection plate.
The separation plate generally includes a plurality of filtration wells each including an inlet portion, and a relatively narrow outlet portion forming a nozzle-like tip. The filtration wells extend through a depth or thickness of the separation plate. A separation media is provided within each of the filtration wells for filtering a biological fluid sample to be assayed. Similarly, the collection plate includes a plurality of collection wells for receiving filtrate or eluate that is discharged from an associated one of the filtration wells .
Importantly, the vacuum collar, when interconnected between the separation plate and the collection plate, defines a substantially air-tight chamber. Means are provided for connecting the chamber to a vacuum source, such as an inexpensive vacuum pump assembly. For example, a hose barb connector extending through the collar and adapted to be connected to a vacuum hose is provided. In addition, means are provided for enabling manual control of pressure differential in the chamber in order to suit a particular application.
Preferably, the vacuum collar includes means, for example a keyed structure, for ensuring alignment between separation wells and collection wells. In another aspect of the invention, the vacuum collar is adapted to place each separation well discharge tip a specified distance into each collection well to ensure that eluates or filtrates will not contaminate adjoining collection wells. In addition, the vacuum collar may include other features which ensure proper connection of the separation plate and the collection plate each time the apparatus is used, thereby eliminating possibility of error in eluate or filtrate transfer.
In one particular embodiment of the invention, a multi-well filtration apparatus is provided in which the collection plate is adapted to be directly connected to a vacuum assembly in a stacked configuration. More specifically, the collection plate includes means, including a vent opening sized and structured to receive an air-intake member of the vacuum assembly when the collection means is disposed on the vacuum assembly in a stacked relationship.
Brief Description of the Drawings
The present invention will be more clearly understood and the objects and advantages better appreciated by referring to the following detailed description considered in conjunction with the accompanying drawings of which:
Figure 1 shows a perspective view of a multi-well filtration apparatus in accordance with the present invention; Figure 2 shows an exploded view of the multi-well filtration apparatus shown in Figure 1;
Figure 3 shows a top view of the multi-well filtration apparatus shown in Figure 1;
Figure 4 shows a cross-sectional view of the apparatus taken across line 4-4 of Figure 3 ;
Figure 5 shows a partially cut away side view of another multi-well filtering apparatus in accordance with the present invention;
Figure 6 shows a collection plate of the multi-well filtering apparatus shown in Figure 5; and
Figure 7 shows a cross sectional view of the apparatus taken across line 7-7 of Figure 5.
Detailed Description
Turning now to Figures 1-3, a multi-well filtration apparatus 10 in accordance with the present invention is shown. The apparatus 10 generally includes a separation plate 12 adapted to receive a fluid, for example a blood specimen or other biological fluid, a vacuum collar 14, and collection means 16 for capturing and containing components of the biological fluid passed through the separation plate 12. Means, for example a hose barb connector 18, for connecting the vacuum collar 14 to a vacuum source (not shown) is also included.
The separation plate 12, shown most clearly in Figure 2, may comprise a conventional, multi-well separation plate. The separation plate 12 includes means, including a plurality of wells 22, for example 96 wells, for receiving multiple test samples of a biological fluid to be processed by filtration or solid phase extraction techniques. The separation plate wells 22 will hereinafter sometimes be referred to as "filtration wells" . It will be appreciated that the number of wells 22 found in the separation plate 12 in the embodiment 10 shown is simply a matter of convenience for the analyst or investigator. The separation plate 12 may contain as few as one well, or as many wells as are functionally permissible to give the actual dimensions of the plate 12. Typical test samples of a biological fluid are often less than about 500 microliters in volume, and the wells 22 are sized appropriately to contain these small volumes. The separation plate 12 may be formed of any resilient and non-reactive material that is commonly available.
As shown in a cut-away portion of Figure 2, a separation media 24 is disposed generally at a bottom of each of the filtration wells 22 and functions to separate and retain components of the biological fluid as the fluid passes through the well 22. The separation media 24 comprises any filter, membrane, matrix or the like, in a single layer or multiple layers thereof, that is suitable for the application on hand. Attention is directed specifically to Figure 4. The separation plate 12 may be a substantially unitary, molded structure with each of the filtration wells 22 comprising a generally cylindrical aperture 34 extending through a full thickness, or depth, of the separation plate 12. More specifically, each filtration well 22 includes a relatively wide receiving inlet portion 38 and relatively smaller outlet portion 39 forming a nozzle-like tip. The filter 24 is disposed at the bottom of the inlet portion 38 as shown, and may be held in place by a retaining ring 42 or other conventional means. The inlet portion 38 of adjoining wells 22 may be defined by a common walls 44. Referring now to Figures 1 - 3, the separation plate 12 includes a substantially rectangular upper portion 46 having a face 47 in which the filtration wells 22 form a matrix arrangement as shown. Turning specifically to Figure 2, the separation plate 12 further includes a lower portion 48 from which the nozzle-like tips 39 of the filtration wells 22 depend.
Referring now to Figures 2 and 4, the collection means 16 in the shown embodiment 10 comprises a multi- well collection plate. The collection plate 16 includes means, for example a plurality of wells 52, for receiving a liquid component, i.e. filtrate or eluate, of the biological fluid sample which is discharged from outlet tips 39 of the filtration wells 22. Each of the collection wells 52 has an inlet 53 defined in a face 54
(see Figure 2) of the collection plate 16, and a closed bottom 56 (see Figure 4) for containing the liquid component. The collection plate 16 may include any number of such wells 52, with a 96-well collection plate being a typical example. Adjoining collection wells 52 may share a common wall 58.
Importantly, in accordance with the present invention, the vacuum collar 14 is adapted to interface and interconnect the separation plate 12 and the collection plate 16. More specifically, as shown in Figure 2, the vacuum collar 14 includes inner wall 62 defining a generally central space 64, and an outer wall 66 . Space 64 is sized to receive the lower portion 48 of the separation plate 12 and the face 54 of the collection plate 16.
When the vacuum collar 14 is interconnected between the separation plate 12 and the collection plate 16, each one of the collection wells 52 is aligned with an associated one of the filtration wells 22. In the shown embodiment, the vacuum collar 14 is adapted to position the outlet tips 39 of the filtration wells 22 a specified distance into the collection wells 52, thereby insuring that filtrates or eluates will not contaminate adjoining wells.
A chamber 68 to which a vacuum can be applied is defined between the separation plate 12, collection plate 16, and inner wall 62; see Figure 4. In addition to the hose barb fitting 18, the vacuum collar 14 may include a valve 70 in communication with the chamber 68.
The valve 70 may be used to facilitate venting of the system as needed, through port 72, and may be opened and closed by means of handle 76. In addition, turning back to Figure 1, in another aspect of the invention, the valve 70 may include a second port 78, comprising for example a female Luer fitting, for enabling serial connection of multiple multi-well filtration apparatuses 10 in accordance with the invention. Advantageously, the port 78 can be utilized as a means for interconnecting the vacuum collar 14 with another vacuum collar in accordance with the invention may be connected and operated by means of a single vacuum source. Cap 79 is provided for sealing the second port 78 when not in use. It is to be appreciated that although only one specific arrangement is shown, there are many different possible valve arrangements for providing vacuum to the system in accordance with the invention, using inexpensive valves and/or hose barb connectors.
In operation, the apparatus 10 is assembled as shown in Figures 1 and 4. Specimen samples are deposited in a conventional manner into the filtration wells 22. A vacuum source (not shown), connected to a hose fitting 78 of the vacuum connector 18 is activated.
A valve member 70 (see Figures 1 and 3) is placed in an open position. Air in the chamber 68 is drawn through air intake 72 (see Figure 4) and a partial vacuum is produced within the chamber 68. This partial vacuum will assist in drawing biological fluid within in each filtration well 22 through the filter 24 or other separation media. The filtered fluid, or "filtrate" will drip into the corresponding collection well 52.
Although not shown, the collar 14 in accordance with the invention may include a valve specifically for controlling a pressure differential in the chamber such that depending upon the application, the differential in the chamber 68 can be adjusted between a low pressure differential and a relatively higher pressure differential .
It is also noted that the vacuum collar 14 may further include means for ensuring that the separation plate 22 and the collection plate 16 are always aligned in the same direction. Attention is directed again to Figures 1 and 3 , wherein it is shown that the separation plate 12 includes a beveled area 95 on the rectangular portion 46 that will only fit in the vacuum collar 14 when positioned properly within keyed area 96. Similarly, as shown in Figure 2, the collection plate 16 includes a beveled area 97 on its rectangular portion 16 that will only fit within the vacuum collar when positioned properly to the vacuum collar's keyed area 98. These keyed areas 96 and 98 of the vacuum collar 14 will only accept the separation plate 12 and the collection plate 16 in one orientation ensuring proper alignment of the separation plate 12 and the collection plate 16.
Turning now to Figures 5-7 another multi-well filtration apparatus 100 in accordance with the present the invention is shown, with features similar or identical to those described hereinabove having like reference numbers for the sake of convenience. More particularly, this embodiment 100 generally comprises a multi-well separation plate 12, a vacuum collar 114, a collection plate 116, and a vacuum assembly 118 in a stacked relationship.
The vacuum assembly 118 may include comprise a housing 119, an internal vacuum mechanism, more specifically a vacuum pump 120, disposed in the hosing 119 and an air intake member 122 extending upward from the housing 119. The housing 119 may include a recessed upper surface 124 configured to contain the collection plate 116 as shown. The vacuum pump 120 may be any suitable vacuum pump, for example such as a vacuum pump available from KNF Neuberger, Inc. The air intake member 122 may extend from the recessed surface of the housing as shown and may be elongated or slot-like in shape .
The collection plate 116 (see Figure 6) includes means, for example vent opening 126, for receiving the air intake member 122 when the collection plate 116 is stacked on the recessed surface 124 of the vacuum housing 119. The vent opening 126 preferably conforms to the dimensions of the air intake member 122 and extends across a substantial portion of a width of the collection plate 116 as shown. This design provides a substantially uniform vacuum distribution across the chamber 168.
The vacuum collar 114 may be identical to the collar 14 described hereinabove. However, it will be appreciated that in this embodiment 100, the vacuum collar 114 does not require a separate vacuum connector 18 or other vacuum connections or manifolds. Air is drawn from a vacuum chamber 168, (defined between the vacuum collar 114, collection plate 116 and separation plate 12) , directly through the air intake member 122 received in the vent opening 126 of the collection plate 116.
Advantageously, this embodiment 100 provides vacuum processing of multiple biological fluid samples, through the stacking of the separation plate 12, collection plate 114 and vacuum assembly 118 in a towerlike relationship. It can be appreciated that this embodiment 100 does not require a substantial area of table or counter space to operate. Although there has been hereinabove described a multi-well filtration apparatus, in accordance with the present invention, for purposes of illustrating the manner in which the invention may be used to advantage, it will be appreciated that the invention is not limited thereto. Accordingly, any and all modifications, variations, or equivalent arrangements which may occur to those skilled in the art should be considered to be within the scope of the present invention as defined in the appended claims .

Claims

What is claimed is:
1. A multi-well filtration/solid phase extraction apparatus for use with a multi-well separation plate and a multi-well collection plate, the apparatus comprising: a vacuum collar adapted to be connected to both a multi-well separation plate and to a multi-well collection plate, the vacuum collar including a chamber, wherein the chamber provides a substantially air tight interface between the multi-well separation plate and the multi-well collection plate when the vacuum collar is connected to both the multi-well separation plate and the multi-well collection plate; and a connector for connecting the vacuum collar chamber to a vacuum source in order to deliver vacuum within the collection plate.
2. The apparatus according to claim 1 wherein the vacuum collar is adapted to be connected to a conventional ninety six well separation plate.
3. The apparatus according to claim 2 wherein the vacuum collar is further adapted to be connected to a conventional ninety-six well collection plate.
4. The apparatus according to claim 1 further comprising keyed structure for ensuring appropriate alignment between the separation plate and the collection plate to reduce possibility of error in eluate or filtrate transfer.
5. The apparatus according to claim 1 further comprising a port, disposed on the vacuum collar, for enabling interconnection of the vacuum collar with another vacuum collar to enable plurality of multi-well filtration/solid phase extraction apparatus to be operated from a single vacuum source.
6. A multi-well filtration/solid phase extraction apparatus comprising: a separation plate including multiple separation wells for receiving and filtering a biological fluid sample; a collection plate including multiple collection wells for receiving filtrate from the multiple separation wells; a vacuum collar adapted to be interconnected between the separation plate and the collection plate so as to define a substantially air-tight chamber between the separation plate and the collection plate; and a connector, disposed on the vacuum collar, for connection the vacuum collar chamber to vacuum source in order to deliver vacuum with in the collection plate .
7. The apparatus according to claim 6 wherein the multiple separation wells include about ninety-six wells.
8. The apparatus according to claim 6 wherein the multiple collection wells includes about ninety-six multiple collection wells defined in the collection plate.
9. The apparatus according to claim 6 further comprising keyed structure for ensuring appropriate alignment between the separation plate and the collection plate to reduce possibility of error in eluate or filtrate transfer.
10. The apparatus according to claim 6 further comprising a port, disposed on the vacuum collar, for enabling interconnection of the vacuum collar with another vacuum collar to enable a plurality of multi- well filtration/solid phase extraction apparatus to be operated from a single vacuum source.
11. A multi-well filtration/solid phase extraction apparatus comprising: a separation plate including multiple separation wells for receiving and separating a biological fluid specimen; collection plate, including multiple collection wells for receiving a liquid component of the biological fluid specimen, the collection plate including a vent opening; a collar for connecting the collection plate with the separation plate in an interfacing relationship; and a vacuum assembly including a housing adapted to receive the collection plate in a stacked configuration therewith, and air intake member configured to be received by the vent opening of the collection plate, and vacuum mechanism for drawing air through the vent opening and air intake member when the collection plate is disposed on the vacuum assembly in the stacked configuration to enable vacuum processing of the biological fluid specimen received in the separation plate .
12. The apparatus according to claim 11 wherein the vacuum mechanism comprises a vacuum pump disposed in the vacuum assembly housing.
13. The apparatus according to claim 11 wherein the separation plate comprises a ninety-six well separation plate.
14. The apparatus according to claim 13 wherein the collection plate comprises a ninety-six well collection plate.
PCT/US2002/000594 2001-01-23 2002-01-09 Multi-well filtration apparatus WO2002058849A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU2002243493A AU2002243493B2 (en) 2001-01-23 2002-01-09 Multi-well filtration apparatus
EP02708982A EP1360009B1 (en) 2001-01-23 2002-01-09 Multi-well filtration apparatus
DE60216418T DE60216418T2 (en) 2001-01-23 2002-01-09 MORE TANK FILTRATION DEVICE
JP2002559175A JP4117191B2 (en) 2001-01-23 2002-01-09 Multiwell filtration device

Applications Claiming Priority (2)

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US09/767,500 2001-01-23
US09/767,500 US6491873B2 (en) 2001-01-23 2001-01-23 Multi-well filtration apparatus

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WO2002058849A1 true WO2002058849A1 (en) 2002-08-01

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US (1) US6491873B2 (en)
EP (1) EP1360009B1 (en)
JP (1) JP4117191B2 (en)
AU (1) AU2002243493B2 (en)
DE (1) DE60216418T2 (en)
ES (1) ES2274960T3 (en)
WO (1) WO2002058849A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004024950A1 (en) * 2002-09-10 2004-03-25 Noxxon Pharma Ag Method for the selection of nucleic acid ligands

Families Citing this family (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6270730B1 (en) * 1998-06-16 2001-08-07 Northwest Engineering Inc. Multi-well rotary synthesizer
GB9906477D0 (en) * 1999-03-19 1999-05-12 Pyrosequencing Ab Liquid dispensing apparatus
AU2002217141A1 (en) * 2000-12-20 2002-07-01 Dsm Ip Assets B.V. Preparation of fluid samples by applying pressure
US6899848B1 (en) * 2001-02-27 2005-05-31 Hamilton Company Automated sample treatment system: apparatus and method
US20020182118A1 (en) * 2001-05-31 2002-12-05 Perry Brian A. Vacuum manifold for both multi-well plate and individual columns
US6682703B2 (en) * 2001-09-05 2004-01-27 Irm, Llc Parallel reaction devices
KR100411946B1 (en) * 2001-12-18 2003-12-18 한국해양연구원 Device for simultaneously collecting filtered water and filter paper
US20030143124A1 (en) * 2002-01-31 2003-07-31 Roberts Roger Q. Unidirectional flow control sealing matt
US20030148537A1 (en) * 2002-02-06 2003-08-07 Franck Bellon Mask for depositing and distributing reagents on an analytical support
US7048893B2 (en) * 2002-02-06 2006-05-23 Sebia Mask for depositing and distributing reagents on an analytical support
US20030202909A1 (en) * 2002-04-29 2003-10-30 Genetix Limited Vacuum manifold and uses thereof
DE50300434D1 (en) * 2002-05-31 2005-05-19 Tecan Trading Ag Maennedorf Apparatus, system and method for aspirating liquids from solid phase extraction plates
US7122155B2 (en) * 2002-07-16 2006-10-17 Mcgill University Electron microscopy cell fraction sample preparation robot
US20040129676A1 (en) * 2003-01-07 2004-07-08 Tan Roy H. Apparatus for transfer of an array of liquids and methods for manufacturing same
EP1491258B1 (en) * 2003-06-24 2008-12-03 Millipore Corporation Multifunctional vacuum manifold
US7629140B2 (en) * 2004-10-07 2009-12-08 Istituto Di Richerche Di Biologia Molecolare P. Angeletti S.P.A. Assay for cytochrome P450 isoform 2C9
EP1799842A1 (en) * 2004-10-07 2007-06-27 Istituto Di Ricerche Di Biologia Molecolare P. Angeletti S.P.A. Assay for cytochrome p450 isoforms 3a4 and 3a5
US7563410B2 (en) * 2004-10-19 2009-07-21 Agilent Technologies, Inc. Solid phase extraction apparatus and method
JP5123847B2 (en) * 2005-05-25 2013-01-23 バイオ−ラド ラボラトリーズ インコーポレイテッド Fluid engineering equipment
CN101312785B (en) * 2005-09-02 2011-03-30 罗什生命科学公司 Systems, apparatus and methods for vacuum filtration
JP2007212285A (en) * 2006-02-09 2007-08-23 Enplas Corp Fluid handling device
US20070202607A1 (en) * 2006-02-24 2007-08-30 Revesz Robert N Filtration apparatus and associated method for microwave-assisted chemistry
EP1854540A1 (en) * 2006-05-12 2007-11-14 F. Hoffmann-la Roche AG Multi-well filtration device
US8158009B2 (en) * 2007-05-23 2012-04-17 Roush Life Sciences, Llc Methods and apparatus for foam control in a vacuum filtration system
US8231012B2 (en) * 2007-07-26 2012-07-31 Roush Life Sciences, Llc Filtrate storage system
EP2183040B1 (en) * 2007-07-26 2012-03-07 Roush Life Sciences, LLC Methods and apparatus for supporting a vacuum filtration device
JP2011209084A (en) * 2010-03-30 2011-10-20 Sumitomo Bakelite Co Ltd Microplate
US8663580B2 (en) 2010-11-01 2014-03-04 Agilent Technologies, Inc. Dried biological fluid spot punch device and related methods
US9005543B2 (en) 2010-11-01 2015-04-14 Agilent Technologies, Inc. Apparatus for punching and solid phase extraction of dried biological fluid spot and related methods
ES2358699B1 (en) 2011-03-09 2012-03-14 Zf Biotox, S.L. MICROPLACE FOR BIOLOGICAL TESTS.
US20130264286A1 (en) * 2012-04-06 2013-10-10 Cells Scientific Corporation Biological sample filtering system and method for filtering biological samples
JP2014069149A (en) * 2012-09-28 2014-04-21 Sumitomo Bakelite Co Ltd Spacer and processing unit
EP2972319B1 (en) 2013-03-11 2019-02-20 Becton, Dickinson and Company Bacterial detection cartridge
WO2015011522A1 (en) 2013-07-23 2015-01-29 Tubitak Ume Multi-task sample preparation system with reconfigurable modules for on-line dilution, enzymatic digestion and fractionation
WO2015078884A1 (en) * 2013-11-26 2015-06-04 Alleati Ag Method and microfluidic assembly for antibiotic susceptibility testing
US9568404B2 (en) 2014-05-16 2017-02-14 Junyu Mai Method and apparatus for biomolecule analysis
JP6956203B2 (en) 2017-05-10 2021-11-02 イー・エム・デイー・ミリポア・コーポレイシヨン Multi-well plate with variable compression seal
EP3593903B1 (en) * 2018-07-10 2020-09-02 Sartorius Stedim Biotech GmbH Liquid filtering system with vacuum valve
US11724261B1 (en) * 2019-05-13 2023-08-15 Integrated Dna Technologies, Inc. Synthesizer system with inflatable seal and valve arrangement

Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4090850A (en) * 1976-11-01 1978-05-23 E. R. Squibb & Sons, Inc. Apparatus for use in radioimmunoassays
US4301010A (en) * 1980-03-10 1981-11-17 Spectrum Medical Industries, Inc. Vacuum filter
US4797259A (en) * 1986-12-15 1989-01-10 Pall Corporation Well-type diagnostic plate device
US4895706A (en) * 1986-10-28 1990-01-23 Costar Corporation Multi-well filter strip and composite assemblies
US4902481A (en) * 1987-12-11 1990-02-20 Millipore Corporation Multi-well filtration test apparatus
US4927604A (en) * 1988-12-05 1990-05-22 Costar Corporation Multiwell filter plate vacuum manifold assembly
US4948564A (en) * 1986-10-28 1990-08-14 Costar Corporation Multi-well filter strip and composite assemblies
US5219528A (en) * 1989-07-28 1993-06-15 Pierce Chemical Company Apparatus for rapid immunoassays
US5227137A (en) * 1991-04-04 1993-07-13 Nicholson Precision Instruments Inc. Vacuum clamped multi-sample filtration apparatus
US5603900A (en) * 1995-05-19 1997-02-18 Millipore Investment Holdings Limited Vacuum filter device
US5792430A (en) * 1996-08-12 1998-08-11 Monsanto Company Solid phase organic synthesis device with pressure-regulated manifold
US6133045A (en) * 1998-02-27 2000-10-17 Hamilton Company Automated sample treatment system: apparatus and method
US6159368A (en) * 1998-10-29 2000-12-12 The Perkin-Elmer Corporation Multi-well microfiltration apparatus
US6309605B1 (en) * 1999-05-05 2001-10-30 Millipore Corporation Well(s) containing filtration devices
US6331431B1 (en) * 1995-11-28 2001-12-18 Ixsys, Inc. Vacuum device and method for isolating periplasmic fraction from cells

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4734192A (en) 1982-07-01 1988-03-29 Millipore Corporation Multiwell membrane filtration apparatus
US5108704A (en) * 1988-09-16 1992-04-28 W. R. Grace & Co.-Conn. Microfiltration apparatus with radially spaced nozzles
US5205989A (en) 1991-09-18 1993-04-27 Minnesota Mining And Manufacturing Company Multi-well filtration apparatus
EP0991928A2 (en) * 1997-06-27 2000-04-12 Immunetics Rapid flow-through binding assay apparatus and method
US6455007B1 (en) * 2000-06-13 2002-09-24 Symyx Technologies, Inc. Apparatus and method for testing compositions in contact with a porous medium

Patent Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4090850A (en) * 1976-11-01 1978-05-23 E. R. Squibb & Sons, Inc. Apparatus for use in radioimmunoassays
US4301010A (en) * 1980-03-10 1981-11-17 Spectrum Medical Industries, Inc. Vacuum filter
US4895706A (en) * 1986-10-28 1990-01-23 Costar Corporation Multi-well filter strip and composite assemblies
US4948564A (en) * 1986-10-28 1990-08-14 Costar Corporation Multi-well filter strip and composite assemblies
US4797259A (en) * 1986-12-15 1989-01-10 Pall Corporation Well-type diagnostic plate device
US4902481A (en) * 1987-12-11 1990-02-20 Millipore Corporation Multi-well filtration test apparatus
US4927604A (en) * 1988-12-05 1990-05-22 Costar Corporation Multiwell filter plate vacuum manifold assembly
US5219528A (en) * 1989-07-28 1993-06-15 Pierce Chemical Company Apparatus for rapid immunoassays
US5227137A (en) * 1991-04-04 1993-07-13 Nicholson Precision Instruments Inc. Vacuum clamped multi-sample filtration apparatus
US5603900A (en) * 1995-05-19 1997-02-18 Millipore Investment Holdings Limited Vacuum filter device
US5792425A (en) * 1995-05-19 1998-08-11 Millipore Coporation Vacuum filter device
US6331431B1 (en) * 1995-11-28 2001-12-18 Ixsys, Inc. Vacuum device and method for isolating periplasmic fraction from cells
US5792430A (en) * 1996-08-12 1998-08-11 Monsanto Company Solid phase organic synthesis device with pressure-regulated manifold
US6133045A (en) * 1998-02-27 2000-10-17 Hamilton Company Automated sample treatment system: apparatus and method
US6159368A (en) * 1998-10-29 2000-12-12 The Perkin-Elmer Corporation Multi-well microfiltration apparatus
US6338802B1 (en) * 1998-10-29 2002-01-15 Pe Corporation (Ny) Multi-well microfiltration apparatus
US6309605B1 (en) * 1999-05-05 2001-10-30 Millipore Corporation Well(s) containing filtration devices

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP1360009A4 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004024950A1 (en) * 2002-09-10 2004-03-25 Noxxon Pharma Ag Method for the selection of nucleic acid ligands

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AU2002243493B2 (en) 2006-03-09
JP4117191B2 (en) 2008-07-16
ES2274960T3 (en) 2007-06-01
DE60216418D1 (en) 2007-01-11
EP1360009A1 (en) 2003-11-12
EP1360009A4 (en) 2005-11-09
DE60216418T2 (en) 2007-09-27
US6491873B2 (en) 2002-12-10
JP2004526950A (en) 2004-09-02
US20020098125A1 (en) 2002-07-25

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