CA2108292C - Process for preparing hepatitis a (hav) antigens and vaccines - Google Patents
Process for preparing hepatitis a (hav) antigens and vaccines Download PDFInfo
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- CA2108292C CA2108292C CA002108292A CA2108292A CA2108292C CA 2108292 C CA2108292 C CA 2108292C CA 002108292 A CA002108292 A CA 002108292A CA 2108292 A CA2108292 A CA 2108292A CA 2108292 C CA2108292 C CA 2108292C
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- detergent
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- gel filtration
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/32011—Picornaviridae
- C12N2770/32411—Hepatovirus, i.e. hepatitis A virus
- C12N2770/32422—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/32011—Picornaviridae
- C12N2770/32411—Hepatovirus, i.e. hepatitis A virus
- C12N2770/32434—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/948—Microorganisms using viruses or cell lines
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Communicable Diseases (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Process for preparing hepatitis A (HAV) antigens and vaccines.
The HAV virus is multiplied on competent cells, the infected cells are lysed, the supernatant is recovered and the purification is carried out by a chromatographic procedure on an anion-exchange support and a gel filtration procedure, the purification procedures being carried out in the presence of a detergent, and the chromatographic procedure being carried out under conditions which retain the virions or viral capsids, which are then eluted.
The HAV virus is multiplied on competent cells, the infected cells are lysed, the supernatant is recovered and the purification is carried out by a chromatographic procedure on an anion-exchange support and a gel filtration procedure, the purification procedures being carried out in the presence of a detergent, and the chromatographic procedure being carried out under conditions which retain the virions or viral capsids, which are then eluted.
Description
- ~1~D~~9~
The present invention relates to a new process for preparing hepatitis A virus (HAV) antigens and vaccines.
The preparation of vaccines which are effective against hepatitis A is now known. On the other hand, advances remain to be made in the industrial field of culture of the HAV virus and purification of the capsid antigens of this picornavirus.
The initial processes for the partial purification of I3AV virions having proved too expensive or unsatisfactory for the production of vaccines, application EP-A-0,302,692 recommended subjecting the cells having served for the multiplication of the HAV
virus, lysed beforehand, to organic solvent extraction procedures combined with polyethylene glycol (PEG) precipitations. The extract may then be optionally subjected to a chromatography on an anion-exchange support allowing the viral capsids to pass through, followed by a gel filtration. This process is presented as having the advantage of avoiding any use of detergents.
Subsequently, application EP-A-0,468,702 combined with this same process a second chromatographic step on an anion-exchange support under conditions which make it possible to retain the viral capsids which are then eluted, the eluate being subjected to the gel filtration step.
These processes have the disadvantage of being complicated and requiring the use of organic solvents.
The present invention proposes to overcome the disadvantages of the prior art and to provide a new process for preparing hepatitis A antigens and vaccines. -The subject of the invention is a process for preparing hepatitis A antigens and vaccines in which the HAV virus is multiplied on competent cells, the infected cells are lysed preferably - by sonication, the supernatant is recovered and the purification is carried out by a chromatographic procedure on an anion-exchange support and a gel filtration procedure, characterized in that the purification procedures' are carried out in the presence of a detergent, preferably Tween 80, and in that the chromatographic procedure is carried out under conditions which retain the virions or viral capsids, which are they eluted.
In the present invention, the presence of the detergent is especially designed to avoid the phenomena of adsorption of the capsids or virions. The Tween 80 (sorbitaa mono-9-octadecaaoate) is remarkably effective, preferably at concentrations ~f 0.001 to 5%. especially of 0.1%. Consequently, the invention also relates to the use of detergents. which, °.for this lunation, are equivalea~ to Tai~e:i 80.
The use of the detergent according to the invention makes it possible to avoid the extraction steps of the prior art and to carry out. after simply filtering the lyeis supernatant, a single ion-exchange chromatographic step on a support which retains the viral capsids whereas the protein and nucleic contamia,ants are a0 removed in the effluent. The antigenic capsids are then easily recovered by elution.
Preferably. the chromatographic support consists of D8AE-Spherodex* (Sepracor* IBF DSA$-Spherodex LS Ref.
61002) but supports such as Pharmva~aia DEAF-Sepharose*can also be used. Preferably, the chromatographic column is ec;uilibrated with buffer. containing the detergent, especially Tween 80; the eluent may also contain the detergent.
The eluate is. preferably -concentrated after the chroanatography, especially by ultrafiltration.
The eluate, preferably concentrated, is then subjected to the gel filtration, preferably on Sepharose 6B CL. in the presence of the detergent.
The viral peak obtained can then be again coaceatsated and then inactivated by~a customary process.
Other advantages and characteristics of the invention will emerge on reading the following description, made by way of non-restrictive example.
*Trade-mark Example ' 3 1) Culture of the cells.
The cells used .a=e humsa deploid cells of the MRCS line. The working stock consists of the 16th passage.
The vial containing the working stock is thawed and transferred into a culture flask F.75 cm= into which the cell culture medium is iatraduced fa the presence of foetal calf serum (FCS)-and the culture dish is placed is as oven at 39°C. Whey the cells are confluent is the culture dishes,- a trypsini~atioa procedure is carried out.
The culture medium is removed from the culture container: the cell layer is rinsed with a PHS solution and the cells are detached by a dilute trypsia solution, after which cell culture medium is agaia~iatroduced, the cells are dispersed in the medium and the cellular suspension obtained is distributed into the new culture containers. The culture is they carried out from the 18th to the 38th passage wsiag iizcreasingly larger containers.
for example by going-from a 75 cm~ flask during the 18th passac~ up to .the CF 40 caultitray container using, for intermediate passages, the customary culture containers cosrespoading to the increasing volumes to-be treated.
After 24 h of cultur~a at 37°C the culture medium of the CF 40 containers of the 38th passage is removed by aspiration and replaced by sa iaoculum consisting of cell culture medium containing foetal calf sermmi cad a quantity of hepatitis A virus so as to have a multiplicity of is~featioa of between 0 : Ol and - 1. 8ach CF 40 receives 8 litres of this medium.
The CF 40 containers are then replaced in the oven (temperature of 35°C) cad the viral propagation is carried out is 21 days with medium changes after 8 days of culture with medium containing 5~ calf serum, they of ter 16 days with FCS-free medium. The virus is harvested after 21 days of culture.
The present invention relates to a new process for preparing hepatitis A virus (HAV) antigens and vaccines.
The preparation of vaccines which are effective against hepatitis A is now known. On the other hand, advances remain to be made in the industrial field of culture of the HAV virus and purification of the capsid antigens of this picornavirus.
The initial processes for the partial purification of I3AV virions having proved too expensive or unsatisfactory for the production of vaccines, application EP-A-0,302,692 recommended subjecting the cells having served for the multiplication of the HAV
virus, lysed beforehand, to organic solvent extraction procedures combined with polyethylene glycol (PEG) precipitations. The extract may then be optionally subjected to a chromatography on an anion-exchange support allowing the viral capsids to pass through, followed by a gel filtration. This process is presented as having the advantage of avoiding any use of detergents.
Subsequently, application EP-A-0,468,702 combined with this same process a second chromatographic step on an anion-exchange support under conditions which make it possible to retain the viral capsids which are then eluted, the eluate being subjected to the gel filtration step.
These processes have the disadvantage of being complicated and requiring the use of organic solvents.
The present invention proposes to overcome the disadvantages of the prior art and to provide a new process for preparing hepatitis A antigens and vaccines. -The subject of the invention is a process for preparing hepatitis A antigens and vaccines in which the HAV virus is multiplied on competent cells, the infected cells are lysed preferably - by sonication, the supernatant is recovered and the purification is carried out by a chromatographic procedure on an anion-exchange support and a gel filtration procedure, characterized in that the purification procedures' are carried out in the presence of a detergent, preferably Tween 80, and in that the chromatographic procedure is carried out under conditions which retain the virions or viral capsids, which are they eluted.
In the present invention, the presence of the detergent is especially designed to avoid the phenomena of adsorption of the capsids or virions. The Tween 80 (sorbitaa mono-9-octadecaaoate) is remarkably effective, preferably at concentrations ~f 0.001 to 5%. especially of 0.1%. Consequently, the invention also relates to the use of detergents. which, °.for this lunation, are equivalea~ to Tai~e:i 80.
The use of the detergent according to the invention makes it possible to avoid the extraction steps of the prior art and to carry out. after simply filtering the lyeis supernatant, a single ion-exchange chromatographic step on a support which retains the viral capsids whereas the protein and nucleic contamia,ants are a0 removed in the effluent. The antigenic capsids are then easily recovered by elution.
Preferably. the chromatographic support consists of D8AE-Spherodex* (Sepracor* IBF DSA$-Spherodex LS Ref.
61002) but supports such as Pharmva~aia DEAF-Sepharose*can also be used. Preferably, the chromatographic column is ec;uilibrated with buffer. containing the detergent, especially Tween 80; the eluent may also contain the detergent.
The eluate is. preferably -concentrated after the chroanatography, especially by ultrafiltration.
The eluate, preferably concentrated, is then subjected to the gel filtration, preferably on Sepharose 6B CL. in the presence of the detergent.
The viral peak obtained can then be again coaceatsated and then inactivated by~a customary process.
Other advantages and characteristics of the invention will emerge on reading the following description, made by way of non-restrictive example.
*Trade-mark Example ' 3 1) Culture of the cells.
The cells used .a=e humsa deploid cells of the MRCS line. The working stock consists of the 16th passage.
The vial containing the working stock is thawed and transferred into a culture flask F.75 cm= into which the cell culture medium is iatraduced fa the presence of foetal calf serum (FCS)-and the culture dish is placed is as oven at 39°C. Whey the cells are confluent is the culture dishes,- a trypsini~atioa procedure is carried out.
The culture medium is removed from the culture container: the cell layer is rinsed with a PHS solution and the cells are detached by a dilute trypsia solution, after which cell culture medium is agaia~iatroduced, the cells are dispersed in the medium and the cellular suspension obtained is distributed into the new culture containers. The culture is they carried out from the 18th to the 38th passage wsiag iizcreasingly larger containers.
for example by going-from a 75 cm~ flask during the 18th passac~ up to .the CF 40 caultitray container using, for intermediate passages, the customary culture containers cosrespoading to the increasing volumes to-be treated.
After 24 h of cultur~a at 37°C the culture medium of the CF 40 containers of the 38th passage is removed by aspiration and replaced by sa iaoculum consisting of cell culture medium containing foetal calf sermmi cad a quantity of hepatitis A virus so as to have a multiplicity of is~featioa of between 0 : Ol and - 1. 8ach CF 40 receives 8 litres of this medium.
The CF 40 containers are then replaced in the oven (temperature of 35°C) cad the viral propagation is carried out is 21 days with medium changes after 8 days of culture with medium containing 5~ calf serum, they of ter 16 days with FCS-free medium. The virus is harvested after 21 days of culture.
3) Crude harvest On the day of the harvest, the CF 40 containers are carefully examined and possible suspect CF 40 ~~~&29~
.w. - 4 -containers are eliminated.
The supernatant culture liquid is removed.
The cellular layer is rinsed twice with two litres of PBS solution.
The cellular layer is then rinsed with 2000 ml of trypsin-EDTA solution.
The CF 40 containers are replaced in the oven at 35°C for 5 minutes.
1000 ml of 20 mM phosphate buffered solution, pH 7.50, are poured into each CF container, the CF
containers are shaken so as to detach all the cells from the culture surface and the cellular suspension is then recovered,. The CF containers are then rinsed twice with 1 litre of phosphate buffer.
The cellular suspension recovered (volume of about 40 litres for 12 CF 40 containers) is homogenized and treated continuously with ultrasound at a frequency of 20 KHz in an incubation tank with a flow rate of between 0.5 and 10 ml/second.
After treatment, the cellular suspension is centrifuged at low speed for 30 minutes at 2000 rpm.
The supernatant recovered constitutes the crude harvest.
.w. - 4 -containers are eliminated.
The supernatant culture liquid is removed.
The cellular layer is rinsed twice with two litres of PBS solution.
The cellular layer is then rinsed with 2000 ml of trypsin-EDTA solution.
The CF 40 containers are replaced in the oven at 35°C for 5 minutes.
1000 ml of 20 mM phosphate buffered solution, pH 7.50, are poured into each CF container, the CF
containers are shaken so as to detach all the cells from the culture surface and the cellular suspension is then recovered,. The CF containers are then rinsed twice with 1 litre of phosphate buffer.
The cellular suspension recovered (volume of about 40 litres for 12 CF 40 containers) is homogenized and treated continuously with ultrasound at a frequency of 20 KHz in an incubation tank with a flow rate of between 0.5 and 10 ml/second.
After treatment, the cellular suspension is centrifuged at low speed for 30 minutes at 2000 rpm.
The supernatant recovered constitutes the crude harvest.
4) Addition of the detergent After removing control samples, a Tween 80 solution is added to the crude harvest tank so as to adjust the final Tween 80 content to 0.1~.
The solution is left -stirring overnight at + 4°C.
The harvest is then filtered through 0.45 and 0.2 ~. Durapore-type filters.
This solution therefore constitutes the filtered harvest.
The solution is left -stirring overnight at + 4°C.
The harvest is then filtered through 0.45 and 0.2 ~. Durapore-type filters.
This solution therefore constitutes the filtered harvest.
5) Chromatography The DEAF-Spherodex gel is loaded into a column with O.1N HC1 and prepared by passing 20 mM phosphate buffer, pH 7.5, then a solution of 2 ~...formalin 2 g/1 NaCl, and then 1 M NaCl.
- - - In order to equilibrate the column and then carry out the chromatography, 20 mM P04 buffer, pH 7.5, 0.1% Tween 80 is passed until perfect equilibration of pH and osmolality are obtained; the sample, whose osmolality should not exceed 280 mOsm, is passed; 20 mM P04 buffer, 0.5 M NaCl pH 7.5, 0.1% Tween 80 is passed. This procedure allows elution of the virus.
The flow rate is of the order of 45 cm/h.
- - - In order to equilibrate the column and then carry out the chromatography, 20 mM P04 buffer, pH 7.5, 0.1% Tween 80 is passed until perfect equilibration of pH and osmolality are obtained; the sample, whose osmolality should not exceed 280 mOsm, is passed; 20 mM P04 buffer, 0.5 M NaCl pH 7.5, 0.1% Tween 80 is passed. This procedure allows elution of the virus.
The flow rate is of the order of 45 cm/h.
6) Concentration The material used is a Fellicon*-type ultrafiltration cell equipped with Sartorius ULTRASART* II
cassettes made from cellulose ester (molecular weight cut-off 10,000 daltons).
The entire installation, cassette, module and piping is sterilized by means of a 5% hydrogen peroxide solution.
The entire installation is rinsed with 20 litres of 20 mM phosphate buffer pH = 7.5, 0.1% Tween 80.
The eluate from the DEAE-Spherodex column, fraction in 20 mM phosphate, 0.5 M NaCl, 0.1% Tween 80, containing the virus is concentrated.
The final volume after rinsing the membranes is adjusted to 1 litre.
cassettes made from cellulose ester (molecular weight cut-off 10,000 daltons).
The entire installation, cassette, module and piping is sterilized by means of a 5% hydrogen peroxide solution.
The entire installation is rinsed with 20 litres of 20 mM phosphate buffer pH = 7.5, 0.1% Tween 80.
The eluate from the DEAE-Spherodex column, fraction in 20 mM phosphate, 0.5 M NaCl, 0.1% Tween 80, containing the virus is concentrated.
The final volume after rinsing the membranes is adjusted to 1 litre.
7) Gel filtration A Pharmacies K215/100 column, filled with 26.5 litres of Sepharose 6B CL gel, is used. Gel height: 73 cm.
The entire installation column, piping, detector, is sterilized by passing a 2% formalin solution, 2% NaCl, *Trade-mark _ 5a _ flow rate 1.7 1/h.
The column is washed and equilibrated by means of 20 mM phosphate solution pH = 7.5, 0.1~ Tween 80 for 48 hours, flow rate: 2.7 1/h.
The concentrated virus is introduced into the column by gravity; the purification is carried out by means of 20 mM phosphate buffer pH = 7.5, 0.1~ Tween 80, flow rate: 2.7 1/h.
The entire installation column, piping, detector, is sterilized by passing a 2% formalin solution, 2% NaCl, *Trade-mark _ 5a _ flow rate 1.7 1/h.
The column is washed and equilibrated by means of 20 mM phosphate solution pH = 7.5, 0.1~ Tween 80 for 48 hours, flow rate: 2.7 1/h.
The concentrated virus is introduced into the column by gravity; the purification is carried out by means of 20 mM phosphate buffer pH = 7.5, 0.1~ Tween 80, flow rate: 2.7 1/h.
8) Final concentration Under conditions similar to those used during the preceding concentration, the virus peak derived from the gel filtration colu~maa is concentrated. The final volume after rinsing is adjusted to a.5 litres. The concentrated virus is filtered through a 0.2 ~C Durapore*membraae.
This product correapoads to then purified harvest.
This product correapoads to then purified harvest.
9) Iaactivation_ The inactivation is carried out using formalin at 1/4000 for 14 d at 3?°C.
10) Preparation of the vaccine The vaccine is preaduced from the inactivated purified harvest, adsorbed cm~to aluminium hydro~i.de, the packaging being carried out ace:ordiag to conventional methods.
*Trade-mark
*Trade-mark
Claims (10)
1. Process for preparing hepatitis A antigens and vaccines in which the HAV virus is multiplied on competent cells, the infected cells are lysed, the supernatant is recovered and the purification is carried out successively by a chromatographic procedure on an anion-exchange support and a gel filtration procedure, wherein said purification is carried out in the presence of a detergent, and said chromatographic procedure is carried out under conditions which retain the virions or viral capsids, which are then eluted.
2. Process according to Claim 1, characterized in that Tween* 80 is used as detergent.
3. Process according to one of Claims 1 and 2, characterized in that the detergent is used at concentrations of 0.001 to 5%.
4. Process according to one of Claims 1 and 2, characterized in that the detergent is used at a concentration of 0.1%.
5. Process according to any one of Claims 1 to 4, characterized in that the cells are lysed by sonication.
6. Process according to any one of Claims 1 to 5, characterized in that the chromatographic procedure is carried out on an anion-exchange support through which a buffer containing detergent is passed, then an elution is carried out using an eluent also containing said detergent.
7. Process according to any one of Claims 1 to 6, characterized in that the gel filtration procedure is carried out on a column equilibrated by means of a buffer containing said detergent.
8. Process according to any one of Claims 1 to 7, characterized in that a concentration procedure is carried out between the chromatographic procedure and the gel filtration procedure.
9. Process according to any one of Claims 1 to 8, characterized in that a final concentration procedure is carried out after the gel filtration procedure, after which the preparation of antigens is inactivated.
10. Process according to any one of Claims 1 to 9, characterized in that the lysis is performed by sonication at a frequency of the order of 20,000 Hz.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR9212285A FR2696748B1 (en) | 1992-10-14 | 1992-10-14 | Process for the preparation of hepatitis A antigens and vaccines (HAV). |
| FR9212285 | 1992-10-14 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2108292A1 CA2108292A1 (en) | 1994-04-15 |
| CA2108292C true CA2108292C (en) | 2005-04-12 |
Family
ID=9434534
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002108292A Expired - Lifetime CA2108292C (en) | 1992-10-14 | 1993-10-13 | Process for preparing hepatitis a (hav) antigens and vaccines |
Country Status (10)
| Country | Link |
|---|---|
| US (1) | US5731187A (en) |
| EP (1) | EP0593339B1 (en) |
| JP (1) | JP3724821B2 (en) |
| AT (1) | ATE179428T1 (en) |
| CA (1) | CA2108292C (en) |
| DE (1) | DE69324645T2 (en) |
| DK (1) | DK0593339T3 (en) |
| ES (1) | ES2132204T3 (en) |
| FR (1) | FR2696748B1 (en) |
| GR (1) | GR3030636T3 (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU6908598A (en) * | 1996-10-29 | 1998-05-22 | Smithkline Beecham Biologicals (Sa) | Purification of respiratory syncytial virus antigens |
| US6544769B1 (en) | 1996-12-13 | 2003-04-08 | Schering Corporation | Compostions comprising viruses and methods for concentrating virus preparations |
| US6261823B1 (en) | 1996-12-13 | 2001-07-17 | Schering Corporation | Methods for purifying viruses |
| JP2001506126A (en) * | 1996-12-13 | 2001-05-15 | シェーリング コーポレイション | Virus purification method |
| CA2482512C (en) * | 2002-04-30 | 2011-09-20 | Oncolytics Biotech Inc. | Improved viral purification methods |
| CN101018804A (en) * | 2004-07-27 | 2007-08-15 | 基诺美生物工程（制药）公司 | Method of purifying virus envelope |
| US7901921B2 (en) * | 2004-10-22 | 2011-03-08 | Oncolytics Biotech Inc. | Viral purification methods |
| IL246769B (en) * | 2015-07-23 | 2020-09-30 | Grifols Sa | Methods for purification of a virus produced in vitro and clearance assay for the virus |
| EP3695849A1 (en) * | 2019-02-13 | 2020-08-19 | Mediagnost Gesellschaft für Forschung und Herstellung von Diagnostika GmbH | Method for extracting hepatitis a virus (hav) antigen from cell culture |
| KR102277089B1 (en) * | 2019-12-19 | 2021-07-14 | 에스케이바이오사이언스(주) | Method for Preparing Hepatitis A Virus and Hepatitis A Virus Prepared by the Method |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NZ225583A (en) * | 1987-08-06 | 1991-07-26 | Merck & Co Inc | Process for purifying hepatitis a virions (hav) |
| JPH0761955B2 (en) * | 1988-04-28 | 1995-07-05 | 国立予防衛生研究所長 | Lyophilized hepatitis A vaccine |
| CA2047041A1 (en) * | 1990-07-18 | 1992-01-19 | John A. Lewis | Process for purification of hepatitis-a virus capsids |
| IT1248075B (en) * | 1991-06-18 | 1995-01-05 | Sclavo Spa | HEPATITIS A (HAV) VIRUS PURIFICATION PROCESS, PURIFIED VIRUS AND VACCINAL COMPOSITIONS THAT CONTAIN IT. |
-
1992
- 1992-10-14 FR FR9212285A patent/FR2696748B1/en not_active Expired - Fee Related
-
1993
- 1993-10-07 DK DK93402469T patent/DK0593339T3/en active
- 1993-10-07 DE DE69324645T patent/DE69324645T2/en not_active Expired - Lifetime
- 1993-10-07 AT AT93402469T patent/ATE179428T1/en active
- 1993-10-07 ES ES93402469T patent/ES2132204T3/en not_active Expired - Lifetime
- 1993-10-07 EP EP93402469A patent/EP0593339B1/en not_active Expired - Lifetime
- 1993-10-13 CA CA002108292A patent/CA2108292C/en not_active Expired - Lifetime
- 1993-10-13 US US08/136,580 patent/US5731187A/en not_active Expired - Lifetime
- 1993-10-14 JP JP28050093A patent/JP3724821B2/en not_active Expired - Fee Related
-
1999
- 1999-06-30 GR GR990401722T patent/GR3030636T3/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| FR2696748B1 (en) | 1994-12-30 |
| US5731187A (en) | 1998-03-24 |
| GR3030636T3 (en) | 1999-10-29 |
| CA2108292A1 (en) | 1994-04-15 |
| ATE179428T1 (en) | 1999-05-15 |
| JP3724821B2 (en) | 2005-12-07 |
| DE69324645D1 (en) | 1999-06-02 |
| DE69324645T2 (en) | 1999-12-30 |
| JPH06279317A (en) | 1994-10-04 |
| FR2696748A1 (en) | 1994-04-15 |
| ES2132204T3 (en) | 1999-08-16 |
| EP0593339A1 (en) | 1994-04-20 |
| DK0593339T3 (en) | 1999-11-08 |
| EP0593339B1 (en) | 1999-04-28 |
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