CA2175670C - Process for producing a virus-inactivated factor viii-containing fraction by chromatographic methods - Google Patents
Process for producing a virus-inactivated factor viii-containing fraction by chromatographic methods Download PDFInfo
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- CA2175670C CA2175670C CA002175670A CA2175670A CA2175670C CA 2175670 C CA2175670 C CA 2175670C CA 002175670 A CA002175670 A CA 002175670A CA 2175670 A CA2175670 A CA 2175670A CA 2175670 C CA2175670 C CA 2175670C
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- process according
- factor viii
- ionic strength
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- sodium chloride
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/36—Blood coagulation or fibrinolysis factors
- A61K38/37—Factors VIII
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/745—Blood coagulation or fibrinolysis factors
- C07K14/755—Factors VIII, e.g. factor VIII C (AHF), factor VIII Ag (VWF)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
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- Pharmacology & Pharmacy (AREA)
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- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
A process for producing a virus-inactivated, factor VIII-containing fraction by chromatographic methods in which, starting with cryoprecipitate or blood plasma, possibly followed by a treatment with aluminium hydroxide, at least one separation operating is performed by membrane chromatography after the dissolution of the cryoprecipitate. The fractions are subjected to virus inactivation and preferably an additional pasteurisation step.
Description
A Process for the Preparation of a Virus Inactivated Fraction Containing Factor VIII by means of Chromatographic Methods This inventions is related to a process for the preparation of a virus inactivated fraction containing factor VIII by means of chromatographic methods.
Factor VIII is a vital material that plays an important role in blood clotting. Blood clotting disorders can thus be treated by administration of factor VIII. Therefore, a large need exists for administrable factor VIII preparations.
Numerous attempts have been made to isolate factor VIII in a highly enriched form from natural sources. Thus, chromatographic methods for the purification of factor VIII
from cryoprecipitates are already known. The latter is a fraction which is available by treatment of plasma in the cold. EP 367 840 B1 pertains to a chromatographic method for the isolation of factor VIII from blood plasma without a preliminary cryoprecipitation. The fraction containing factor VIII is separated by chromatographic separation on a hydrophilic chromatographic material modified by ion-exchanging groups . EP 0 238 701 pertains to a method for the preparation of an ultra-pure, infectious antihemophilic factor, where the pretreated fractions are a cryoprecipitate liberated from fibrinogen, globulin, albumins, and other interfering ingredients by means of ethanol precipitation.
EP-A-0 343 275 describes chromatographic separation with ion exchangers after virus inactivation of the cryoprecipitate fraction. EP 0 173 242 A describes a method for obtaining factor VIII preparations by chromatography on anion-exchanging materials which are solely based on carbohydrates, the carbohydrate matrix being modified by DEAF groups. In particular, DEAF sepharose and DEAF cellulose are described as being useful. In GB-A-1,178,958, a purification of factor VIII
using ECTEOLA cellulose columns is described. The modified cellulose contains basic substituents introduced by reaction of epichlorohydrin and triethanolamine. The above mentioned prior art makes use either of chromatographic separation in the form of batch processes or of column chromatography.
Although fairly good results are obtained with these methods, it still remains desirable for economic as well as ethical reasons to increase the yield of biologically valuable factor VIII.
Hence, the technical problem underlying this invention consists in providing a method which, proceeding from the prior art, allows for an improved preparation of factor VIII
with respect to yield and biological activity.
Surprisingly, the problem has been solved by a process proceeding from a cryoprecipitate after dissolution thereof or blood plasma, optionally followed by an aluminum hydroxide treatment, and employing a separation step using membrane chromatography on membranes or compact disks of the porous hydrophilic polymers poly(glycidyl methacrylates) or hydrophilized polystyrene.
The process of the invention can be performed with commercial cryoprecipitate or blood plasma. Preferably, the thawed cryoprecipitate is treated with aluminium hydroxide for further pre-purification of the sample in order to preconcentrate factor VIII.
Preferably, a virus inactivation is carried out prior to the actual chromatographic purification on materials arranged within or on membranes. This virus inactivation is performed according to the method described in EP 131 740 A1 by treatment with biocompatible organic solvents (detergents), Triton~ X-100/TNBP, preferably Tween~/TNBP (tri-n-butyl phosphate). Good results will also be obtained with sodium cholate/TNPB. Preferably, detergents are employed in amounts of up to 15o by weight. Wherein a virus inactivation is performed by treatment of said fraction with a di- or trialkyl phosphate and a non-ionic surfactant.
The chromatographic separation step for the purification of factor VIII in the sample may be carried out either on base materials modified by ion-exchanging groups, particularly anion exchangers, or on materials modified by immunoaffinity ligands. It is critical that the mentioned materials are arranged in membranes. Particularly useful are membranes as well as compact disks made of porous poly(glycidyl methacrylates) and/or hydrophilized polystyrene.
A membrane suitable for separation consists of compact disks made of polymer carriers. The base materials of the membranes or disks are provided with corresponding anion-exchanging groups or immunoaffinity ligands. Particularly anion-exchanging groups such as quaternary amines or diethylaminoethyl groups are considered as ion-exchanging groups. Suitable ca n on exchangers are in principle weakly and strongly acidic ration exchangers, such as materials which are modified by sulfonic acid or phosphoric acid groups.
The ion-exchanging groups can be bound to the base material fibers with or without a so-called spacer.
Materials provided with spacers are also referred to as tentacle materials. In DE 42 04 694 corresponding spacers and ligands are mentioned. For example, a glucosamine residue can serve as a spacer as well. Anion-exchanging groups such as DEAF or quaternary amines may also be bound at the membranes made of porous poly(glycidyl methacrylate) or the other materials mentioned. The binding of the anion-exchanging groups takes place either directly to the material forming the membrane or else through a spacer as well, e.g. a glucosamine residue.
In another embodiment of the process of the invention affinity membrane chromatography with immobilized substances having a high affinity for factor VIII is used.
The substances having affinity for factor VIII are immobilized at the carrier by means of chemically reactive groups. Preferably, the reactive group will attack at the end of a spacer rather than directly at the carrier material. Immobilization of the substances for factor VIII
takes place through binding at reactive groups such as tosyl, tresyl, hydrazid and others. Corresponding procedures are known from T. M. Phillips "Affinity chromatography" in "Chromatography" (E. Heftmann, ed.), 5th ed. Elsevier, Amsterdam 1992.
In another preferred embodiment materials are employed for the separation of factor VIII that can ensure a hydrophobic interaction. Hydrophobic materials which are employed are acyclic and/or cyclic alkyl chains, for instance C1 to C18 alkyl chains, and aromatic substances.
Suitable materials providing hydrophobic interaction preferably include those having graded hydrophobicity.
Hydrophobicity can be graded by introduction of polar groups, such as protic-polar or aprotic-polar groups, for example hydroxy, amino, cyano groups. Preferably, it is adapted corresponding to the respective separation conditions.
Virus inactivation may also take place by heat treatment. Preferably, the eluated sample containing factor VIII is subjected to a pasteurization step following a first membrane chromatographic step. A corresponding procedure is proposed in DE-A-4 318 435. Therein, fraction enriched with factor VIII are contacted with di- or trialkyl phosphates and optionally wetting agents in the presence of stabilizing agents and simultaneously or subsequently treated for a period of 5 hours to 30 hours at elevated temperatures ranging from 55°C to 67°C. It may be of advantage to combine the two methods of virus inactivation, the treatment with detergents and with heat.
In order to remove the chemicals employed in the pasteurization step, a second membrane chromatography may follow. Preferably, the separation of the added stabilizing agents takes place by means of a membrane modified by DEAF or quaternary ammonium compounds that are arranged on the surface of the chromatographic carrier material through a spacer. In addition, it is possible to arrange the corresponding ligands on the surface of the carrier material without a spacer. The stabilizing agents are not retarded by this anion exchange material under the conditions chosen, whereas factor VIII is adsorbed on the chromatographic material.
Thereafter, factor VIII is eluated with an aqueous solvent system exhibiting gradually increasing salt concentrations.
Factor VIII is a vital material that plays an important role in blood clotting. Blood clotting disorders can thus be treated by administration of factor VIII. Therefore, a large need exists for administrable factor VIII preparations.
Numerous attempts have been made to isolate factor VIII in a highly enriched form from natural sources. Thus, chromatographic methods for the purification of factor VIII
from cryoprecipitates are already known. The latter is a fraction which is available by treatment of plasma in the cold. EP 367 840 B1 pertains to a chromatographic method for the isolation of factor VIII from blood plasma without a preliminary cryoprecipitation. The fraction containing factor VIII is separated by chromatographic separation on a hydrophilic chromatographic material modified by ion-exchanging groups . EP 0 238 701 pertains to a method for the preparation of an ultra-pure, infectious antihemophilic factor, where the pretreated fractions are a cryoprecipitate liberated from fibrinogen, globulin, albumins, and other interfering ingredients by means of ethanol precipitation.
EP-A-0 343 275 describes chromatographic separation with ion exchangers after virus inactivation of the cryoprecipitate fraction. EP 0 173 242 A describes a method for obtaining factor VIII preparations by chromatography on anion-exchanging materials which are solely based on carbohydrates, the carbohydrate matrix being modified by DEAF groups. In particular, DEAF sepharose and DEAF cellulose are described as being useful. In GB-A-1,178,958, a purification of factor VIII
using ECTEOLA cellulose columns is described. The modified cellulose contains basic substituents introduced by reaction of epichlorohydrin and triethanolamine. The above mentioned prior art makes use either of chromatographic separation in the form of batch processes or of column chromatography.
Although fairly good results are obtained with these methods, it still remains desirable for economic as well as ethical reasons to increase the yield of biologically valuable factor VIII.
Hence, the technical problem underlying this invention consists in providing a method which, proceeding from the prior art, allows for an improved preparation of factor VIII
with respect to yield and biological activity.
Surprisingly, the problem has been solved by a process proceeding from a cryoprecipitate after dissolution thereof or blood plasma, optionally followed by an aluminum hydroxide treatment, and employing a separation step using membrane chromatography on membranes or compact disks of the porous hydrophilic polymers poly(glycidyl methacrylates) or hydrophilized polystyrene.
The process of the invention can be performed with commercial cryoprecipitate or blood plasma. Preferably, the thawed cryoprecipitate is treated with aluminium hydroxide for further pre-purification of the sample in order to preconcentrate factor VIII.
Preferably, a virus inactivation is carried out prior to the actual chromatographic purification on materials arranged within or on membranes. This virus inactivation is performed according to the method described in EP 131 740 A1 by treatment with biocompatible organic solvents (detergents), Triton~ X-100/TNBP, preferably Tween~/TNBP (tri-n-butyl phosphate). Good results will also be obtained with sodium cholate/TNPB. Preferably, detergents are employed in amounts of up to 15o by weight. Wherein a virus inactivation is performed by treatment of said fraction with a di- or trialkyl phosphate and a non-ionic surfactant.
The chromatographic separation step for the purification of factor VIII in the sample may be carried out either on base materials modified by ion-exchanging groups, particularly anion exchangers, or on materials modified by immunoaffinity ligands. It is critical that the mentioned materials are arranged in membranes. Particularly useful are membranes as well as compact disks made of porous poly(glycidyl methacrylates) and/or hydrophilized polystyrene.
A membrane suitable for separation consists of compact disks made of polymer carriers. The base materials of the membranes or disks are provided with corresponding anion-exchanging groups or immunoaffinity ligands. Particularly anion-exchanging groups such as quaternary amines or diethylaminoethyl groups are considered as ion-exchanging groups. Suitable ca n on exchangers are in principle weakly and strongly acidic ration exchangers, such as materials which are modified by sulfonic acid or phosphoric acid groups.
The ion-exchanging groups can be bound to the base material fibers with or without a so-called spacer.
Materials provided with spacers are also referred to as tentacle materials. In DE 42 04 694 corresponding spacers and ligands are mentioned. For example, a glucosamine residue can serve as a spacer as well. Anion-exchanging groups such as DEAF or quaternary amines may also be bound at the membranes made of porous poly(glycidyl methacrylate) or the other materials mentioned. The binding of the anion-exchanging groups takes place either directly to the material forming the membrane or else through a spacer as well, e.g. a glucosamine residue.
In another embodiment of the process of the invention affinity membrane chromatography with immobilized substances having a high affinity for factor VIII is used.
The substances having affinity for factor VIII are immobilized at the carrier by means of chemically reactive groups. Preferably, the reactive group will attack at the end of a spacer rather than directly at the carrier material. Immobilization of the substances for factor VIII
takes place through binding at reactive groups such as tosyl, tresyl, hydrazid and others. Corresponding procedures are known from T. M. Phillips "Affinity chromatography" in "Chromatography" (E. Heftmann, ed.), 5th ed. Elsevier, Amsterdam 1992.
In another preferred embodiment materials are employed for the separation of factor VIII that can ensure a hydrophobic interaction. Hydrophobic materials which are employed are acyclic and/or cyclic alkyl chains, for instance C1 to C18 alkyl chains, and aromatic substances.
Suitable materials providing hydrophobic interaction preferably include those having graded hydrophobicity.
Hydrophobicity can be graded by introduction of polar groups, such as protic-polar or aprotic-polar groups, for example hydroxy, amino, cyano groups. Preferably, it is adapted corresponding to the respective separation conditions.
Virus inactivation may also take place by heat treatment. Preferably, the eluated sample containing factor VIII is subjected to a pasteurization step following a first membrane chromatographic step. A corresponding procedure is proposed in DE-A-4 318 435. Therein, fraction enriched with factor VIII are contacted with di- or trialkyl phosphates and optionally wetting agents in the presence of stabilizing agents and simultaneously or subsequently treated for a period of 5 hours to 30 hours at elevated temperatures ranging from 55°C to 67°C. It may be of advantage to combine the two methods of virus inactivation, the treatment with detergents and with heat.
In order to remove the chemicals employed in the pasteurization step, a second membrane chromatography may follow. Preferably, the separation of the added stabilizing agents takes place by means of a membrane modified by DEAF or quaternary ammonium compounds that are arranged on the surface of the chromatographic carrier material through a spacer. In addition, it is possible to arrange the corresponding ligands on the surface of the carrier material without a spacer. The stabilizing agents are not retarded by this anion exchange material under the conditions chosen, whereas factor VIII is adsorbed on the chromatographic material.
Thereafter, factor VIII is eluated with an aqueous solvent system exhibiting gradually increasing salt concentrations.
The fraction containing factor VIII thus obtained is concentrated, filled and optionally lyophilized by conventional methods. Preferably, factor VIII is applied from a solution with low ionic strength in the first membrane chromatographic separation step.
Preferably, the aqueous system has an ionic strength corresponding to a 0 to 150 mM sodium chloride solution. At such ionic strengths, factor VIII is still adsorbed on the chromatographic material, whereas more weakly binding impurities can be washed out with aqueous systems of the same ionic strength.
In another embodiment of the process according to the invention, purification of the adsorbed material may be performed with an aqueous system having an ionic strength corresponding to a 200 to 400 mM sodium chloride solution.
Desorption of factor VIII and elution of this fraction then are carried out with an aqueous system having an ionic strength corresponding to a 500 to 1500 mm sodium chloride solution, while the pH value is maintained within a range of 4 to 9. If cation-exchange chromatography is performed, it preferably takes place at a pH value < 6, whereas anion-exchange chromatography is rather carried out at higher pH
values of above 6.
If purification of factor VIII is performed by immunoaffinity membrane chromatography, then, unlike with the above mentioned method using anion-exchange materials, elution is carried out with chaotropic reagents or highly concentrated salt solutions. Preferably, elution takes place with concentrations of chaotropic reagents or salts which are sufficient to disrupt the binding between the substance having high affinity to factor VIII and factor VIII itself.
The concentrations of the mentioned materials in the respective elution systems depends on the strength of the affinity between factor VIII and the corresponding binding component. As a result, elution can take place with aqueous solutions with lower denaturating potency. Preferably, aqueous solutions having concentrations of from 1 to 6 M of urea, especially from 2 to 4 M of urea, or correspondingly highly concentrated salt solutions are employed for elution of factor VIII from the immunoaffinity membrane.
In hydrophobic interaction chromatography, the sample is applied in an aqueous solution with very high ionic strength, such as, for example, highly concentrated ammonium sulfate (up to a concentration of 4 M) or sodium chloride (up to a concentration of up to 5 M). In particular, elution is carried out gradually or continuously with salt solutions of lower ionic strengths. An aqueous solution with organic solvents, particularly a diluted alcoholic solution, may also be used as solutions with lower ionic strengths for the elution of the samples in hydrophobic interaction membrane chromatography.
The procedure according to the invention surprisingly ensures a quick and uncomplicated purification of factor VIII which is obtained at the same time in high purity and high yields. In addition, the specific activity of the factor VIII obtained in this way is quite high which may be accounted for by the low denaturation of the active factor in the process according to the invention.
Preferably, the aqueous system has an ionic strength corresponding to a 0 to 150 mM sodium chloride solution. At such ionic strengths, factor VIII is still adsorbed on the chromatographic material, whereas more weakly binding impurities can be washed out with aqueous systems of the same ionic strength.
In another embodiment of the process according to the invention, purification of the adsorbed material may be performed with an aqueous system having an ionic strength corresponding to a 200 to 400 mM sodium chloride solution.
Desorption of factor VIII and elution of this fraction then are carried out with an aqueous system having an ionic strength corresponding to a 500 to 1500 mm sodium chloride solution, while the pH value is maintained within a range of 4 to 9. If cation-exchange chromatography is performed, it preferably takes place at a pH value < 6, whereas anion-exchange chromatography is rather carried out at higher pH
values of above 6.
If purification of factor VIII is performed by immunoaffinity membrane chromatography, then, unlike with the above mentioned method using anion-exchange materials, elution is carried out with chaotropic reagents or highly concentrated salt solutions. Preferably, elution takes place with concentrations of chaotropic reagents or salts which are sufficient to disrupt the binding between the substance having high affinity to factor VIII and factor VIII itself.
The concentrations of the mentioned materials in the respective elution systems depends on the strength of the affinity between factor VIII and the corresponding binding component. As a result, elution can take place with aqueous solutions with lower denaturating potency. Preferably, aqueous solutions having concentrations of from 1 to 6 M of urea, especially from 2 to 4 M of urea, or correspondingly highly concentrated salt solutions are employed for elution of factor VIII from the immunoaffinity membrane.
In hydrophobic interaction chromatography, the sample is applied in an aqueous solution with very high ionic strength, such as, for example, highly concentrated ammonium sulfate (up to a concentration of 4 M) or sodium chloride (up to a concentration of up to 5 M). In particular, elution is carried out gradually or continuously with salt solutions of lower ionic strengths. An aqueous solution with organic solvents, particularly a diluted alcoholic solution, may also be used as solutions with lower ionic strengths for the elution of the samples in hydrophobic interaction membrane chromatography.
The procedure according to the invention surprisingly ensures a quick and uncomplicated purification of factor VIII which is obtained at the same time in high purity and high yields. In addition, the specific activity of the factor VIII obtained in this way is quite high which may be accounted for by the low denaturation of the active factor in the process according to the invention.
Claims (14)
1. A process for the preparation of a virus inactivated fraction containing factor VIII by means of chromatographic methods, wherein - proceeding from a cryoprecipitate after dissolution thereof or blood plasma - followed by aluminium hydroxide treatment, a virus inactivation is performed by treatment of said fraction with a di- or trialkyl phosphate and a non-ionic surfactant, followed by at least one separation step using membrane chromatography on membranes or compact disks of the porous hydrophilic polymers poly(glycidyl methacrylates) or hydrophilized polystyrene.
2. The process according to claim 1, wherein said separation step takes place on an ion-exchange material, provided within or on a membrane.
3. The process according to claim 2, wherein the ion-exchange material is an anion-exchange material.
4. The process according to claim 1 or 2, wherein the virus inactivation is preceded by a pasteurization step.
5. The process according to claim 4, wherein the pasteurization step is followed by an additional separation step using membrane chromatography.
6. The process according to at least one of claims 1 or 4, wherein said membrane chromatography takes place on a material having a high affinity for factor VIII.
7. The process according to at least one of claims 1 or 6, wherein said material having a high affinity for factor VIII is modified by ligands having high or low molecular weights.
8. The process according to claim 6 or 7, wherein the modified material having a high affinity for factor VIII
bears immobilized ligands having a high affinity for factor VIII.
bears immobilized ligands having a high affinity for factor VIII.
9. The process according to any one of claims 1 to 5, wherein said chromatographic material allows for hydrophobic interaction with the factor VIII to be separated or bears appropriate ligands mediating hydrophobic interaction.
10. The process according to at least one of claims 1 to 5, wherein the sample to be purified is applied to the membrane containing ion-exchange material in an aqueous system having a low ionic strength, corresponding to 0 to 150 mM sodium chloride, optionally washed with an aqueous system having a higher ionic strength corresponding to a 200 to 400 mM sodium chloride solution, followed by elution with an aqueous system having a high ionic strength corresponding to a 500 to 1,500 mM sodium chloride solution, while the pH value is maintained at from 4 to 9.
11. The process according to claim 10, wherein the sample is washed with an aqueous system having higher ionic strength, corresponding to a 200 to 400 mM sodium chloride solution, followed by elution with an aqueous system having high ionic strength, corresponding to a 500 to 1,500 mM sodium chloride solution, while the pH value is maintained at from 4 to 9.
12. The process according to at least one of claims 1, 5 or 9, wherein the sample to be purified is applied from a solution having a very high ionic strength to a membrane bearing hydrophobic ligands on its surface, and is eluted with solvent systems having lower ionic strengths.
13. The process according to at least one of claims 1 to 11, wherein the eluted fraction containing factor VIII is concentrated, filled into appropriate containers or lyophilized.
14. The process according to at least one of claims 1 to 12, wherein said virus inactivation is performed by treatment with up to 15% by weight of a detergent.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE4337573A DE4337573C1 (en) | 1993-11-04 | 1993-11-04 | Process for the preparation of a virus-inactivated factor VIII-containing fraction by means of chromatographic methods |
| DEP4337573.1 | 1993-11-04 | ||
| PCT/EP1994/003258 WO1995012609A1 (en) | 1993-11-04 | 1994-09-30 | Process for producing a virus-inactivated factor viii-containing fraction by chromatographic methods |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2175670A1 CA2175670A1 (en) | 1995-05-11 |
| CA2175670C true CA2175670C (en) | 2005-08-02 |
Family
ID=6501733
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002175670A Expired - Fee Related CA2175670C (en) | 1993-11-04 | 1994-09-30 | Process for producing a virus-inactivated factor viii-containing fraction by chromatographic methods |
Country Status (21)
| Country | Link |
|---|---|
| EP (1) | EP0726907B1 (en) |
| JP (1) | JP3739788B2 (en) |
| KR (1) | KR960705841A (en) |
| CN (1) | CN1109045C (en) |
| AT (1) | ATE228533T1 (en) |
| AU (1) | AU687451B2 (en) |
| CA (1) | CA2175670C (en) |
| CZ (1) | CZ290186B6 (en) |
| DE (2) | DE4337573C1 (en) |
| ES (1) | ES2182846T3 (en) |
| FI (1) | FI116569B (en) |
| HU (1) | HU222087B1 (en) |
| IL (1) | IL111263A (en) |
| MY (1) | MY113294A (en) |
| NO (1) | NO317112B1 (en) |
| PL (1) | PL181725B1 (en) |
| RU (1) | RU2148411C1 (en) |
| SK (1) | SK284215B6 (en) |
| UA (1) | UA43855C2 (en) |
| WO (1) | WO1995012609A1 (en) |
| ZA (1) | ZA948667B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0885394B1 (en) * | 1996-03-08 | 2000-08-16 | Octapharma AG | Process for testing suitability of protein fractions containing factor viii |
| DE19618851C1 (en) * | 1996-05-10 | 1997-07-24 | Octapharma Ag | Testing factor VIII containing preparations |
| ES2137878B1 (en) * | 1997-12-01 | 2000-10-01 | Grifols Grupo Sa | PROCEDURE FOR OBTAINING AN ATTENUATED HUMAN PLASMA OF VIRUSES FOR THERAPEUTIC USE. |
| DK2300497T3 (en) | 2008-06-24 | 2012-10-29 | Octapharma Ag | Method of purifying coagulation factor VIII |
| AU2011343813B2 (en) | 2010-12-15 | 2015-05-21 | Takeda Pharmaceutical Company Limited | Eluate collection using conductivity gradient |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4540573A (en) * | 1983-07-14 | 1985-09-10 | New York Blood Center, Inc. | Undenatured virus-free biologically active protein derivatives |
| DE3432083A1 (en) * | 1984-08-31 | 1986-03-06 | Behringwerke Ag, 3550 Marburg | PASTEURIZED, ISOAGGLUTININ-FREE FACTOR VIII PREPARATION AND METHOD FOR THE PRODUCTION THEREOF |
| US5043428A (en) * | 1984-08-31 | 1991-08-27 | Behringwerke Aktiengesellschaft | Pasteurized, isoagglutinin-free factor VIII preparation and a process for its production |
| JPS62191042A (en) * | 1986-02-17 | 1987-08-21 | Kanegafuchi Chem Ind Co Ltd | Blood coagulation factor viii adsorbent and method for purifying blood coagulation factor viii using same |
| CA1339946C (en) * | 1987-03-31 | 1998-07-07 | Michael J. Griffith | Ultrapurification process for polypeptides |
| US4774323A (en) * | 1987-06-29 | 1988-09-27 | Rorer Pharmaceutical Corporation | Purification of von Willebrand Factor solutions using gel permeation chromatography |
| US4795806A (en) * | 1987-07-16 | 1989-01-03 | Miles Laboratories, Inc. | Phospholipid affinity purification of Factor VIII:C |
| CA2001720C (en) * | 1988-10-31 | 2001-10-02 | Randal A. Goffe | Membrane affinity apparatus and purification methods related thereto |
| DE3878245D1 (en) * | 1988-11-05 | 1993-03-18 | Octapharma Ag | METHOD FOR PRODUCING A HIGHLY PURE, NON-INFECTIOUS ANTIHAEMOPHILIE ACTUATOR BY CHROMATOGRAPHY. |
| CH678155A5 (en) * | 1989-08-09 | 1991-08-15 | Fischer Ag Georg | |
| AT399818B (en) * | 1992-04-24 | 1995-07-25 | Immuno Ag | METHOD FOR PRODUCING A HIGH PURIFIED VIRUS-SAFE FACTOR VIII PREPARATION |
-
1993
- 1993-11-04 DE DE4337573A patent/DE4337573C1/en not_active Revoked
-
1994
- 1994-09-30 HU HU9601192A patent/HU222087B1/en not_active IP Right Cessation
- 1994-09-30 WO PCT/EP1994/003258 patent/WO1995012609A1/en active IP Right Grant
- 1994-09-30 SK SK566-96A patent/SK284215B6/en not_active IP Right Cessation
- 1994-09-30 JP JP51297795A patent/JP3739788B2/en not_active Expired - Fee Related
- 1994-09-30 AU AU78109/94A patent/AU687451B2/en not_active Expired
- 1994-09-30 CZ CZ19961187A patent/CZ290186B6/en not_active IP Right Cessation
- 1994-09-30 RU RU96110890A patent/RU2148411C1/en not_active IP Right Cessation
- 1994-09-30 PL PL94314185A patent/PL181725B1/en not_active IP Right Cessation
- 1994-09-30 KR KR1019960702337A patent/KR960705841A/en not_active Application Discontinuation
- 1994-09-30 EP EP94928846A patent/EP0726907B1/en not_active Expired - Lifetime
- 1994-09-30 DE DE59410213T patent/DE59410213D1/en not_active Expired - Lifetime
- 1994-09-30 CN CN94194020A patent/CN1109045C/en not_active Expired - Fee Related
- 1994-09-30 CA CA002175670A patent/CA2175670C/en not_active Expired - Fee Related
- 1994-09-30 AT AT94928846T patent/ATE228533T1/en active
- 1994-09-30 UA UA96041756A patent/UA43855C2/en unknown
- 1994-09-30 ES ES94928846T patent/ES2182846T3/en not_active Expired - Lifetime
- 1994-10-12 IL IL11126394A patent/IL111263A/en not_active IP Right Cessation
- 1994-11-03 ZA ZA948667A patent/ZA948667B/en unknown
- 1994-11-03 MY MYPI94002921A patent/MY113294A/en unknown
-
1996
- 1996-05-03 FI FI961900A patent/FI116569B/en active IP Right Grant
- 1996-05-03 NO NO19961816A patent/NO317112B1/en not_active IP Right Cessation
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Legal Events
| Date | Code | Title | Description |
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| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20140930 |