Patent US3919044A successfully refreshed from Google Patents.
US3919044A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Processes for concentrating and purifying viruses and viral antigens
Assignee: Armour Pharmaceutical Co
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Filing: 1973-03-28
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Granted: 1975-11-11
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Expires: 1993-03-28
Abstract
Abstract Viruses and viral antigens harvested from embryonated eggs or tissue culture, whether acid-sensitive or acid-resistant, are purified and concentrated by processes which substantially eliminate the nonviral proteins present in the virus stock and thereafter permit the collection of a solute containing substantially all of the virus and viral antigens which were present in the starting stock. For both acid-resistant and acidsensitive viruses the processes of the invention comprise the elimination of nonviral protein by filtration or batch adsorption with resins discovered to selectively adsorb certain nonviral proteins. In the case of acid-sensitive viruses, the viruses are stabilized against subsequent acidification by the treatment of the resulting filtrate with a salt selected from the group consisting of ammonium sulfate, sodium sulfate, ammonium chloride and magnesium sulfate to complete the removal of nonviral protein. In both cases the viruses and viral antigens are acidified to pH 3-6 and adsorbed on preselected reactive materials from which the purified and concentrated virus is selectively extracted by elution with an eluent of pH 10-12 or higher which does not impair viral activity.
Inventors
- 👤 Joseph L Melnick
- 👤 Craig Wallis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 19:52
CPC Classifications
C; C12; C12N; C12N7/00; C12N2760/00; C12N2760/00011; C12N2760/16011; C12N2760/16111; C12N2760/16134; C12N2760/16151
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
237 words
A METHOD OF PURIFYING AND CONCENTRATING ACID-RESISTANT VIRUSES AND VIRAL ANTIGENS FOR VACCINATION FROM AN INITIAL STOCK SOLUTION CONTAINING ACID-RESISTANT VIRUS AND VIRAL ANTIGENS AND NON-VIRAL PROTEINS WHICH, UPON VACCINATION WILL SENSITIZE AND OTHERWISE CAUSE UNTOWARD ALLERGIC REACTIONS, SAID METHOD COMPRISING: A. TREATING SAID STOCK SOLUTION WITH 0.01 PERCENT ALKYLDIMETHYLBENZYL AMMONIUM CHLORIDE TO PRECIPITATE FROM SAID SOLUTION SAID NON-VIRAL PROTEINS WHICH FALL OUT OF SOLUTION IN AN ACID PH RANGE, B. FILTERING SAID TREATED SOLUTION THROUGH A COURSE CLARIFIER TO REMOVE THE PRECIPITATED PROTEINS THEREFROM, C. ADJUSTING THE PH OF SAID CLARIFIED SOLUTION TO 8-9 TO PREVENT VIRUS FROM ADSORBING TO RESINS, D. FILTERING SAID PH 8-9 CLARIFIED SOLUTION THR A CATIONIC RESIN EXCHANGER TO REMOVE ANY REMAINING NON-VIRAL PROTEINS FROM SAID SOLUTION AND PROVIDE A RESIN FILTRATE, E. ADJUSTING THE PH OF SAID RESIN FILTRATE TO 3-6 TO ENHANCE VIRUS ADSORPTION TO A MEMBRANE, F. FILTERING SAID ACIDIFIED RESIN FILTRATE THROUGH A 142 MM 0.45 MICRON REACTIVE MEMBRANE, FORMED OF A MATERIAL SELECTED FROM THE GROUP CONSISTING OF NITROCELLULOSE, EPOXY-FIBERGLASS, ASBESTOS, CELLULOSE, TRIACETATE POLYMERS, FIBERGLASS, OR REACTIVE TEXTILES, TO CONCENTRATE THE VIRUS ON SAID MEMBRANE, G. WASHING SAID MEMBRANE WITH A BUFFER HAVING A PH 3-9 TO REMOVE ALL RESIDUAL NON-VIRAL PROTEINS THEREFROM, H. TREATING SAID WASHED MEMBRANE WITH GLYCINE ELUENT AT PH 10-12 TO ELUTE THE VIRUS FROM SAID MEMBRANE, AND I. COLLECTING SAID ELUATE WHICH CONSISTS OF PURIFIED AND CONCENTRATED VIRUS AND VIRAL ANTIGENS.
Claim 2
Independent Claim
237 words
A METHOD OF PURIFYING AND CONCENTRATING ACID-RESISTANT VIRUSES AND VIRAL ANTIGENS FOR VACCINATION FROM AN INITIAL STOCK SOLUTION CONTAINING ACID-RESISTANT VIRUS AND VIRAL ANTIGENS AND NON-VIRAL PROTEINS WHICH, UPON VACCINATION WILL SENSITIZE AND OTHERWISE CAUSE UNTOWARD ALLERGIC REACTIONS, SAID METHOD COMPRISING: A. TREATING SAID STOCK SOLUTION WITH 0.01 PERCENT ALKYLDIMETHYLBENZYL AMMONIUM CHLORIDE TO PRECIPITATE FROM SAID SOLUTION SAID NON-VIRAL PROTEINS WHICH FALL OUT OF SOLUTION IN AN ACID PH RANGE, B. FILTERING SAID TREATED SOLUTION THROUGH A COURSE CLARIFIER TO REMOVE THE PRECIPITATED PROTEINS THEREFROM, C. ADJUSTING THE PH OF SAID CLARIFIED SOLUTION TO 8-9 TO PREVENT VIRUS FROM ADSORBING TO RESINS, D. FILTERING SAID PH 8-9 CLARIFIED SOLUTION THR A CATIONIC RESIN EXCHANGER TO REMOVE ANY REMAINING NON-VIRAL PROTEINS FROM SAID SOLUTION AND PROVIDE A RESIN FILTRATE, E. ADJUSTING THE PH OF SAID RESIN FILTRATE TO 3-6 TO ENHANCE VIRUS ADSORPTION TO A MEMBRANE, F. FILTERING SAID ACIDIFIED RESIN FILTRATE THROUGH A 142 MM 0.45 MICRON REACTIVE MEMBRANE, FORMED OF A MATERIAL SELECTED FROM THE GROUP CONSISTING OF NITROCELLULOSE, EPOXY-FIBERGLASS, ASBESTOS, CELLULOSE, TRIACETATE POLYMERS, FIBERGLASS, OR REACTIVE TEXTILES, TO CONCENTRATE THE VIRUS ON SAID MEMBRANE, G. WASHING SAID MEMBRANE WITH A BUFFER HAVING A PH 3-9 TO REMOVE ALL RESIDUAL NON-VIRAL PROTEINS THEREFROM, H. TREATING SAID WASHED MEMBRANE WITH GLYCINE ELUENT AT PH 10-12 TO ELUTE THE VIRUS FROM SAID MEMBRANE, AND I. COLLECTING SAID ELUATE WHICH CONSISTS OF PURIFIED AND CONCENTRATED VIRUS AND VIRAL ANTIGENS.
Claim 3
Dependent Claim
19 words
The method of claim 1 in which step (e) comprises adjusting the pH of said resin filtrate to 4-5.
Claim 4
Dependent Claim
21 words
The method of claim 1 in which step (g) comprises washing said membrane with a buffer having a pH of 4.5.
Claim 5
Dependent Claim
24 words
The method of claim 1 in which step (h) comprises treating said membrane with an eluent consisting of M glycine-NaOH buffer at pH 11.5.
Claim 6
Dependent Claim
93 words
The method claim 1 in which said virus and viral antigens are selected from the group consisting of influenza, measles, dengue, mumps, rubella, C-type viruses, cytomegalo-virus, vaccinia, canine distemper, canine hepatitis, infectious bovine rhinotracheitis, bovine virus diarrhea, bovine parainfluenza, Newcastle disease, Avian bronchitis, fowl pox, infectious laryngotracheitis, Marek''s disease, rabies virus, poliomyelitis variola, herpes virus, respiratory synctial, echoviruses 1, 7 and 9, coxsackie viruses A9, B1-6and canine hepatitis, said viruses known to be acid resistant being processed ''''as is'''' and those viruses known to be acid sensitive being rendered acid-resistant and then processed.
References Cited (Backward Citations)
US3078215A
Cited
US3655509A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-11-10 | 1979-05-12 | 1979-11-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-11-10 | 1983-05-12 | 1983-11-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-11-10 | 1987-05-12 | 1987-11-11 | $7700.00 | Window Closed / Lapsed |