US4499014A
Expired / Lapsed
US
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Method for purifying gamma-interferon
Assignee: Interferon Sciences Inc
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Filing: 1984-02-07
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Granted: 1985-02-12
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Expires: 2004-02-07
Abstract
Abstract A method is provided for restoring some or all of the activity of gamma-interferon which has been in contact with an acidic solution comprising the steps of: (a) placing the gamma-interferon in a solution which has a pH between about 5.5 and 9.5; and (b) incubating the solution at a temperature of between about 2° C. and 8° C. for a period of at least 24 hours. The method allows antibody affinity chromatography employing acid elution to be used to purify gamma-interferon, in that, the activity of the acid-eluted gamma-interferon can be essentially completely restored using the reactivation process.
Inventors
- 👤 Leonard F. Estis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 12:28
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/18; C07K16/24; C07K16/249; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/57; Y; Y10; Y10S; Y10S435/00; Y10S435/811; Y10S530/00; Y10S530/808; Y10S530/809; Y10S530/827; Y10S530/829
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
185 words
A method for purifying gamma-interferon comprising the steps of: (a) preparing one or more antibodies to gamma-interferon; (b) immobilizing the one or more antibodies on a solid support; (c) contacting a first solution containing gamma-interferon with the one or more immobilized antibodies so as to form immobilized antibody-antigen complexes between the gamma-interferon and the one or more immobilized antibodies; (d) separating the first solution from the immobilized antibody-antigen complexes; (e) contacting the immobilized antibody-antigen complexes with a second solution having an acidic pH so as to disassociate the gamma-interferon from the one or more immobilized antibodies and into the second solution, the acidity of said second solution having the effect of partially or completely deactivating the diassociated gamma-interferon; (f) separating the second solution from the one or more immobilized antibodies; and (g) restoring some or all of the activity of the gamma-interferon in the second solution by adjusting the pH of that solution to between about 5.5 and 9.5 and then incubating the pH adjusted solution at a temperature of between about 2° C. and 8° C. for a period of at least 24 hours.
Claim 2
Independent Claim
185 words
A method for purifying gamma-interferon comprising the steps of: (a) preparing one or more antibodies to gamma-interferon; (b) immobilizing the one or more antibodies on a solid support; (c) contacting a first solution containing gamma-interferon with the one or more immobilized antibodies so as to form immobilized antibody-antigen complexes between the gamma-interferon and the one or more immobilized antibodies; (d) separating the first solution from the immobilized antibody-antigen complexes; (e) contacting the immobilized antibody-antigen complexes with a second solution having an acidic pH so as to disassociate the gamma-interferon from the one or more immobilized antibodies and into the second solution, the acidity of said second solution having the effect of partially or completely deactivating the diassociated gamma-interferon; (f) separating the second solution from the one or more immobilized antibodies; and (g) restoring some or all of the activity of the gamma-interferon in the second solution by adjusting the pH of that solution to between about 5.5 and 9.5 and then incubating the pH adjusted solution at a temperature of between about 2° C. and 8° C. for a period of at least 24 hours.
Claim 3
Dependent Claim
19 words
The method of claim 1 wherein the acidic pH of the second solution is between about 1.5 and 4.0.
Claim 4
Dependent Claim
20 words
The method of claim 1 wherein the pH of the second solution is adjusted to between about 6.0 and 9.0.
Claim 5
Dependent Claim
20 words
The method of claim 1 wherein the pH adjusted second solution is incubated at a temperature of about 4° C.
Claim 6
Dependent Claim
23 words
The method of claim 1 wherein the pH adjusted second solution is stored for a period of between about 24 and 96 hours.
Claim 7
Dependent Claim
20 words
The method of claim 5 wherein the pH adjusted second solution is stored for a period of about 96 hours.
Claim 8
Dependent Claim
37 words
The method of claim 1 wherein the pH of the second solution is adjusted to between about 6.0 and 9.0 and the solution is then stored at a temperature of about 4° C. for about 96 hours.
Claim 9
Independent Claim
65 words
A method for restoring some or all of the activity of gamma-interferon which has been in contact with an acidic solution comprising the steps of: (a) placing the gamma-interferon in a solution which has a pH between about 5.5 and 9.5; and (b) incubating the solution at a temperature of between about 2° C. and 8° C. for a period of at least 24 hours.
Claim 10
Independent Claim
65 words
A method for restoring some or all of the activity of gamma-interferon which has been in contact with an acidic solution comprising the steps of: (a) placing the gamma-interferon in a solution which has a pH between about 5.5 and 9.5; and (b) incubating the solution at a temperature of between about 2° C. and 8° C. for a period of at least 24 hours.
Claim 11
Dependent Claim
17 words
The method of claim 8 wherein the pH of the solution is between about 6.0 and 9.0.
Claim 12
Dependent Claim
17 words
The method of claim 8 wherein the solution is incubated at a temperature of about 4° C.
Claim 13
Dependent Claim
20 words
The method of claim 8 wherein the solution is incubated for a period of between about 24 and 96 hours.
Claim 14
Dependent Claim
17 words
The method of claim 11 wherein the solution is incubated for a period of about 96 hours.
Claim 15
Dependent Claim
35 words
The method of claim 8 wherein the pH of the solution is between about 2° C. and 8° C. and the solution is incubated at a temperature of about 4° C. for about 96 hours.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-02-12 | 1988-08-13 | 1989-02-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-02-12 | 1992-08-13 | 1993-02-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-02-12 | 1996-08-13 | 1997-02-12 | $7700.00 | Window Closed / Lapsed |