US5310880A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Purification of Mullerian inhibiting substance
Assignee: General Hospital Corp
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Filing: 1991-04-12
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Granted: 1994-05-10
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Expires: 2011-04-12
Abstract
Abstract The present invention discloses a method of purifying Mullerian Inhibiting Substance. The method takes advantage of immunoaffinity chromatography to achieve MIS at up to 95% purity. The method provides for a purified MIS product in which contaminating enzymes having MIS proteolytic activity or inhibitors of MIS antiproliferative activity are substantially removed by addition of an alkali metal halide solution or an alkaline earth metal halide solution prior to elution with an acid solution having a pH of between about 2.5 and 4.0.
Inventors
- 👤 Patricia K. Donahoe
- 👤 Richard C. Ragin
- 👤 David T. MacLaughkin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 22:31
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/575; A; A61; A61K; A61K38/00
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
90 words
A method of purifying recombinant MIS from host cells capable of expressing recombinant MIS comprising (a) binding the recombinant MIS to an antibody-chromatography matrix, said antibody being specific to MIS, (b) substantially removing contaminating enzymes having MIS proteolytic activity or inhibitors of MIS antiproliferative activity by adding to the matrix an effective amount of an alkali metal halide solution wherein said solution contains an effective amount of chelating agent, and (c) recovering the recombinant MIS by eluting with an acid solution having a pH of between about 2.5 and 4.0.
Claim 2
Independent Claim
90 words
A method of purifying recombinant MIS from host cells capable of expressing recombinant MIS comprising (a) binding the recombinant MIS to an antibody-chromatography matrix, said antibody being specific to MIS, (b) substantially removing contaminating enzymes having MIS proteolytic activity or inhibitors of MIS antiproliferative activity by adding to the matrix an effective amount of an alkali metal halide solution wherein said solution contains an effective amount of chelating agent, and (c) recovering the recombinant MIS by eluting with an acid solution having a pH of between about 2.5 and 4.0.
Claim 3
Dependent Claim
20 words
The method of claim 1, further comprising neutralizing the recovered MIS to a pH of between about 6.8 and 7.6.
Claim 4
Dependent Claim
18 words
The method of claim 1, wherein the alkali metal halide solution is between about 0.1M and 2.0 M.
Claim 5
Dependent Claim
13 words
The method of claim 1, wherein the alkali metal halide is sodium chloride.
Claim 6
Dependent Claim
11 words
The method of claim 1, wherein the acid is acetic acid.
Claim 7
Dependent Claim
17 words
The method of claim 3, wherein the alkali metal halide solution is between about 0.5M and 2.0M.
Claim 8
Dependent Claim
14 words
The method of claim 6, wherein the alkali metal halide solution is about 0.5M.
Claim 9
Dependent Claim
14 words
The method of claim 1, wherein said host cells are Chinese hamster ovary cells.
Claim 10
Dependent Claim
11 words
The method of claim 1, wherein said chelating agent is EDTA.
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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) | 1997-05-09 | 1997-11-08 | 1998-05-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-05-09 | 2001-11-08 | 2002-05-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-05-09 | 2005-11-08 | 2006-05-10 | $7700.00 | Window Closed / Lapsed |