US4985544A
Expired / Lapsed
US
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Process for renaturing fish growth hormone
Assignee: KH Neochem Co Ltd
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Filing: 1988-08-02
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Granted: 1991-01-15
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Expires: 2008-08-02
Abstract
Abstract Reactivation of cysteine-containing protein in a process, in which a reduced and denatured cysteine-containing protein such as salmon growth hormone I or eel growth hormone I can be efficiently reactivated.
Inventors
- 👤 Yoshiharu Yokoo
- 👤 Seiji Sugimoto
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 01:32
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/107; C07K1/113; C07K1/1133; C07K14/00; C07K14/435; C07K14/575; C07K14/61
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
90 words
A process for renaturing a fish growth hormone polypeptide in its natural form which consists of the successive steps of: (1) converting a fish growth hormone polypeptide into the corresponding reduced and denatured protein by adding a denaturing agent and a reducing agent thereto to solubilize the protein; (2) removing the reducing agent; (3) oxidizing the protein, as denatured, to form disulfide bonds at the same sites as those observed in the corresponding natural protein; and thereafter (4) removing the denaturing agent to thereby isolate and purify the renatured protein.
Claim 2
Dependent Claim
90 words
A process for renaturing a fish growth hormone polypeptide in its natural form which consists of the successive steps of: (1) converting a fish growth hormone polypeptide into the corresponding reduced and denatured protein by adding a denaturing agent and a reducing agent thereto to solubilize the protein; (2) removing the reducing agent; (3) oxidizing the protein, as denatured, to form disulfide bonds at the same sites as those observed in the corresponding natural protein; and thereafter (4) removing the denaturing agent to thereby isolate and purify the renatured protein.
Claim 3
Dependent Claim
39 words
A process of renaturing a fish growth hormone polypeptide in its natural form from inclusion bodies produced within the cells of a microorganism, said process consisting of the successive steps of: fractionation range (molecular weight) is 5000-250000 for proteins.
Claim 4
Dependent Claim
39 words
A process of renaturing a fish growth hormone polypeptide in its natural form from inclusion bodies produced within the cells of a microorganism, said process consisting of the successive steps of: fractionation range (molecular weight) is 5000-250000 for proteins.
Claim 5
Dependent Claim
28 words
A process as set forth in claim 1 or 2, wherein said denaturing agent is selected from the group consisting of SDS, urea, guanidine hydrochloride, acids and alkalis.
Claim 6
Dependent Claim
40 words
A process as set forth in claim 1 or 2, wherein the reducing agent is removed in the presence of a denaturing agent under non-oxidizing conditions by dilution, dialysis, ultrafiltration, gel filtration, adsorption/desorption chromatography or a batch process of adsorption/desorption.
Claim 7
Dependent Claim
29 words
A process as set forth in claim 1 or 2, wherein the reducing agent is removed by gel filtration in the presence of a denaturing agent under non-oxidizing conditions.
Claim 8
Dependent Claim
16 words
A process as set forth in claim 1 or 2, wherein the oxidation process is spontaneous.
Claim 9
Dependent Claim
49 words
A process as set forth in claim 1 or 2, wherein said oxidation is carried out by a method selected from the group consisting of air-oxidation with aeration, air-oxidation with the use of a metal ion as a catalyst, and oxidation with the use of a weak oxidizing agent.
Claim 10
Dependent Claim
40 words
A process as set forth in claim 7, wherein said weak oxidizing agent is selected from the group consisting of o-iodosobenzoic acid, oxidized glutathione, a mixture of oxidized glutathione and reducing glutathione, cystine or a mixture of cystine and cysteine.
Claim 11
Dependent Claim
26 words
A process as set forth in claim 1 or 2, wherein the protein concentration during the oxidation step is from about 0.1 to about 2,000 μg/ml.
Claim 12
Dependent Claim
20 words
A process as set forth in claim 1 or 2, in which the cysteine-containing protein is salmon growth hormone I.
Claim 13
Dependent Claim
20 words
A process as set forth in claim 1 or 2, in which the cysteine-containing protein is eel growth hormone I.
References Cited (Backward Citations)
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EP0166444A2
Cited
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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) | 1994-01-14 | 1994-07-16 | 1995-01-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-01-14 | 1998-07-16 | 1999-01-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-01-14 | 2002-07-16 | 2003-01-15 | $7700.00 | Window Closed / Lapsed |