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US4600531A
Expired / Lapsed
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Production of alpha-alpha cross-linked hemoglobins in high yield
Assignee: University of Iowa Research Foundation
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Filing: 1985-03-18
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Granted: 1986-07-15
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Expires: 2005-03-18
Abstract
Abstract A high yield method of production of a cross-linked hemoglobin derivative suitable for use as a blood substitute and plasma expander which is cross-linked between Lys 99 Alpha1 and Lys 99 Alpha2 in high percentage by adding the cross-linker in the presence of an added polyanion which blocks competing reactions at other sites of the protein.
Inventors
- 👤 Joseph A. Walder
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 06:44
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/795; C07K14/805; A; A61; A61P; A61P7/00; A61P7/08; A61K; A61K38/00
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
77 words
A method of producing in commercially significant yields, a modified hemoglobin which is capable of functioning as a blood substitute and blood plasma expander, comprising: isolating unmodified hemoglobin or preparing a crude red cell lysate; deoxygenating said hemoglobin; cross-linking said deoxygenated hemoglobin intramolecularly between Lys 99 Alpha 1 and Lys 99 Alpha 2 in the presence of an added polyanion selected from the group consisting of 2,3-diphosphoglycerate, inositol hexaphosphate, inositol hexasulfate; and purifying said alpha-alpha cross-linked hemoglobin.
Claim 2
Independent Claim
77 words
A method of producing in commercially significant yields, a modified hemoglobin which is capable of functioning as a blood substitute and blood plasma expander, comprising: isolating unmodified hemoglobin or preparing a crude red cell lysate; deoxygenating said hemoglobin; cross-linking said deoxygenated hemoglobin intramolecularly between Lys 99 Alpha 1 and Lys 99 Alpha 2 in the presence of an added polyanion selected from the group consisting of 2,3-diphosphoglycerate, inositol hexaphosphate, inositol hexasulfate; and purifying said alpha-alpha cross-linked hemoglobin.
Claim 3
Dependent Claim
10 words
The method of claim 1 wherein said polyanion is 2,3-diphosphoglycerate.
Claim 4
Dependent Claim
11 words
The method of claim 1 wherein said polyanion is inositol hexasulfate.
Claim 5
Dependent Claim
11 words
The method of claim 1 wherein said polyanion is inositol hexaphosphate.
Claim 6
Dependent Claim
22 words
The method of claim 1 wherein the reaction is conducted at a temperature of from about 0° C. to about 40° C.
Claim 7
Dependent Claim
22 words
The method of claim 5 wherein the reaction is conducted at a temperature of from about 35° C. to about 40° C.
Claim 8
Dependent Claim
17 words
The method of claim 6 wherein the reaction is conducted at a temperature of about 37° C.
Claim 9
Dependent Claim
27 words
The method of claim 1 wherein the cross-linking between Lys 99 Alpha 1 and Lys 99 Alpha 2 is with a cross-linking agent which is amino group-specific.
Claim 10
Dependent Claim
15 words
The method of claim 8 wherein said cross-linking agent is a phenyl ester cross-linking agent.
Claim 11
Dependent Claim
11 words
The method of claim 9 wherein said cross-linking agent is bis(3,5-dibromosalicyl)fumarate.
Claim 12
Dependent Claim
25 words
The method of claim 8 wherein the amount of cross-linking agent employed is up to a three fold molar excess of the amount of hemoglobin.
Claim 13
Dependent Claim
26 words
The method of claim 11 wherein the amount of cross-linking agent is from about 1.3 times to about 2.0 times the molar amount of said hemoglobin.
Claim 14
Dependent Claim
31 words
The method of claim 11 wherein the amount of the polyanion is from an amount equimolar to the amount of hemoglobin up to a twenty fold excess of the equimolar amount.
Claim 15
Dependent Claim
25 words
The method of claim 13 wherein the amount of the polyanion is from about 5 to about 10 times the molar amount of said hemoglobin.
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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) | 1989-07-14 | 1990-01-13 | 1990-07-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-07-14 | 1994-01-13 | 1994-07-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-07-14 | 1998-01-13 | 1998-07-15 | $7700.00 | Window Closed / Lapsed |