US4122250A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Separation of high-activity heparin by affinity chromatography
Assignee: Individual
•
Filing: 1977-08-22
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Granted: 1978-10-24
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Expires: 1997-08-22
Abstract
Abstract An affinity chromatography method for the separation of high-activity heparin from a mixture thereof with low-activity heparin. The method employs an affinity column of DEAE-Sephadex and a series of sodium chloride-imidazole elution buffers (pH 6.5-7.5) varying in sodium chloride molarity from about 0.5 to about 1.0. Stepwise elution of a plurality of heparin fractions from the affinity column with the elution buffers of successively increasing sodium chloride molarity results in the removal from the affinity column of first the low-activity heparin and finally the high-activity heparin.
Inventors
- 👤 Gottfried Schmer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:29
CPC Classifications
C; C08; C08B; C08B37/00; C08B37/006; C08B37/0063; C08B37/0075
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
153 words
An affinity chromatography method for the separation of heparin of relatively high specific activity from a mixture thereof with heparin of relatively low specific activity, comprising the steps of: (a) preparing a solution of said heparin mixture in a solvent of about 0.5 molar sodium chloride buffered with about 0.01-0.03 molar imidazole to pH 6.5-7.5; (b) applying said heparin solution onto an affinity column of packed gel comprising the diethylaminoethyl ether of cross-linked polydextran; and (c) successively eluting a plurality of heparin fractions from said affinity column with a series of elution buffers of successively increasing molarity of sodium chloride buffered with about 0.01-0.03 molar imidazole to pH 6.5-7.5, said elution buffers having a sodium chloride molarity ranging from about 0.5 to about 1.0, so as to effect a stepwise removal from said affinity column of first the heparin of relatively low specific activity and finally the heparin of relatively high specific activity.
Claim 2
Dependent Claim
153 words
An affinity chromatography method for the separation of heparin of relatively high specific activity from a mixture thereof with heparin of relatively low specific activity, comprising the steps of: (a) preparing a solution of said heparin mixture in a solvent of about 0.5 molar sodium chloride buffered with about 0.01-0.03 molar imidazole to pH 6.5-7.5; (b) applying said heparin solution onto an affinity column of packed gel comprising the diethylaminoethyl ether of cross-linked polydextran; and (c) successively eluting a plurality of heparin fractions from said affinity column with a series of elution buffers of successively increasing molarity of sodium chloride buffered with about 0.01-0.03 molar imidazole to pH 6.5-7.5, said elution buffers having a sodium chloride molarity ranging from about 0.5 to about 1.0, so as to effect a stepwise removal from said affinity column of first the heparin of relatively low specific activity and finally the heparin of relatively high specific activity.
Claim 3
Dependent Claim
29 words
The method of claim 1, wherein the elution buffer employed in eluting the final heparin fraction of relatively high specific activity has a sodium chloride molarity of about 1.0.
Claim 4
Dependent Claim
24 words
The method of claim 2, wherein the elution buffer employed in eluting the penultimate heparin fraction has a sodium chloride molarity of about 0.75.
Claim 5
Dependent Claim
49 words
The method of claim 3, wherein said series of elution buffers comprises a first elution buffer of 0.5 molar sodium chloride, a second elution buffer of 0.6 molar sodium chloride, a third elution buffer of 0.75 molar sodium chloride, and a fourth elution buffer of 1.0 molar sodium chloride.
Claim 6
Dependent Claim
23 words
The method of claim 4, wherein said solvent and each of said elution buffers is buffered with 0.02 molar imidazole to pH 7.35.
Claim 7
Dependent Claim
25 words
The method of claim 1, wherein said affinity column is equilibrated with an additional amount of said solvent prior to application of said heparin solution.
Claim 8
Dependent Claim
33 words
The method of claim 1, wherein the amount of said heparin solution which is applied onto said affinity column is equivalent to about 2 grams of heparin per 100 ml of packed gel.
Claim 9
Dependent Claim
29 words
The method of claim 1, wherein the amount of elution buffer which is employed for eluting each heparin fraction is about 200-400 ml per 100 ml of packed gel.
References Cited (Backward Citations)
US3099600A
Cited
DE1195010B
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-10-23 | 1982-04-24 | 1982-10-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-10-23 | 1986-04-24 | 1986-10-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-10-23 | 1990-04-24 | 1990-10-24 | $7700.00 | Window Closed / Lapsed |