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US4508833A
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Separation of interleukin-2 from phytohemagglutinin by dye matrix chromatography
Assignee: Biotest AG
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Filing: 1982-12-14
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Granted: 1985-04-02
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Expires: 2002-12-14
Abstract
Abstract Crude interleukin-2 extract is subjected to group-selective dye-ligand absorption chromatography in one or more stages of purification with a matrix-gel medium consisting of a Blue A ligand or variant thereof or of a Green A ligand in a concentration of approximately 1.5 to 3.0 mg/ml of expanded matrix at a pH of approximately 6.8 to 8.5, a temperature of approximately 4° to 40° C., and a flowthrough rate of approximately 10 to 100 ml/h, employing an eluent. Either PHA-free or extremely pure interleukin-2 is obtained, depending on the overall number of purification stages.
Inventors
- 👤 Hans H. Sonneborn
- 👤 Udo Schwulera
- 👤 Hans Schleussner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 10:36
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6863; G01N33/6869; C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K14/55; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S436/00; Y10S436/807
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
83 words
A method of purifying human interleukin- 2 comprising passing crude interleukin-2 extract through a group-selective dye-ligand adsorption chromatographic column containing a matrix-gel medium consisting of a Blue A ligand or variant thereof or of a Green A ligand in a concentration of approximately 1.5 to 3.0 mg/ml of expanded matrix at a pH of approximately 6.8 to 8.5, a temperature of approximately 4° to 40° C., and a flowthrough rate of approximately 10 to 100 ml/h, and eluting purified interleukin-2 from the medium.
Claim 2
Independent Claim
83 words
A method of purifying human interleukin- 2 comprising passing crude interleukin-2 extract through a group-selective dye-ligand adsorption chromatographic column containing a matrix-gel medium consisting of a Blue A ligand or variant thereof or of a Green A ligand in a concentration of approximately 1.5 to 3.0 mg/ml of expanded matrix at a pH of approximately 6.8 to 8.5, a temperature of approximately 4° to 40° C., and a flowthrough rate of approximately 10 to 100 ml/h, and eluting purified interleukin-2 from the medium.
Claim 3
Dependent Claim
13 words
A method according to claim 1, wherein the ligand is Blue A ligand.
Claim 4
Dependent Claim
14 words
A method according to claim 1, wherein the matrix gel medium comprises 5% agarose.
Claim 5
Dependent Claim
14 words
A method according to claim 1, wherein elution is effected with NaCl or KCl.
Claim 6
Dependent Claim
41 words
A method according to claim 1, wherein prior to passage through the column the crude interleukin-2 is contacted with solutions of different concentrations of ammonium sulfate to effect selective precipitation in stages and the desired fractions are filtered to remove gels.
Claim 7
Dependent Claim
19 words
A method according to claim 1, wherein the process is repeated at least once more on the eluted product.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-04-01 | 1988-10-01 | 1989-04-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-04-01 | 1992-10-01 | 1993-04-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-04-01 | 1996-10-01 | 1997-04-02 | $7700.00 | Window Closed / Lapsed |