US4880915A
Expired / Lapsed
US
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Method for purifying a physiologically active substance produced by recombinant DNA technique
Assignee: Asahi Kasei Corp
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Filing: 1985-11-19
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Granted: 1989-11-14
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Expires: 2005-11-19
Abstract
Abstract An aqueous solution containing a physiologically active substance, which is produced by recombinant DNA technique and which has cytotoxic activity against L-M cells and is capable of inducing hemorrhagic necrosis of transplanted Meth A sarcoma in the BALB/c mouse, can be effectively, efficiently purified by column chromatography using a column packed with a dye-bonded crosslinked agarose gel.
Inventors
- 👤 Junichi Kajihara
- 👤 Takao Kiyota
- 👤 Hiroshi Hayashi
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 22:17
CPC Classifications
C; C12; C12N; C12N15/00; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/525; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/828
Claims Analysis
Total Claims: 5
Claim 1
Independent Claim
105 words
A method for purifying a human TNF which comprises: applying an aqueous solution containing human TNF to a column packed with a dye-bonded crosslinked agarose gel comprised of a crosslinked agarose gel support and a dye ligand covalently bonded thereto, said dye being a member selected from the group consisting of Cibacron Blue F3GA (color index name: reactive blue 2), Procion Red HE3B (color index name: reactive red 120), and Green A, whereby said human TNF is retained by said gel; eluting said column with an eluent to release said human TNF from said gel; and recovering a column fraction comprised of purified human TNF.
Claim 2
Independent Claim
105 words
A method for purifying a human TNF which comprises: applying an aqueous solution containing human TNF to a column packed with a dye-bonded crosslinked agarose gel comprised of a crosslinked agarose gel support and a dye ligand covalently bonded thereto, said dye being a member selected from the group consisting of Cibacron Blue F3GA (color index name: reactive blue 2), Procion Red HE3B (color index name: reactive red 120), and Green A, whereby said human TNF is retained by said gel; eluting said column with an eluent to release said human TNF from said gel; and recovering a column fraction comprised of purified human TNF.
Claim 3
Dependent Claim
41 words
The method according to claim 1, wherein said column packed with a dye-bonded crosslinked agarose gel is equilibrated with 0.1MNaCl buffer prior to applying said aqueous solution containing human TNF and said eluent is a buffer containing 0.5M or more NaCl.
Claim 4
Dependent Claim
45 words
The method according to claim 1, wherein said packed column is equilibrated to a pH of 5.5 to 6.5, said aqueous solution containing human TNF has a pH of 5.5 to 6.5, and said eluent comprises a buffer having a pH of 8.0 or greater.
Claim 5
Dependent Claim
15 words
The method according to claim 1, wherein said human TNF is a recombinant human TNF.
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) | 1992-11-13 | 1993-05-15 | 1993-11-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-11-13 | 1997-05-15 | 1997-11-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-11-13 | 2001-05-15 | 2001-11-14 | $7700.00 | Window Closed / Lapsed |