US5527692A
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Methods for producing thrombin
Assignee: International Business Machines Corp
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Filing: 1995-06-05
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Granted: 1996-06-18
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Expires: 2015-06-05
Abstract
Abstract Methods are disclosed for producing thrombin. The protein is produced from host cells transformed or transfected with DNA construct(s) containing information necessary to direct the expression of thrombin precursors. The DNA constructs generally include the following operably linked elements: a transcriptional promoter, DNA sequence encoding a gla-domainless prothrombin, and a transcriptional terminator. Thrombin precursors produced from transformed or transfected host cells are activated either in vivo or in vitro.
Inventors
- 👤 Richard D. Holly
- 👤 Donald C. Foster
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 11:48
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6429; C07; C07K; C07K14/00; C07K14/37; C07K14/39; C12N9/6456; C12Y; C12Y304/00; C12Y304/21; C12Y304/21005; A; A61; A61K; A61K38/00; C07K2319/00; C07K2319/01; C07K2319/02
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
15 words
A pharmaceutical composition which comprises an activated recombinant gla-domainless prothrombin and a physiologically acceptable carrier.
Claim 2
Dependent Claim
15 words
A pharmaceutical composition which comprises an activated recombinant gla-domainless prothrombin and a physiologically acceptable carrier.
Claim 3
Dependent Claim
44 words
The pharmaceutical composition of claim 1, wherein the activated recombinant prothrombin is produced by cleavage of a thrombin precursor encoded by a DNA construct comprising the following operably linked elements: a transcriptional promoter, a DNA segment encoding a gla-domainless prothrombin and a transcriptional terminator.
Claim 4
Dependent Claim
39 words
The pharmaceutical composition of claim 2, wherein the promoter of the DNA construct encoding the thrombin precursor is the mouse metallothionein-1 promoter, the adenovirus major late promoter, the Saccharomyces cerevisiae TPI1 promoter or the Saccharomyces cerevisiae ADH2-4 c promoter.
Claim 5
Dependent Claim
48 words
The pharmaceutical composition of claim 2, wherein the DNA construct encoding the thrombin precursor further comprises a signal sequence selected from human tissue plasminogen activator leader sequence, the human α-2 plasmin inhibitor signal sequence, the Saccharomyces cerevisiae BAR1 secretory signal sequence or the Saccharomyces cerevisiae MFα1 signal sequence.
Claim 6
Dependent Claim
57 words
The pharmaceutical composition of claim 2, wherein the gla-domainless prothrombin comprises kringle 1, kringle 2, the A chain, an activation site and the serine protease domain of prothrombin; kringle 2, the A chain, an activation site and the serine protease domain of prothrombin; or the A chain, an activation site and the serine protease domain of prothrombin.
Claim 7
Dependent Claim
27 words
The pharmaceutical composition of claim 5, wherein the gla-domainless prothrombin activation site is cleavable by a snake venom activator, thrombin or the Saccharomyces cerevisiae KEX2 gene product.
Claim 8
Dependent Claim
15 words
The pharmaceutical composition of claim 1, wherein the recombinant gla-domainless prothrombin is human gla-domainless prothrombin.
Claim 9
Dependent Claim
15 words
The pharmaceutical composition of claim 2, wherein the recombinant thrombin precursor is human gla-domainless prothrombin.
Claim 10
Dependent Claim
9 words
The pharmaceutical composition of claim 1, further comprising fibrinogen.
Claim 11
Dependent Claim
19 words
An isolated protein which comprises recombinant human gla-domainless prothrombin that can be cleaved to form an active thrombin molecule.
Claim 12
Dependent Claim
19 words
An isolated protein which comprises recombinant human gla-domainless prothrombin that can be cleaved to form an active thrombin molecule.
Claim 13
Dependent Claim
24 words
A pharmaceutical composition which comprises recombinant human gla-domainless prothrombin that can be cleaved to form an active thrombin molecule, and a physiologically acceptable carrier.
Claim 14
Dependent Claim
24 words
A pharmaceutical composition which comprises recombinant human gla-domainless prothrombin that can be cleaved to form an active thrombin molecule, and a physiologically acceptable carrier.
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) | 1999-06-18 | 1999-12-18 | 2000-06-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-06-18 | 2003-12-18 | 2004-06-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-06-18 | 2007-12-18 | 2008-06-18 | $7700.00 | Window Closed / Lapsed |