Patent US4229540A successfully refreshed from Google Patents.
US4229540A
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Hydrolase purified from human plasma
Assignee: Bayer Corp
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Filing: 1979-07-02
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Granted: 1980-10-21
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Expires: 1999-07-02
Abstract
Abstract A previously undescribed hydrolytic enzyme has been successfully isolated from human plasma and purified to homogeneity. The enzyme has a molecular weight of about 70,000 daltons, has a Km for the substrate H-D-Phe-Pip-Arg-PNA of about 0.057 mM, a KI for benzamidine of about 1.1x10-5 M, is bindable to and elutable from insoluble barium salts, is inhibited by antithrombin III, especially in the presence of heparin, and, in vitro, has been found to activate and degrade human Factor VIII, activate human Factor V, and inhibit platelet aggregation induced by ADP, epinephrine, or collagen, thus providing a substance useful for antithrombotic studies.
Inventors
- 👤 Michael H. Coan
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 17:48
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/56; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; A; A61; A61K; A61K38/00
Claims Analysis
Total Claims: 4
Claim 1
Independent Claim
88 words
An enzyme preparation comprising a substantially homogeneous hydrolytic enzyme characterized by being obtainable from human plasma, having a molecular weight of about 70,000 daltons as determined by comparative polyacrylamide gel electrophoresis, having a K m for the substrate H-D-Phe-Pip-Arg-PNA of about 0.057 mM, having a K I for benzamidine of about 1.1×10 -5 M, being bindable to and elutable from insoluble barium salts, and, in vitro, capable of activating and degrading human Factor VIII, activating human Factor V, and inhibiting platelet aggregation induced by epinephrine, ADP, or collagen.
Claim 2
Dependent Claim
88 words
An enzyme preparation comprising a substantially homogeneous hydrolytic enzyme characterized by being obtainable from human plasma, having a molecular weight of about 70,000 daltons as determined by comparative polyacrylamide gel electrophoresis, having a K m for the substrate H-D-Phe-Pip-Arg-PNA of about 0.057 mM, having a K I for benzamidine of about 1.1×10 -5 M, being bindable to and elutable from insoluble barium salts, and, in vitro, capable of activating and degrading human Factor VIII, activating human Factor V, and inhibiting platelet aggregation induced by epinephrine, ADP, or collagen.
Claim 3
Dependent Claim
25 words
The enzyme preparation of claim 1 wherein the enzyme, upon reduction, yields two peptide chains having respective molecular weights of about 25,000 and 45,000 daltons.
Claim 4
Dependent Claim
73 words
A method of isolating a hydrolase enzyme as defined in claim 1 from human plasma which comprises the steps of a. passing a human Factor IX concentrate through an immobilized benzamidine affinity chromatography column; b. eluting the product held thereby and collecting it to form a first isolation product; c. passing the first isolation product through an immobilized heparin affinity chromatography column; and d. eluting the product held thereby to isolate the enzyme.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-10-21 | 1984-04-21 | 1984-10-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-10-21 | 1988-04-21 | 1988-10-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-10-21 | 1992-04-21 | 1992-10-21 | $7700.00 | Window Closed / Lapsed |