US5364771A
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Hybrid human/porcine factor VIII
Assignee: Emory University
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Filing: 1992-04-07
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Granted: 1994-11-15
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Expires: 2012-04-07
Abstract
Abstract A hybrid human/porcine coagulation factor VIII is produced by isolation and recombination of human and porcine factor VIII subunits, or by genetic engineering of the human and porcine factor VIII genes. Subunits of factor VIII that have been purified from human or porcine plasma are isolated, and hybrid human/porcine factor VIII is produced by mixing either porcine heavy chain subunits with human light chain subunits or by mixing human heavy chain subunits with porcine light chain subunits, thereby producing human light chain/porcine heavy chain and human heavy chain/porcine light chain hybrid molecules. These hybrid molecules are isolated by ion exchange chromatography. Alternatively, recombinant DNA methods are used to swap elements of porcine factor VIII for the corresponding elements of human factor VIII to produce hybrid human/porcine factor VIII.
Inventors
- 👤 John S. Lollar
- 👤 Marschall S. Runge
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-22 23:52
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/755; A; A61; A61P; A61P7/00; A61P7/04; A61K; A61K38/00; C07K2319/00; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/10
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
127 words
A purified hybrid factor VIII molecule comprising porcine and human amino acid sequences, wherein the molecule has procoagulant activity in an in vitro coagulation assay and wherein the molecule is selected from the group consisting of a molecule consisting essentially of a human factor VIII in which the porcine A2 domain is substituted for the homologous human factor VIII A2 domain; a molecule consisting essentially of a porcine factor VIII in which the human A2 domain is substituted for the homologous porcine factor VIII A2 domain; a molecule consisting essentially of a human light chain factor VIII subunit and a porcine heavy chain factor VIII subunit; and a molecule consisting essentially of a porcine light chain factor VIII subunit and a human heavy chain factor VIII subunit.
Claim 2
Dependent Claim
127 words
A purified hybrid factor VIII molecule comprising porcine and human amino acid sequences, wherein the molecule has procoagulant activity in an in vitro coagulation assay and wherein the molecule is selected from the group consisting of a molecule consisting essentially of a human factor VIII in which the porcine A2 domain is substituted for the homologous human factor VIII A2 domain; a molecule consisting essentially of a porcine factor VIII in which the human A2 domain is substituted for the homologous porcine factor VIII A2 domain; a molecule consisting essentially of a human light chain factor VIII subunit and a porcine heavy chain factor VIII subunit; and a molecule consisting essentially of a porcine light chain factor VIII subunit and a human heavy chain factor VIII subunit.
Claim 3
Dependent Claim
34 words
The molecule of claim 1, wherein the molecule has a specific activity greater than 20,000 U/A 280 protein in aqueous solution when human plasma is used as the standard in a one-stage coagulation assay.
Claim 4
Dependent Claim
18 words
The molecule of claim 1, wherein the hybrid human/procine factor VIII is combined with a pharmaceutically acceptable carrier.
Claim 5
Dependent Claim
20 words
The molecular of claim 3, wherein the carrier is selected from the group consisting of stabilizing agents and delivery vehicles.
Claim 6
Dependent Claim
19 words
The molecule of claim 4, wherein the stabilizing agents are selected from the group consisting of proteins and polysaccharides.
Claim 7
Dependent Claim
27 words
The molecule of claim 3, further comprising clotting factors selected from the group consisting of von Willebrand factor, vitamin K dependent clotting factors, and coagulant tissue factor.
Claim 8
Dependent Claim
11 words
The molecule of claim 4, wherein the delivery vehicles are liposomes.
Claim 9
Independent Claim
97 words
A method of preparing puirified hybrid human/porcine factor VIII comprising combining primary amino acid sequence derived from procine factor VIII with primary amino acid sequence derived from human factor VIII to form a hybrid factor VIII molecule having procoagulant activity in an in vitro coagulation assay, wherein the molecule is selected from the group consisting of a molecule consisting essentially of a human light chain factor VIII subunit and a procine heavy chain factor VIII subunit; and a molecule consisting essentially of a porcine light chain factor VIII subunit and a human heavy chain factor VIII subunit.
Claim 10
Independent Claim
97 words
A method of preparing puirified hybrid human/porcine factor VIII comprising combining primary amino acid sequence derived from procine factor VIII with primary amino acid sequence derived from human factor VIII to form a hybrid factor VIII molecule having procoagulant activity in an in vitro coagulation assay, wherein the molecule is selected from the group consisting of a molecule consisting essentially of a human light chain factor VIII subunit and a procine heavy chain factor VIII subunit; and a molecule consisting essentially of a porcine light chain factor VIII subunit and a human heavy chain factor VIII subunit.
Claim 11
Dependent Claim
45 words
The method of claim 8, wherein the hybrid human/porcine factor VIII molecule is formed by isolating and purifying heavy and light chain subunits of human factor VIII and porcine factor VIII, then mixing the human and porcine subunits to form the hybrid human/porcine factor VIII.
Claim 12
Dependent Claim
20 words
The method of claim 9, wherein the human and porcine factor VIII subunits are isolated from human and porcine plasma.
Claim 13
Independent Claim
78 words
A method of preparing purified hybrid human/porcine factor VIII comprising expressing recombinant DNA encoding domains in the light chain and heavy chain subunits of porcine and human factor VIII, further comprising substituting A2 domains of porcine and human factor VIII, to form the purified hybrid human/porcine factor VIII having primary amino acid sequence derived from porcine factor VIII and primary amino acid sequence derived from human factor VIII and having procoagulant activity in an in vitro coagulation assay.
Claim 14
Independent Claim
78 words
A method of preparing purified hybrid human/porcine factor VIII comprising expressing recombinant DNA encoding domains in the light chain and heavy chain subunits of porcine and human factor VIII, further comprising substituting A2 domains of porcine and human factor VIII, to form the purified hybrid human/porcine factor VIII having primary amino acid sequence derived from porcine factor VIII and primary amino acid sequence derived from human factor VIII and having procoagulant activity in an in vitro coagulation assay.
Claim 15
Dependent Claim
29 words
The method of claim 9, wherein the hybrid human/porcine factor VIII molecule is formed by mixing human light chain factor VIII subunits and porcine heavy chain factor VIII subunits.
Claim 16
Dependent Claim
29 words
The method of claim 9, wherein the hybrid human/porcine factor VIII molecule is formed by mixing porcine light chain factor VIII subunits and human heavy chain factor VIII subunits.
Claim 17
Dependent Claim
31 words
The molecule of claim 1, wherein the molecule consists essentially of a human factor VIII in which the porcine A2 domain is substituted for the homologous human factor VIII A2 domain.
Claim 18
Dependent Claim
31 words
The molecule of claim 1, wherein the molecule consists essentially of a porcine factor VIII in which the human A2 domain is substituted for the homologous porcine factor VIII A2 domain.
Claim 19
Dependent Claim
26 words
The molecule of claim 1, wherein the molecule consists essentially of a human light chain factor VIII subunit and a porcine heavy chain factor VIII subunit.
Claim 20
Dependent Claim
26 words
The molecule of claim 1, wherein the molecule consists essentially of a porcine light chain factor VIII subunit and a human heavy chain factor VIII subunit.
Claim 21
Dependent Claim
24 words
The method of claim 11, wherein the hybrid human/porcine factor VIII molecule is formed by substituting the porcine A2 domain in human factor VIII.
Claim 22
Dependent Claim
24 words
The method of claim 11, wherein the hybrid human/porcine factor VIII molecule is formed by substituting the human A2 domain in porcine factor VIII.
References Cited (Backward Citations)
USRE32011E
Cited
US4757006A
Cited
US4965199A
Cited
US4970300A
Cited
US4980456A
Cited
US5004803A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1997-11-14 | 1998-05-16 | 1998-11-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-11-14 | 2002-05-16 | 2002-11-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-11-14 | 2006-05-16 | 2006-11-15 | $7700.00 | Window Closed / Lapsed |