Patent US4784950A successfully refreshed from Google Patents.
US4784950A
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Expression of factor VII activity in mammalian cells
Assignee: Novo Nordisk Health Care AG
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Filing: 1986-12-16
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Granted: 1988-11-15
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Expires: 2006-12-16
Abstract
Abstract Methods are disclosed for producing proteins having biological activity for blood coagulation mediated by Factor VIIa. The proteins are produced by mammalian host cells which have been stably transfected with a DNA construct containing a nucleotide sequence which codes at least partially for either Factor VII. The nucleotide sequence comprises a first nucleotide sequence encoding a calcium binding domain, joined to a second nucleotide sequence positioned downstream of the first sequence. In particular, the first nucleotide sequence may be derived from a genomic clone or cDNA clone of Factor VII. The second sequence encodes a catalytic domain for the serine protease activity of Factor VIIA. The joined sequences code for proteins having substantially the same biological activity for blood coagulation as Factor VIIa.
Inventors
- 👤 Frederick S. Hagen
- 👤 Mark J. Murray
- 👤 Sharon J. Busby
- 👤 Kathleen L. Berkner
- 👤 Margaret Y. Insley
- 👤 Richard G. Woodbury
- 👤 Charles L. Gray
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 14:34
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6437; A; A61; A61P; A61P7/00; A61P7/04; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/755; C12N15/00; C12N15/09; C12N15/11; C12N15/62; C12N9/644; C12N9/647; C12Y; C12Y304/00; C12Y304/21; C12Y304/21021; C12Y304/21022; A61K; A61K38/00; C07K2319/00; C07K2319/20
Claims Analysis
Total Claims: 41
Claim 1
Independent Claim
10 words
A DNA construct comprising a DNA sequence encoding Factor VII.
Claim 2
Dependent Claim
10 words
A DNA construct comprising a DNA sequence encoding Factor VII.
Claim 3
Dependent Claim
24 words
The DNA construct of claim 1 wherein at least a portion of said DNA sequence is derived from a cDNA clone of Factor VII.
Claim 4
Dependent Claim
24 words
The DNA construct of claim 1 wherein at least a portion of said DNA sequence is derived from a genomic clone of Factor VII.
Claim 5
Dependent Claim
23 words
The DNA construct of claim 1 wherein said DNA sequence comprises the cDNA sequence of FIG. 1b, from bp 36 to bp 1433.
Claim 6
Dependent Claim
34 words
The DNA construct of claim 1 wherein said DNA sequence comprises the cDNA sequence of FIG. 1b, from bp 36 to bp 99, followed downstream by the sequence from bp 166 to bp 1433.
Claim 7
Dependent Claim
54 words
A DNA construct comprising a first nucleotide sequence joined to a second nucleotide sequence positioned downstream of said first sequence, said first and second sequences derived from cDNA clones of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa.
Claim 8
Dependent Claim
54 words
A DNA construct comprising a first nucleotide sequence joined to a second nucleotide sequence positioned downstream of said first sequence, said first and second sequences derived from cDNA clones of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa.
Claim 9
Dependent Claim
61 words
A DNA construct comprising a first nucleotide sequence derived from a genomic clone of Factor VII, joined to a second nucleotide sequence positioned downstream of said first sequence, said second sequence derived from a cDNA clone of Factor VII, the joined sequence coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa.
Claim 10
Dependent Claim
61 words
A DNA construct comprising a first nucleotide sequence derived from a genomic clone of Factor VII, joined to a second nucleotide sequence positioned downstream of said first sequence, said second sequence derived from a cDNA clone of Factor VII, the joined sequence coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa.
Claim 11
Dependent Claim
10 words
A recombinant plasmid comprising a DNA sequence encoding Factor VII.
Claim 12
Dependent Claim
10 words
A recombinant plasmid comprising a DNA sequence encoding Factor VII.
Claim 13
Dependent Claim
23 words
The recombinant plasmid of claim 8 wherein the DNA sequence comprises the cDNA sequence of FIG. 1b from bp 36 to bp 1433.
Claim 14
Dependent Claim
34 words
The recombinant plasmid of claim 8 wherein the DNA sequence comprises the cDNA sequence of FIG. 1b from bp 36 to bp 99, followed downstream by the sequence from bp 166 to bp 1433.
Claim 15
Dependent Claim
14 words
The recombinant plasmid of claim 8 wherein said DNA sequence comprises Factor VII cDNA.
Claim 16
Dependent Claim
15 words
The recombinant plasmid of claim 8 wherein said DNA sequence comprises Factor VII genomic DNA.
Claim 17
Dependent Claim
35 words
A recombinant plasmid capable of integration in mammalian host cell DNA, said plasmid comprising a promoter followed downstream by a DNA sequence encoding Factor VII, said DNA sequence being followed downstream by a polyadenylation signal.
Claim 18
Dependent Claim
35 words
A recombinant plasmid capable of integration in mammalian host cell DNA, said plasmid comprising a promoter followed downstream by a DNA sequence encoding Factor VII, said DNA sequence being followed downstream by a polyadenylation signal.
Claim 19
Dependent Claim
79 words
A recombinant plasmid capable of integration in mammalian host cell DNA, said plasmid comprising a promoter followed downstream by a first nucleotide sequence joined to a second nucleotide sequence positioned downstream of said first sequence, said first and second sequences derived from cDNA clones of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa, the joined sequences being followed downstream by a polyadenylation signal.
Claim 20
Dependent Claim
79 words
A recombinant plasmid capable of integration in mammalian host cell DNA, said plasmid comprising a promoter followed downstream by a first nucleotide sequence joined to a second nucleotide sequence positioned downstream of said first sequence, said first and second sequences derived from cDNA clones of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa, the joined sequences being followed downstream by a polyadenylation signal.
Claim 21
Dependent Claim
86 words
A recombinant plasmid capable of integration in mammalian host cell DNA, said plasmid comprising a promoter followed downstream by a first nucleotide sequence derived from a genomic clone of Factor VII, joined to a second nucleotide sequence positioned downstream of said first sequence, said second sequence derived from a cDNA clone of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa, the joined sequences being followed downstream by a polyadenylation signal.
Claim 22
Dependent Claim
86 words
A recombinant plasmid capable of integration in mammalian host cell DNA, said plasmid comprising a promoter followed downstream by a first nucleotide sequence derived from a genomic clone of Factor VII, joined to a second nucleotide sequence positioned downstream of said first sequence, said second sequence derived from a cDNA clone of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa, the joined sequences being followed downstream by a polyadenylation signal.
Claim 23
Dependent Claim
23 words
Mammalian cells stably transfected with a recombinant plasmid comprising a DNA sequence encoding Factor VII, said cells producing Factor VII in recoverable amounts.
Claim 24
Dependent Claim
23 words
Mammalian cells stably transfected with a recombinant plasmid comprising a DNA sequence encoding Factor VII, said cells producing Factor VII in recoverable amounts.
Claim 25
Dependent Claim
22 words
The cells of claim 16 wherein the DNA sequence comprises the cDNA sequence of FIG. 1b from bp 36 to bp 1433.
Claim 26
Dependent Claim
33 words
The cells of claim 16 wherein the DNA sequence comprises the cDNA sequence of FIG. 1b from bp 36 to bp 99, followed downstream by the sequence from bp 166 to bp 1433.
Claim 27
Dependent Claim
13 words
The cells of claim 16 wherein said DNA sequence comprises Factor VII cDNA.
Claim 28
Dependent Claim
14 words
The cells of claim 16 wherein said DNA sequence comprises Factor VII genomic DNA.
Claim 29
Dependent Claim
30 words
The cells of claim 16 wherein said plasmid comprises a promoter followed downstream by a DNA sequence encoding Factor VII, said DNA sequence being followed downstream by a polyadenylation signal.
Claim 30
Dependent Claim
74 words
The cells of claim 16 wherein said plasmid comprises a promoter followed downstream by a first nucleotide sequence joined to a second nucleotide sequence positioned downstream of said first sequence, said first and second sequences derived from cDNA clones of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa, the joined sequences being followed downstream by a polyadenylation signal.
Claim 31
Dependent Claim
81 words
The cells of claim 16 wherein said plasmid comprises a promoter followed downstream by a first nucleotide sequence derived from a genomic clone of Factor VII, joined to a second nucleotide sequence positioned downstream of said first sequence, said second sequence derived from a cDNA clone of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa, the joined sequences being followed downstream by a polyadenylation signal.
Claim 32
Independent Claim
67 words
A method for producing a protein having biological activity for blood coagulation mediated by Factor VIIa, comprising: establishing a mammalian host cell which contains a DNA construct comprising a DNA sequence encoding Factor VII; growing said mammalian host cell in an appropriate medium; isolating the protein product encoded by said DNA construct produced by said mammalian host cell; and activating said protein product to generate Factor VIIa.
Claim 33
Independent Claim
67 words
A method for producing a protein having biological activity for blood coagulation mediated by Factor VIIa, comprising: establishing a mammalian host cell which contains a DNA construct comprising a DNA sequence encoding Factor VII; growing said mammalian host cell in an appropriate medium; isolating the protein product encoded by said DNA construct produced by said mammalian host cell; and activating said protein product to generate Factor VIIa.
Claim 34
Dependent Claim
29 words
The method of claim 24, including amplification of the DNA sequence by cotransfection of the host cell with a gene encoding dihydrofolate reductase, wherein the appropriate medium comprises methotrexate.
Claim 35
Dependent Claim
34 words
The method of claim 24 wherein said protein product is activated by reacting the protein with a proteolytic enzyme selected from the group consisting of Factor XIIa, Factor IXa, kallikrein, Factor Xa, and thrombin.
Claim 36
Dependent Claim
13 words
The method of claim 24 wherein said DNA sequence comprises Factor VII cDNA.
Claim 37
Dependent Claim
14 words
The method of claim 24 wherein said DNA sequence comprises Factor VII genomic DNA.
Claim 38
Dependent Claim
22 words
The method of claim 24 wherein said DNA sequence comprises the cDNA sequence of FIG. 1b, from bp 36 to bp 1433.
Claim 39
Dependent Claim
33 words
The method of claim 24 wherein said DNA sequence comprises the cDNA sequence of FIG. 1b, from bp 36 to bp 99, followed downstream by the sequence from bp 166 to bp 1433.
Claim 40
Dependent Claim
60 words
The method of claim 24 wherein said DNA sequence comprises a first nucleotide sequence joined to a second nucleotide sequence positioned downstream of said first sequence, said first and second sequences derived from cDNA clones of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa.
Claim 41
Dependent Claim
67 words
The method of claim 24 wherein said DNA sequence comprises a first nucleotide sequence derived from a genomic clone of Factor VII, joined to a second nucleotide sequence positioned downstream of said first sequence, said second sequence derived from a cDNA clone of Factor VII, the joined sequences coding for a protein which upon activation has substantially the same biological activity for blood coagulation as Factor VIIa.
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) | 1991-11-15 | 1992-05-16 | 1992-11-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-11-15 | 1996-05-16 | 1996-11-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-11-15 | 2000-05-16 | 2000-11-15 | $7700.00 | Window Closed / Lapsed |