Patent EP0117059A2 successfully refreshed from Google Patents.
EP0117059A2
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Methods for producing human tPA and expression vectors therefor
Assignee: Genentech Inc
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Filing: 1984-01-18
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Granted: —
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Expires: 2004-01-18
Abstract
Abstract A method and vectors for producing tissue plasminogen activator (tPA) in eukaryotic host cells is disclosed. Enhanced levels of tPA production are obtained by co-amplification of the tPA gene through treatment with methotrexate of cultures transformed with expression vectors coding additionally for mutant or wild type DHFR.
Inventors
- 👤 Arthur David Levinson
- 👤 Christian Clinton Simonsen
- 👤 Elizabeth Mcleod Yelverton
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 15:49
CPC Classifications
C; C12; C12N; C12N15/00; C12N9/00; C12N9/0004; C12N9/0012; C12N9/0026; C12N9/0028; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6456; C12N9/6459; C12Y; C12Y304/00; C12Y304/21; C12Y304/21069
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
41 words
An expression vector which comprises: a first DNA sequence encoding a DHFR protein; and a second DNA sequence encoding human tPA; wherein each of said first and second sequences is operably linked to a DNA sequence capable of effecting its expression.
Claim 2
Dependent Claim
41 words
An expression vector which comprises: a first DNA sequence encoding a DHFR protein; and a second DNA sequence encoding human tPA; wherein each of said first and second sequences is operably linked to a DNA sequence capable of effecting its expression.
Claim 3
Dependent Claim
33 words
A vector capable of effecting expression of a DNA sequence encoding human tPA and of a DNA sequence encoding a DHFR protein, said sequences being disposed therein, in cells transformed by said vector.
Claim 4
Dependent Claim
33 words
A vector capable of effecting expression of a DNA sequence encoding human tPA and of a DNA sequence encoding a DHFR protein, said sequences being disposed therein, in cells transformed by said vector.
Claim 5
Dependent Claim
13 words
The vector of Claim 1 wherein the DHFR protein is wild type DHFR.
Claim 6
Dependent Claim
13 words
The vector of Claim 3 which is effective in cells deficient in DHFR.
Claim 7
Dependent Claim
16 words
The vector of Claim 1 wherein the DHFR protein has a low binding affinity for MTX.
Claim 8
Dependent Claim
13 words
The vector of Claim 5 which is effective in cells sensitive to MTX.
Claim 9
Dependent Claim
5 words
The plasmid pETPER or pETPFR.
Claim 10
Dependent Claim
5 words
The plasmid pETPER or pETPFR.
Claim 11
Independent Claim
46 words
A method of preparing tPA in host cells which method comprises: transfecting said host cells with a vector effective in expressing DNA sequences for DHFR protein and for tPA disposed therein; growing the host cells under conditions suitable for said expression; recovering tPA from said cells.
Claim 12
Independent Claim
46 words
A method of preparing tPA in host cells which method comprises: transfecting said host cells with a vector effective in expressing DNA sequences for DHFR protein and for tPA disposed therein; growing the host cells under conditions suitable for said expression; recovering tPA from said cells.
Claim 13
Dependent Claim
22 words
A method according to claim 8 wherein the vector is as defined in any one of claims 1 and 3 to 7.
Claim 14
Dependent Claim
15 words
Cells transformed by the vector of any one of claims 1 and 3 to 7.
Claim 15
Independent Claim
44 words
A method of amplifying the DNA sequence encoding tPA in a host cell culture which method comprises providing the vector of any one of claims 1 and 3 to 7 and growing the cells in the presence of an effective amplifying concentration of MTX.
Claim 16
Independent Claim
43 words
A method of increasing the production of tPA in a host cell culture, which method comprises providing the vector of any one of claims 1 and 3 to 7 and growing the cells in the presence of an effective amplifying concentration of MTX.
References Cited (Backward Citations)
No citation records indexed.
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