EP0319944A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Co-expression in eukaryotic cells
Assignee: Zymogenetics Inc
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Filing: 1988-12-07
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Granted: —
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Expires: 2008-12-07
Abstract
Abstract A method of producing proteins of interest is disclosed. The method includes the introduction of a first DNA sequence encoding the protein of interest and at least one additional DNA sequence encoding a protein which processes or stabilizes the protein of interest into a eukaryotic host cell. The host cell is subsequently cultured under conditions which allow the DNA sequences to be expressed. Suitable eukaryotic hosts include mammalian cells and yeast cells.
Inventors
- 👤 Eileen R. Mulvihill
- 👤 Kathleen L. Berkner
- 👤 Donald C. Foster
- 👤 Ashok A. Kumar
- 👤 Vivian L. Mackay
- 👤 Gary E. Parker
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 22:11
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/647; C07; C07K; C07K14/00; C07K14/81; C07K14/8107; C07K14/811; C07K14/8121; C07K14/8125; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/79; C12N15/80; C12N15/81; C12N15/815; C12N15/85; C12N9/6435; C12N9/6456; C12N9/6459; C12Y; C12Y304/00; C12Y304/21; C12Y304/21007; C12Y304/21069
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
39 words
A eukaryotic host cell transfected or transformed with a first DNA sequence encoding a protein of interest and at least one additional DNA sequence, said additional DNA sequence encoding a protein which processes or stabilizes the protein of interest.
Claim 2
Dependent Claim
39 words
A eukaryotic host cell transfected or transformed with a first DNA sequence encoding a protein of interest and at least one additional DNA sequence, said additional DNA sequence encoding a protein which processes or stabilizes the protein of interest.
Claim 3
Dependent Claim
21 words
The host cell of claim 1 wherein the eukaryotic host cell is a mammalian host cell or a yeast host cell.
Claim 4
Dependent Claim
41 words
The host cell of claim 1 wherein the first DNA sequence is selected from the group consisting of sequences encoding t-PA, factor VII, factor IX, factor X, protein C, activated protein C, protein S, plasminogen, insulin and derivatives and analogs thereof.
Claim 5
Dependent Claim
15 words
The host cell of claim 1 wherein the first DNA sequence encodes a serine protease.
Claim 6
Dependent Claim
32 words
The host cell of claim 1 wherein the additional DNA sequence is selected from the group consisting of sequences encoding proteases, protease inhibitors, and proteins which bind to the protein of interest.
Claim 7
Dependent Claim
22 words
The host cell of claim 1 wherein the first DNA sequence encodes factor VII and the additional DNA sequence encodes factor IX.
Claim 8
Dependent Claim
28 words
The host cell of claim 1 wherein the first DNA sequence encodes protein C or activated protein C and the additional DNA sequence is the yeast KEX2 gene.
Claim 9
Dependent Claim
33 words
The host cell of claim 1 wherein the first DNA sequence encodes t-PA and the additional DNA sequence encodes a protease inhibitor selected from the group consisting of TIMP, trypsin inhibitors and aprotinin.
Claim 10
Dependent Claim
24 words
The host cell of claim 1 wherein the first DNA sequence encodes plasminogen and the additional DNA sequence encodes alpha-1-antitrypsin or a variant thereof.
Claim 11
Dependent Claim
22 words
The host cell of claim 1 wherein the first DNA sequence encodes protein C and the additional DNA sequence encodes protein S.
Claim 12
Dependent Claim
22 words
The host cell of claim 1 wherein the first DNA sequence encodes factor VII and the additional DNA sequence encodes tissue factor.
Claim 13
Independent Claim
45 words
A method for producing a protein of interest, comprising: culturing a host cell according to any of claims 1-11 under conditions which allow the first DNA sequence and the additional DNA sequence(s) to be expressed; and isolating the protein of interest from the host cell.
Claim 14
Dependent Claim
27 words
The method of claim 12 including, prior to the step of culturing, cotransfecting or cotransforming the host cell with multiple vectors, each containing a separate expression unit.
Claim 15
Dependent Claim
26 words
The method of claim 12 including, prior to the step of culturing, transfecting or transforming the host cell with a single vector containing multiple expression units.
Claim 16
Dependent Claim
40 words
The method of claim 12 including, prior to the step of culturing, transfecting the host cell with a single vector containing a single expression unit that is transcribed into a polycistronic message, wherein the host cell is a mammalian cell.
Claim 17
Dependent Claim
69 words
A DNA sequence which codes for a protein having substantially the same biological activity as human activated protein C, said sequence further coding for the amino acid sequence L-R₁-R₂-R₃-R₄-X-R₅-R₆-R₇-R₈-H, wherein L is essentially the light chain of activated protein C, R₁-R₈ are Lys or Arg, X is a peptide bond or a spacer peptide of 1-12 amino acids, and H is essentially the heavy chain of activated protein C.
Claim 18
Dependent Claim
69 words
A DNA sequence which codes for a protein having substantially the same biological activity as human activated protein C, said sequence further coding for the amino acid sequence L-R₁-R₂-R₃-R₄-X-R₅-R₆-R₇-R₈-H, wherein L is essentially the light chain of activated protein C, R₁-R₈ are Lys or Arg, X is a peptide bond or a spacer peptide of 1-12 amino acids, and H is essentially the heavy chain of activated protein C.
Claim 19
Dependent Claim
31 words
Mammalian cells transfected with an expression vector capable of integration in mammalian host cell DNA, said expression vector comprising a promoter operably linked to a DNA sequence according to claim 16.
Claim 20
Dependent Claim
22 words
The mammalian cells of claim 17 wherein said cells are further transfected with the KEX1 or KEX2 gene of Saccharomyces cerevisiae .
References Cited (Backward Citations)
No citation records indexed.
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