Recombinant expression of proteins from secretory cell lines
First Claim
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1. A method for the production of a polypeptide comprising the steps of:
- (a) providing a eukarvotic secretory host cell;
(b) inhibiting the production of an endogenous, secreted polypeptide;
(c) transforming said secretory host cell with an exogenous polynucleotide comprising a gene encoding an exogenous polypeptide, wherein said gene is under the control of a promoter active in eukaryotic cells; and
(d) culturing said secretory host cell under conditions such that said exogenous polynucleotide expresses said exogenous polypeptide;
wherein steps (a)-(c) are performed in vitro.
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Abstract
The present invention a provides methods for production of heterologous polypeptides using a variety recombinantly engineered secretory cell lines. The common feature of these cell lines is the absence of expression of at least one endogenous polypeptide. The host cell machinery normally used to produce the endogenous polypeptide is then usurped for the purpose of making the heterologous polypeptide. Also described are methods engineering cells for high level expression, methods of large scale protein production, and methods for treatment of disease in vivo using viral delivery systems and recombinant cell lines.
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Citations
26 Claims
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1. A method for the production of a polypeptide comprising the steps of:
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(a) providing a eukarvotic secretory host cell; (b) inhibiting the production of an endogenous, secreted polypeptide; (c) transforming said secretory host cell with an exogenous polynucleotide comprising a gene encoding an exogenous polypeptide, wherein said gene is under the control of a promoter active in eukaryotic cells; and (d) culturing said secretory host cell under conditions such that said exogenous polynucleotide expresses said exogenous polypeptide; wherein steps (a)-(c) are performed in vitro. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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Specification