Complement component 1s (C1s) deficient cells for production of vaccines and biopharmaceutical proteins
AssigneeThe Regents of the University of California
InventorsSophia Li, Phillip Berman
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Abstract
The present disclosure reports that a calcium-dependent serine protease, complement component is (C1s) has been identified as a protease responsive for cleavage of exogenous polypeptides expressed in mammalian cell lines such as CHO cells. These CHO cell lines provide for increased yield of antigenically correct, uncleaved exogenous polypeptide as compared to unmodified CHO cells expressing the active C1s protease. C1s-deficient cell lines and methods for use of same for producing exogenous polypeptides, e.g., human immunodeficiency virus (HIV) envelope glycoprotein polypeptides, such as, gp120 or human Factor VIII are provided.
Claims19
A genetically modified mammalian cell line comprising:
- a deletion in an endogenous c1s gene encoding a calcium-dependent serine (C1s) protease, wherein the deletion decreases calcium-dependent serine protease activity of the C1s protease, wherein the deletion consists of a deletion of (i) nucleotides 1448-1451 or (ii) nucleotides at positions 1444-1445 and 1447 with reference to wild type c1s mRNA sequence set forth in SEQ ID NO:4;
- and a heterologous nucleic acid comprising a nucleotide sequence encoding a gp120 polypeptide, wherein the gp120 polypeptide comprises a cleavage site for the C1s protease.
Dependencies are inferred from the claim text.