Homogeneity and secretion of recombinant proteins in mammalian systems
AssigneeApplied Research Systems ARS Holding N.V.
InventorsEdith Chitlaru, Hagit Amitai, Daniel Helman
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Abstract
The present invention relates to a method for improving homogeneity and/or secretion of a recombinant protein of interest expressed in mammalian cells by replacing the endogenous signal peptide sequence of the DNA encoding the protein of interest with that of human hGH. Specifically, the present invention relates to a method wherein the protein of interest is a subunit of the follicle stimulating hormone (FSH). The invention also relates to DNA expression vectors containing the sequence encoding such proteins of interest fused to the signal peptide sequence of the hGH and to cells harbouring such vectors.
Claims28
A method for improving the homogeneity of production of a recombinant protein of interest composed of more than one subunit in a mammalian cell, comprising the step of substituting the endogenous signal peptide of at least one of the subunits with an amino acid sequence comprising the signal peptide of human growth hormone (hGH).
The method according to anyone of the preceding claims, wherein the protein of interest is selected from thyrotropin (TSH), follicle stimulating hormone (FSH), luteinizing hormone (LH) and human choriogonadotropin (HCG).
The method according to anyone of the preceding claims, wherein the mammalian cell is selected from 3T3 cells, COS cells, human osteosarcoma cells, MRC-5 cells, BHK cells, VERO cells, CHO cells, rCHO-tPA cells, rCHO-Hep B Surface Antigen cells, HEK 293 cells, rHEK 293 cells, rC127-Hep B Surface Antigen cells, Normal Human fibroblast cells, Stroma cells, Hepatocytes cells, PER.C6 cells and human permanent amniocytic cells.
An expression vector for improving homogeneity of production of a recombinant protein of interest composed of more than one subunit, comprising the DNA sequence of one or more subunit, wherein the DNA sequence of at least one subunit's endogenous signal peptide is replaced with the DNA encoding the sequence of the hGH signal peptide.
Dependencies are inferred from the claim text.