title: Proteomic studies support the use of multi-product immunoassays to monitor host cell protein impurities.
abstract: In the biopharmaceutical industry, recombinant protein drugs are commonly produced in Chinese hamster ovary (CHO) cells. During the development process, removal of CHO cell-derived proteins from the biopharmaceutical product is monitored using multi-product immunoassays. Such immunoassays are developed by raising antibodies to a single CHO cell protein preparation. However, these assays are utilized to monitor CHO cell protein impurities during the recovery of products from different CHO cell lines. To address whether underlying differences between CHO cell lines result in sufficient protein expression changes to exclude the suitability of multi-product immunoassays, a comparative proteomics study of three independently generated CHO cell lines was performed. Statistical analysis of over 1000 proteins resolved by 2-D PAGE demonstrated that the protein expression profiles of three different CHO cell lines exhibit very few differences in protein expression. Only 11 qualitative changes in protein expression and 26 quantitative changes greater than two-fold were observed. Identification of protein spots by mass spectrometry revealed that many of the observed changes were due to post-translational modifications rather than expression of novel proteins in each cell line. These results suggest that multi-product immunoassays are suitable for monitoring host cell proteins in biopharmaceuticals produced in different CHO cell lines.
authors.affiliation: Department of Early-Stage Analytical Development, Genentech Inc., MS #62, 1 DNA Way, South San Francisco, CA 94080, USA. denisek@gene.com
authors.forename: Denise C
authors.initials: DC
authors.lastname: Krawitz
authors.forename: William
authors.initials: W
authors.lastname: Forrest
authors.forename: G Tony
authors.initials: GT
authors.lastname: Moreno
authors.forename: Joshua
authors.initials: J
authors.lastname: Kittleson
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Champion
mesh_terms: D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002455:Cell Division; D002469:Cell Separation; D006224:Cricetinae; D017931:DNA Primers; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D007118:Immunoassay / Q000379:methods*; D020543:Proteome; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics
chemical_list: D017931:DNA Primers; D020543:Proteome; D013762:Tetrahydrofolate Dehydrogenase