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Abstract
Chinese hamster ovary (CHO) cells are the predominant production vehicle for biotherapeutics. Quantitative proteomics data were obtained from two CHO cell lines (CHO-S and CHO DG44) and compared with seven Chinese hamster (Cricetulus griseus) tissues (brain, heart, kidney, liver, lung, ovary and spleen) by tandem mass tag (TMT) labeling followed by mass spectrometry, providing a comprehensive hamster tissue and cell line proteomics atlas. Of the 8470 unique proteins identified, high similarity was observed between CHO-S and CHO DG44 and included increases in proteins involved in DNA replication, cell cycle, RNA processing, and chromosome processing. Alternatively, gene ontology and pathway analysis in tissues indicated increased protein intensities related to important tissue functionalities. Proteins enriched in the brain included those involved in acidic amino acid metabolism, Golgi apparatus, and ion and phospholipid transport. The lung showed enrichment in proteins involved in BCAA catabolism, ROS metabolism, vesicle trafficking, and lipid synthesis while the ovary exhibited enrichments in extracellular matrix and adhesion proteins. The heart proteome included vasoconstriction, complement activation, and lipoprotein metabolism enrichments. These detailed comparisons of CHO cell lines and hamster tissues will enhance understanding of the relationship between proteins and tissue function and pinpoint potential pathways of biotechnological relevance for future cell engineering.
Highlights
Chinese hamster ovary (CHO) cells are the predominant production vehicle for biotherapeutics
CHO-S cells were cultured in CD-CHO medium supplemented with 8 mM glutamine (ThermoFisher Scientific, Waltham, MA, USA), and CHO DG44 cells were cultured in DG44 medium supplemented with 2 mM glutamine (ThermoFisher Scientific, Waltham, MA, USA)
For both gene ontology (GO) and ingenuity pathway analysis (IPA), the enrichment and depletion values were determined based on p-values that were calculated via the hypergeometric distribution, with p < 0.05 set for evaluating significance
Summary
Chinese hamster ovary (CHO) cells are the predominant production vehicle for biotherapeutics. The heart proteome included vasoconstriction, complement activation, and lipoprotein metabolism enrichments. These detailed comparisons of CHO cell lines and hamster tissues will enhance understanding of the relationship between proteins and tissue function and pinpoint potential pathways of biotechnological relevance for future cell engineering. Initial efforts to understand CHO include the sequencing of both the CHO and Cricetulus griseus genomes. Initial CHO proteomic analyses revealed enrichment in protein processing and apoptosis pathways at the proteomic level in the CHO-K1 cell line compared to the human, mouse, and CHO genome and t ranscriptome[11]. Different bioprocess conditions have been studied via transcriptomics and proteomics to yield insights into protein production, cell growth, cell death, favorable glycosylation, and optimized media formulations[14,15]
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