RRC ID 57750
著者 Fukuda N, Senga Y, Honda S.
タイトル Anxa2- and Ctsd-knockout CHO cell lines to diminish the risk of contamination with host cell proteins.
ジャーナル Biotechnol Prog
Abstract Chinese hamster ovary (CHO) cells have been used as host cells in the production of a range of recombinant therapeutic proteins, including monoclonal antibodies and Fc-fusion proteins. Host cell proteins (HCP) represent impurities that must be removed from therapeutic formulations because of their potential risks for immunogenicity. While the majority of HCP impurities are effectively removed in typical downstream purification processes, clearance of a small population of HCP remains challenging. In this study, we knocked out the Anxa2 and Ctsd genes to assess the feasibility of knockout approaches for diminishing the risk of contamination with HCP. Using the CRISPR/Cas9 system, Anxa2-, and Ctsd-knockout CHO cell lines were successfully established, and we confirmed the complete elimination of the corresponding HCP in cell lysates. Importantly, all knockout cell lines showed similar growth and viability to those of the wild-type control during 8 days of cultivation. Thus, knockout of unrequired genes can reduce contamination with HCP in the production of recombinant therapeutic proteins.
巻・号 35(4)
ページ e2820
公開日 2019-7-1
DOI 10.1002/btpr.2820
PMID 30972970
MeSH Animals Annexin A2 / chemistry* Annexin A2 / deficiency Annexin A2 / genetics Antibodies, Monoclonal / chemistry* CHO Cells Cathepsin D / chemistry* Cathepsin D / deficiency Cathepsin D / genetics Cell Survival Cells, Cultured Cricetulus Humans Recombinant Fusion Proteins / chemistry*
IF 2.334
引用数 3
リソース情報
ヒト・動物細胞 CHO-K1(RCB0285)