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HIF‐1 Signaling Pathway Implicated in Phenotypic Instability in a Chinese Hamster Ovary Production Cell Line
Author(s) -
Lieske Paulena L.,
Wei Wei,
Crowe Kerstin B.,
Figueroa Bruno,
Zhang Lin
Publication year - 2020
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201900306
Subject(s) - chinese hamster ovary cell , cell culture , biology , microbiology and biotechnology , phenotype , cell , cell growth , genetics , gene
Monitoring genotypic and phenotypic stability is crucial during the development of recombinant Chinese hamster ovary (CHO) cell lines. Although genotypic instability is well‐studied, there are few reports on phenotypic instability. Here, a case study of two clonal cell lines derived from Pfizer's site‐specific integration expression platform that expresses the same monoclonal antibody is described. It is shown that both cell lines (herein referred to as “Cell Line A” and “Cell Line B”) are genotypically stable up to 130 generations. However, when both cell lines are run side‐by‐side in a fed‐batch production assay, productivity from Cell Line A later generation cells is much lower when compared to earlier generation cells. Phenotypically, later generation Cell Line A cells display increased lactate production, decreased productivity, and decreased cell viability. Metabolic analysis reveals that Cell Line A exhibits increased glycolysis activity and capacity at higher generational age. Whole transcriptomic sequencing shows significant upregulation of the hypoxia‐inducible factor 1‐alpha (HIF‐1α) signaling pathway and associated downstream targets. Furthermore, Western blot analysis confirms elevated HIF‐1α protein in Cell Line A cells at later generation. These results suggest a novel role for HIF‐1α in the age‐associated metabolic changes that result in the phenotypic instability of a recombinant CHO cell line.