期刊
BMC BIOTECHNOLOGY
卷 14, 期 -, 页码 -出版社
BIOMED CENTRAL LTD
DOI: 10.1186/1472-6750-14-15
关键词
CHO; mTOR; Monoclonal antibody; Phosphotidylinositol 3-kinase; Cell engineering
资金
- HEA, Ireland: PRTLI cycle 4- (Bio) Pharmaceutical and Pharmacological Sciences project
- Majlis Amanah Rakyat, MARA, Malaysia
Background: High recombinant protein productivity in mammalian cell lines is often associated with phenotypic changes in protein content, energy metabolism, and cell growth, but the key determinants that regulate productivity are still not clearly understood. The mammalian target of rapamycin (mTOR) signalling pathway has emerged as a central regulator for many cellular processes including cell growth, apoptosis, metabolism, and protein synthesis. This role of this pathway changes in response to diverse environmental cues and allows the upstream proteins that respond directly to extracellular signals (such as nutrient availability, energy status, and physical stresses) to communicate with downstream effectors which, in turn, regulate various essential cellular processes. Results: In this study, we have performed a transcriptomic analysis using a pathway-focused polymerase chain reaction (PCR) array to compare the expression of 84 target genes related to the mTOR signalling in two recombinant CHO cell lines with a 17.4-fold difference in specific monoclonal antibody productivity (q(p)). Eight differentially expressed genes that exhibited more than a 1.5-fold change were identified. Pik3cd (encoding the Class 1A catalytic subunit of phosphatidylinositol 3-kinase [PI3K]) was the most differentially expressed gene having a 71.3-fold higher level of expression in the high producer cell line than in the low producer. The difference in the gene's transcription levels was confirmed at the protein level by examining expression of p110 delta. Conclusion: Expression of p110 delta correlated with specific productivity (q(p)) across six different CHO cell lines, with a range of expression levels from 3 to 51 pg/cell/day, suggesting that p110 delta may be a key factor in regulating productivity in recombinant cell lines.
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