| Literature DB >> 28544881 |
Faraaz Noor Khan Yusufi1, Meiyappan Lakshmanan1, Ying Swan Ho1, Bernard Liat Wen Loo1, Pramila Ariyaratne2, Yuansheng Yang1, Say Kong Ng1, Tessa Rui Min Tan1, Hock Chuan Yeo3, Hsueh Lee Lim1, Sze Wai Ng1, Ai Ping Hiu1, Chung Ping Chow1, Corrine Wan1, Shuwen Chen1, Gavin Teo1, Gao Song2, Ju Xin Chin1, Xiaoan Ruan2, Ken Wing Kin Sung2, Wei-Shou Hu4, Miranda Gek Sim Yap3, Muriel Bardor5, Niranjan Nagarajan6, Dong-Yup Lee7.
Abstract
Effective development of host cells for therapeutic protein production is hampered by the poor characterization of cellular transfection. Here, we employed a multi-omics-based systems biotechnology approach to elucidate the genotypic and phenotypic differences between a wild-type and recombinant antibody-producing Chinese hamster ovary (CHO) cell line. At the genomic level, we observed extensive rearrangements in specific targeted loci linked to transgene integration sites. Transcriptional re-wiring of DNA damage repair and cellular metabolism in the antibody producer, via changes in gene copy numbers, was also detected. Subsequent integration of transcriptomic data with a genome-scale metabolic model showed a substantial increase in energy metabolism in the antibody producer. Metabolomics, lipidomics, and glycomics analyses revealed an elevation in long-chain lipid species, potentially associated with protein transport and secretion requirements, and a surprising stability of N-glycosylation profiles between both cell lines. Overall, the proposed knowledge-based systems biotechnology framework can further accelerate mammalian cell-line engineering in a targeted manner.Entities:
Keywords: Chinese hamster ovary; genome-scale metabolic model; genomic rearrangements; glycosylation; lipid metabolism; monoclonal antibody-producing cell line; systems biotechnology; tricistronic vector
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Year: 2017 PMID： 28544881 DOI： 10.1016/j.cels.2017.04.009
Source DB: PubMed Journal: Cell Syst ISSN： 2405-4712 Impact factor: 10.304