Journal
ENGINEERING IN LIFE SCIENCES
Volume 17, Issue 7, Pages 801-808Publisher
WILEY
DOI: 10.1002/elsc.201600255
Keywords
alpha-1,6 fucosyltransferase (FUT8); Antibody-dependent cellular cytotoxicity (ADCC); CRISPR; Cas9; Defucosylated antibodies; Gene knockout
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Funding
- National Science Foundation of China [81502969]
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To engineer a host cell line that produces defucosylated mAbs with superior antibody dependent cellular cytotoxicity, we disrupted alpha-1,6 fucosyltransferase (FUT8) gene in CHO-S (CHO is Chinese hamster ovary) cells by clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9. The gene knockout cell line was evaluated for growth, stability, and product quality. The growth profile of FUT8 gene knockout CHO-S (FUT8(-/-)) cells was comparable with wild type CHO-S cells. FUT8 catalyzes the transfer of a fucose residue from GDP-fucose to N-glycans residue. Defucosylated IgG1 antibodies produced by FUT8(-/-) cells showed increased binding affinities to human Fc gamma RIIIa and higher activities in mediating antibody-dependent cellular cytotoxicity, comparing with conventional fucosylated IgG1. Our results demonstrated the potential of using the clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9 technology in cell line engineering for biopharmaceutical industrial applications.
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