BIOTECHNOLOGY
Fig. 2. Fucosyltransferase-8 (FUT8) catalyses the transfer of fucose from GDP-fucose to N-linked type complex glycopeptides
techniques, it is now possible to create endogenous FUT8 knockout cell lines to generate fucose-free antibodies. Humanised IgG1 antibodies expressed in fucose-free CHO cells have been shown to bind human FcgRIIIa with a 50-fold increased affinity compared to those produced in wild-type CHO cells.
A NOVEL EXPRESSION SYSTEM Given the significance and impact of glycan engineering enhancement of mAbs for therapeutic use, Sino Biological has developed the FucoFree eukaryotic cell culture system using gene editing tech- niques. Tese FucoFree cell lines have undergone extensive testing, both at the
genetic and protein levels, to confirm the exhaustive knockout of the FUT8 gene and the absence of fucose residues in the antibody glycan chain.
Fig. 3. High-quality antibodies produced in FucoFree cell lines. A) Recombinant antibody demonstrating >95% purity and >98% monomer distribution. (B) LC- MS analysis of a mAb expressed in a conventional (two top panels in B) and the corresponding “FucoFree” (two bottom panels in B) cell line. A difference in the heavy chain molecular weight of 145 Da was observed, corresponding well with the MW of a fucose residue
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Host cells in the FucoFree system include a FUT8 knockout (FUT-/-) CHO cell line and a FUT-/- HEK293 cell line, each individually engineered to satisfy the demands of various projects and scale of production. Starting from amino acid sequences of antibody provided by customers, Sino Biological will deliver the afucosylated mAbs with its glycan profiling (Fig. 3B). Te company provides high-yield and high-throughput antibody expression with >95% purity and >98% monomer distribution (Fig. 3A). It can also assess its ADCC response and perform ADCC- enhanced mAb candidate screening. Te production scale can range from milligram to gram level. Afucosylated mAb expression is also included in the firm’s HTP services, rendering faster turnaround time and lower costs. A stable and high afucosylated mAb expression cell line can also be established. Currently, at least 35 glycoengineered antibodies, with their Fc fucose partially or entirely removed, have been investigated in animal models or clinical trials. Sino Biological anticipates that its services with the expression of afucosylated mAb will be helpful in the research and development community and open doors for more afucosylated mAb to enter clinical trials and subsequently be approved for use in humans.
Dr Emma Zhao is with Sino
Biological.www.sinobiological.com
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