In vivo glyco‐engineered antibody with improved lytic potential produced by an innovative non‐mammalian expression system

Abstract

Recent studies have demonstrated that the reduction of the core fucosylation on N-glycans of human IgGs is responsible for a clearly enhanced antibody-dependent cellular cytotoxicity (ADCC). This finding might give access to improved active therapeutic antibodies. Here, the expression of the tumor antigen-specific antibody IGN311 was performed in a glyco-optimized strain of the moss Physcomitrella patens. Removal of plant specific N-glycan structures in this plant expression host was achieved by targeted knockout of corresponding genes and included quantitative elimination of core fucosylation. Antibodies transiently expressed and secreted by such genetically modified moss protoplasts assembled correctly, showed an unaltered antigen-binding affinity and, in extensive tests, revealed an up to 40-fold enhanced ADCC. Thus, the glyco-engineered moss-based transient expression platform combines a rapid technology with the subsequent analysis of glycooptimized therapeutics with regard to advanced properties.

Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

“…The heavy chain from the protein A-purified antibody contained the expected, correctly processed N-terminal peptide EVQLVESGGGLVK, which was absent from the degradation product. The full-size heavy chain partially lacked the C-terminal lysine residue, as was recently found in the IGN314 antibody produced in moss (32), whereas this residue was absent from most of the degradation product. The continuous peptide map of the degradation product ran from L123 to K472, with the proteolytic cleavage site presumably located between G119 and L123.…”

EB66 cell line, a duck embryonic stem cell-derived substrate for the industrial production of therapeutic monoclonal antibodies with enhanced ADCC activity

“…[28][29][30][31][32] Some cells naturally produce proteins with lower fucose content. The critical role of the FUT-8 gene in fucosylation and the correlation between low FUT-8 expression and higher ADCC activity of the produced antibodies was thus initially reported for the rat hybridoma Y2B/0 cell line.…”

“…[28][29][30][31][32] While these systems may offer attractive potential advantages such as cost-effectiveness compared with mammalian cell cultures and a reduced risk of the presence of animal adventitious agents, they also present some natural drawbacks such as the presence of non-human oligosaccharides that may elicit in humans an immune response and alter the pharmacokinetics of the antibodies. Humanization of such yeast or plant production hosts by glycoengineering, i.e., elimination of endogenous genes responsible for undesired glycosylations and introduction of human genes controlling the preferred Figure 8.…”

Production of therapeutic antibodies with controlled fucosylation

“…67 None of the products from the engineered moss, vascular endothelial growth factor (VEGF), IgG4, and IgG1, are reported to carry either α-1,3 fucose or β-1,2 xylose on N-glycans. [67][68][69] The double gene knockdown approach has been also performed in duckweed. 70 The engineered duckweed expressed IgG1 with almost total elimination of α-1,3 fucose and β-1,2 xylose on N-glycans (95.8%).…”