A Comparison of the Oligosaccharide Structures of Antithrombin Derived from Plasma and Recombinant Using POTELLIGENT® Technology

• Published in: Analytical Sciences Vol. 35; no. 12; pp. 1333 - 1340
• Main Authors: YAGI, Yuki, OKAZAKI, Akira, ENDO, Megumi, YANAGISAWA, Kumi, FUKUDA, Jun, NISHIMURA, Koichiro, YAMAZAKI, Katsuyoshi
• Format: Journal Article
• Language: English
• Published: Singapore The Japan Society for Analytical Chemistry 10.12.2019; Springer Nature Singapore; Japan Science and Technology Agency
• Subjects: Analytical Chemistry; Antithrombin; Antithrombins - chemistry; Antithrombins - metabolism; Chemistry; Fucose; Fucosyltransferases - deficiency; Fucosyltransferases - genetics; Gene Knockout Techniques; Genetic Engineering - methods; Glycan; glyco-engineering; Glycosylation; Humans; Isoforms; Mannose; oligosaccharide profile; Oligosaccharides; Oligosaccharides - chemistry; Plasma - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Safety; Technology; fucose; glyco-engineering; Antithrombin; oligosaccharide profile
• ISSN: 0910-6340; 1348-2246
• DOI: 10.2116/analsci.19P181

Human antithrombin (AT) has two isoforms of which the predominant α-form is glycosylated on all four possible glycosylation sites and the lower abundant β-isoform lacks the oligosaccharide on Asn135. The main oligosaccharide structure of human AT consists of biantennary complex-type oligosaccharides lacking a core fucose. Generally, Chinese hamster ovary (CHO) cells produce recombinant human AT (rhAT) with core-fucosylated oligosaccharides. However, rhAT lacking core-fucose oligosaccharides can be produced by POTELLIGENT® technology, which uses FUT8 knockout CHO cells in production. The rhAT has more variable glycan structures, such as tetra-antennary complex type, high-mannose type, and mannose 6-phosphate species as minor components compared to plasma-derived human AT (phAT). In addition, the site-specific glycan profile was different between two ATs. We evaluated the effect of these properties on efficacy and safety based on a comparison of rhAT made by that technology with phAT in terms of their respective oligosaccharide structures, site-specific oligosaccharide profiles, and the ratio of α- and β-forms. Although some structural differences were found between the rhAT and phAT, we concluded that these differences have no significant effect on the efficacy and safety of rhAT.