Investigation of O-glycosylation heterogeneity of recombinant coagulation factor IX using LC–MS/MS. (September 2017)

• Record Type: Journal Article
• Authors: Seo, Youngsuk; Park, Gyeong Mi; Oh, Myung Jin; An, Hyun Joo
• Is Part Of: Bioanalysis. Volume 9:Number 18(2017)
• Page Start: 1361
• Page End: 1372
• Publication Date: 2017-09
• Subjects: O-glycosylation heterogeneity -- rFIX -- recombinant coagulation factor IX -- site-specific profiling; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2017-0086
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: Recombinant coagulation factor IX (rFIX) has extraordinarily multiple post-translational modifications including N -glycosylation and O -glycosylation which have a drastic effect on biological functions and in vivo recovery. Unlike N -glycosylation extensively characterized, there are a few studies on O -glycosylation due to its intrinsic complexity. In-depth O -glycosylation analysis is necessary to better understand and assess pharmacological activity of rFIX.Results: We determined unusual O -glycosylations including O -fucosylation and O -glucosylation which were located at Serine 53 and 61, respectively in EGF domain. Other O -glycosylations bearing core 1 glycan moiety were found on activation peptide.Conclusion: This is the first comprehensive study to characterize O -glycosylation of rFIX using MS-based glycomic and glycoproteomic approaches. Site-specific profiling will be a powerful platform to determine bioequivalence of biosimilars.