Hong Wang
Shanghai Institute for Food and Drug Control, Shanghai, China
Title: Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine
Biography
Biography: Hong Wang
Abstract
Human follicle-stimulating hormone (hFSH) plays a key role in the development and function of the reproductive system. Two classes of hFSH-containing pharmaceutical preparations currently exist; those derived from the urine of postmenopausal women (uhFSH) and those manufactured using recombinant DNA technology (rhFSH). Comparative clinical studies have revealed the differences in oocyte quality and clinical outcome between rhFSH and uhFSH. The oligosaccharide moiety is critical in determining the pharmacological properties of therapeutic glycoproteins. hFSH is a glycoprotein and glycan significantly influences the biological properties of hFSH. Considering the importance of hFSH glycosylation to the biological activity of hFSH, evaluating the difference in glycosylation between rhFSH and uhFSH intended for clinical use is essential. Using a glycoproteomic strategy, this study compared the glycosylation of uhFSH with that of rhFSH. Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman−Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH.