title: Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine.
abstract: Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 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. The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH.
authors.affiliation: Shanghai Institute for Food and Drug Control , Shanghai 201203, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Waters Corporation , Shanghai 201206, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Thermo Fisher Scientific , Shanghai 201206, China.
authors.forename: Xiaoxi
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Thermo Fisher Scientific , Shanghai 201206, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Shanghai Techwell Biopharmaceutical Corporation , Shanghai 201108, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Shanghai Institute for Food and Drug Control , Shanghai 201203, China.
authors.forename: Hongrui
authors.initials: H
authors.lastname: Yin
authors.affiliation: Shanghai Institute for Food and Drug Control , Shanghai 201203, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Shao
authors.affiliation: Shanghai Institute for Food and Drug Control , Shanghai 201203, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006224:Cricetinae; D003412:Cricetulus; D043373:Follicle Stimulating Hormone, Human / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D006031:Glycosylation; D006801:Humans; D008970:Molecular Weight; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D043373:Follicle Stimulating Hormone, Human; D011134:Polysaccharides; D011994:Recombinant Proteins
keywords: follicle-stimulating hormone; glycosylation; mass spectrometry; rhFSH; uhFSH