title: Glycosylation of human recombinant gonadotrophins: characterization and batch-to-batch consistency.
abstract: The glycan moiety of human recombinant gonadotrophins (r-hFSH, r-hLH, and r-hCG) produced in CHO cell lines has been characterized by a combination of chromatographic and mass spectrometric techniques, including both matrix-assisted laser desorption ionization and electrospray. Two glycan mapping methods have been developed for the three gonadotrophins that allow separation of the glycans according to either their charge or sialylation level or their antennarity. A method was also developed for r-hCG that permits the complete resolution of the N-glycan from the O-glycan species. Whereas the structure found for the N-glycans of the gonadotrophins was in agreement with the complex type model, the structure for an O-glycan of r-hCG, not yet described, has been unambiguously determined using nanoelectrospray ion trap mass spectrometry. Using these two glycan mapping methods, the high level of batch-to-batch consistency achieved for the glycosylation of the three recombinant gonadotrophins in commercial production has been shown. These data demonstrate the tight control that can be achieved in the manufacturing of complex recombinant therapeutic glycoproteins, which is a prerequisite to the delivering of a guaranteed dose of drug from vial to vial, and in turn to ensuring the clinical efficacy of the product.
authors.affiliation: Laboratoires Serono Sa, Serono Biotech Center, Zone Industrielle B, CH-1809 Fenil-Sur-corsier, Switzerland. annick.gervais@serono.com
authors.forename: Annick
authors.initials: A
authors.lastname: Gervais
authors.forename: Yves-Alexis
authors.initials: YA
authors.lastname: Hammel
authors.forename: Sophie
authors.initials: S
authors.lastname: Pelloux
authors.forename: Pierre
authors.initials: P
authors.lastname: Lepage
authors.forename: Gianni
authors.initials: G
authors.lastname: Baer
authors.forename: Nathalie
authors.initials: N
authors.lastname: Carte
authors.forename: Odile
authors.initials: O
authors.lastname: Sorokine
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Strub
authors.forename: Roman
authors.initials: R
authors.lastname: Koerner
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Leize
authors.forename: Alain
authors.initials: A
authors.lastname: Van Dorsselaer
mesh_terms: D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006224:Cricetinae; D006031:Glycosylation; D006062:Gonadotropins / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D055672:Static Electricity
chemical_list: D006062:Gonadotropins; D011134:Polysaccharides; D011994:Recombinant Proteins