Glycosylation of human recombinant gonadotrophins: characterization and batch-to-batch consistency.
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.
Gervais
Annick
A
Laboratoires Serono Sa, Serono Biotech Center, Zone Industrielle B, CH-1809 Fenil-Sur-corsier, Switzerland. annick.gervais@serono.com
Hammel
Yves-Alexis
YA
Pelloux
Sophie
S
Lepage
Pierre
P
Baer
Gianni
G
Carte
Nathalie
N
Sorokine
Odile
O
Strub
Jean-Marc
JM
Koerner
Roman
R
Leize
Emmanuelle
E
Van Dorsselaer
Alain
A
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
D006062:Gonadotropins; D011134:Polysaccharides; D011994:Recombinant Proteins