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Glycosylation of human recombinant gonadotrophins: characterization and batch-to-batch consistency
- Source :
- Glycobiology. 13(3)
- Publication Year :
- 2003
-
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.
- Subjects :
- endocrine system
Glycan
Electrospray
Glycosylation
Molecular Sequence Data
Static Electricity
Molecular Conformation
Mass spectrometry
Biochemistry
law.invention
chemistry.chemical_compound
law
Polysaccharides
Cricetinae
Carbohydrate Conformation
Animals
Humans
Chromatography
biology
Chinese hamster ovary cell
Reproducibility of Results
Recombinant Proteins
carbohydrates (lipids)
Matrix-assisted laser desorption/ionization
chemistry
Carbohydrate Sequence
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
biology.protein
Recombinant DNA
Carbohydrate conformation
hormones, hormone substitutes, and hormone antagonists
Gonadotropins
Subjects
Details
- ISSN :
- 09596658
- Volume :
- 13
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Glycobiology
- Accession number :
- edsair.doi.dedup.....f9e6707af8ff6e49d803958939d5d483