The glycosylation heterogeneity of recombinant human IFN-gamma.
The cloning of the cDNA for human interferon-gamma (IFN-gamma) has resulted in its expression in Escherichia coli, baculovirus-infected insect cells, Chinese hamster ovary (CHO) cells, and the mammary gland of transgenic mice. Large quantities of highly purified recombinant IFN-gamma have been generated, aided by the use of highly specific neutralizing monoclonal antibodies, with a view to its production as a human therapeutic protein. The primary source of structural heterogeneity for IFN-gamma during its production in mammalian expression systems is glycosylation, which can profoundly affect the three-dimensional structure of a glycoprotein and its biological function. A number of analytical approaches have been developed recently to allow a detailed analysis of the carbohydrate structures associated with IFN-gamma, the principal advances being in the areas of capillary electrophoresis and mass spectrometry. The implementation of these high-resolution analytical tools to determine the glycosylation profile of IFN-gamma makes it one of the best characterized recombinant glycoproteins. Recombinant human IFN-gamma acts as a model secretory glycoprotein, typifying the intrinsic glycosylation processing events associated with production of a potential therapeutic glycoprotein.
Hooker
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Oxford GlycoSciences (UK) Plc, Abingdon Science Park, United Kingdom. andrew.hooker@OGS.CO.UK
James
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D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D019075:Electrophoresis, Capillary / Q000379:methods; D006020:Glycopeptides / Q000032:analysis*; D006031:Glycosylation; D006801:Humans; D007371:Interferon-gamma / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D009844:Oligosaccharides / Q000032:analysis*; D011994:Recombinant Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006020:Glycopeptides; D009844:Oligosaccharides; D011994:Recombinant Proteins; D007371:Interferon-gamma