title: Process monitoring of the production of gamma-interferon in recombinant Chinese hamster ovary cells.
abstract: The production of recombinant gamma-interferon was monitored using high-performance liquid chromatographic methods. These methods were able to distinguish between glycosylated and non-glycosylated forms of gamma-interferon by complexing the carbohydrate with borate. Sufficient quantities of standard glycosylated gamma-interferon were not available for peak identification so immunological techniques were used to identify gamma-interferon variants. These techniques were validated with the non-glycosylated form. The non-glycosylated form was then shown to be retained only on a cation-exchange column, while the glycosylated form, complexed with borate, was retained only on an anion-exchange column. Samples were drawn at 2-h intervals over a 60-h production cycle and analyzed by both anion- and cation-exchange chromatography. Results indicated that the production of each form was coincidental and that the glycosylated form of gamma-interferon is produced in greater abundance than non-glycosylated.
authors.affiliation: Chemistry Department, Purdue University, West Lafayette, IN 47907.
authors.forename: T K
authors.initials: TK
authors.lastname: Nadler
authors.forename: S K
authors.initials: SK
authors.lastname: Paliwal
authors.forename: F E
authors.initials: FE
authors.lastname: Regnier
authors.forename: R
authors.initials: R
authors.lastname: Singhvi
authors.forename: D I
authors.initials: DI
authors.lastname: Wang
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006224:Cricetinae; D007371:Interferon-gamma / Q000378:metabolism*; D011994:Recombinant Proteins; D013997:Time Factors
chemical_list: D011994:Recombinant Proteins; D007371:Interferon-gamma