Process monitoring of the production of gamma-interferon in recombinant Chinese hamster ovary cells.
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.
Nadler
T K
TK
Chemistry Department, Purdue University, West Lafayette, IN 47907.
Paliwal
S K
SK
Regnier
F E
FE
Singhvi
R
R
Wang
D I
DI
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
D011994:Recombinant Proteins; D007371:Interferon-gamma