TY - JOUR
T1 - Effect of bench-scale culture conditions on murine IgG heterogeneity
AU - Marino, Maria
AU - Corti, Angelo
AU - Ippolito, Agostino
AU - Cassani, Giovanni
AU - Fassina, Giorgio
PY - 1997/4/5
Y1 - 1997/4/5
N2 - A stable murine hybridoma cell line, secreting IgG1 antibodies (7H3) against the soluble type I receptor for Tumor Necrosis Factor (sTNF-R1), was cultivated in two different bioreactor systems, a hollow fiber and a stirred tank fermentor, in order to evaluate the effect of culture conditions on antibody structural and functional heterogeneity. Conventional serum- supplemented and serum-free media were chosen for fermentation in stirred tank bioreactor, whereas only serum-supplemented media were used for hollow fiber cultivation. Extent of glycosylation, determined by lectin binding assays, and charge heterogeneity of murine monoclonal antibodies displayed relevant variations according to the fermentation system used. After complete sugars removal by N-glycosidase F treatment, charge heterogeneity were still observed suggesting the occurrence of additional modifications at the protein level. In vitro culture in serum-supplemented media carried out with the hollow fibre system led to higher productivity but greater antibody charge heterogeneity end differences in lectinbinding profile than cultivation in the stirred tank bioreactor. Results cumulatively indicated that hybridoma cultivation methods, but also cultivation time, influence antibody heterogeneity, both in the protein end sugar moieties.
AB - A stable murine hybridoma cell line, secreting IgG1 antibodies (7H3) against the soluble type I receptor for Tumor Necrosis Factor (sTNF-R1), was cultivated in two different bioreactor systems, a hollow fiber and a stirred tank fermentor, in order to evaluate the effect of culture conditions on antibody structural and functional heterogeneity. Conventional serum- supplemented and serum-free media were chosen for fermentation in stirred tank bioreactor, whereas only serum-supplemented media were used for hollow fiber cultivation. Extent of glycosylation, determined by lectin binding assays, and charge heterogeneity of murine monoclonal antibodies displayed relevant variations according to the fermentation system used. After complete sugars removal by N-glycosidase F treatment, charge heterogeneity were still observed suggesting the occurrence of additional modifications at the protein level. In vitro culture in serum-supplemented media carried out with the hollow fibre system led to higher productivity but greater antibody charge heterogeneity end differences in lectinbinding profile than cultivation in the stirred tank bioreactor. Results cumulatively indicated that hybridoma cultivation methods, but also cultivation time, influence antibody heterogeneity, both in the protein end sugar moieties.
KW - fermentation
KW - hybridoma cell culture
KW - MAb heterogeneity
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U2 - 10.1002/(SICI)1097-0290(19970405)54:1<17::AID-BIT2>3.0.CO;2-0
DO - 10.1002/(SICI)1097-0290(19970405)54:1<17::AID-BIT2>3.0.CO;2-0
M3 - Article
C2 - 18634068
AN - SCOPUS:0031553963
VL - 54
SP - 17
EP - 25
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
SN - 0006-3592
IS - 1
ER -