Effect of bench-scale culture conditions on murine IgG heterogeneity

Maria Marino, Angelo Corti, Agostino Ippolito, Giovanni Cassani, Giorgio Fassina

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)17-25
Number of pages9
JournalBiotechnology and Bioengineering
Volume54
Issue number1
DOIs
Publication statusPublished - Apr 5 1997

Keywords

  • fermentation
  • hybridoma cell culture
  • MAb heterogeneity

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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