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Air sparging for prevention of antibody disulfide bond reduction in harvested CHO cell culture fluid
Author(s) -
Mun Melissa,
Khoo Stefanie,
Do Minh Aline,
Dvornicky James,
TrexlerSchmidt Melody,
Kao YungHsiang,
Laird Michael W.
Publication year - 2015
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25495
Subject(s) - chinese hamster ovary cell , sparging , air sparging , bioreactor , antibody , cell culture , chemistry , lysis , chromatography , biochemistry , biology , immunology , organic chemistry , contamination , environmental remediation , ecology , genetics , receptor
During the scale‐up of several Chinese Hamster Ovary (CHO) cell monoclonal antibody production processes, significant reduction of the antibody interchain disulfide bonds was observed. The reduction was correlated with excessive mechanical cell shear during the harvest operations. These antibody reduction events resulted in failed product specifications and the subsequent loss of the drug substance batches. Several methods were recently developed to prevent antibody reduction, including modifying the cell culture media, using pre‐ and post‐harvest chemical additions to the cell culture fluid (CCF), lowering the pH, and air sparging of the harvested CCF (HCCF). The work described in this paper further explores the option of HCCF air sparging for preventing antibody reduction. Here, a small‐scale model was developed using a 3‐L bioreactor to mimic the conditions of a manufacturing‐scale harvest vessel and was subsequently employed to evaluate several air sparging strategies. In addition, these studies enabled further understanding of the relationships between cell lysis levels, oxygen consumption, and antibody reduction. Finally, the effectiveness of air sparging for several CHO cell lines and the potential impact on product quality were assessed to demonstrate that air sparging is an effective method in preventing antibody reduction. Biotechnol. Bioeng. 2015;112: 734–742. © 2014 Wiley Periodicals, Inc.