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Investigation of the Free Heavy Chain Homodimers of a Monoclonal Antibody

Abstract

Monoclonal antibodies (mAbs) are composed of two heavy chain (HC) and two light chain (LC) polypeptides. The proper folding and assembly of HC and LC is critical for antibody production. Current dogma indicates that the free HCs are retained in the endoplasmic reticulum (ER) unless assembled with LCs into antibodies, while the LCs on the other hand can be secreted as free monomer or dimer molecules. In this study, high levels of extracellular HC homodimers (7–45%) were observed in the cell culture media during cell line development for mAb1. Excellent correlation (R2>0.9) between the level of free HC homodimers and the percentage of high molecular weight species indicates that the free HC homodimers might be causative of unwanted aggregation. Due to the different surface charge of HC homodimer and fully assembled antibodies, the unwanted extracellular HC homodimers were successfully removed by downstream processing, through a cation exchange chromatography step. Reduced capillary electrophoresis‐sodium dodecyl sulfate (rCE‐SDS) analysis of the cell culture media from different MTX amplified pools indicated that insufficient expression of LC is one potential root cause for the high level of free HC homodimers. The level of free HC homodimers decreased significantly (3–25%) after re‐transfecting the MTX amplified pools with additional LC gene. This article is protected by copyright. All rights reserved.

Authors:   Hyo (Helen) Chung, Lynette Buck, Kristi Daris, Brent Welborn, Quanzhou Luo, Jette Wypych
Journal:   Biotechnology Progress
Year:   2018
Pages:   n/a
DOI:   10.1002/btpr.2611
Publication date:   17-Jan-2018
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