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Investigation of the free heavy chain homodimers of a monoclonal antibody

机译：单克隆抗体自由重链同源体的研究

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摘要

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 (R-20.9) 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 retransfecting the MTX amplified pools with additional LC gene. (c) 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:738-745, 2018
机译：单克隆抗体（mAb）由两种重链（HC）和两种轻链（LC）多肽组成。 HC和LC的适当折叠和组装对于抗体生产至关重要。目前的教条表明，除非用LCS与抗体组装成抗体，否则目前的RCS保留在内质网（ER）中，而另一方面的LCS可以分泌为游离单体或二聚体分子。在该研究中，在MAB1的细胞系发育期间，在细胞培养基中观察到高水平的细胞外HC同源体（7％-45％）。游离HC同源体水平与高分子量物质的百分比之间的优异相关性（R-2> 0.9）表明游离HC同型二聚体可能是造成不希望的聚集的。由于HC同源体的表面电荷不同和完全组装的抗体，通过阳离子交换色谱步骤成功地除去不需要的细胞外HC同源体。降低来自不同MTX扩增池的细胞培养基（RCE-SDS）分析的减少的毛细管电泳 - 硫酸钠（RCE-SDS）分析表明，LC表达不充分是高水平的游离HC同态二聚体的一个潜在根本原因。用另外的LC基因重传MTX扩增池后，游离HC同源体的水平显着（3％-25％）在缩回后。 （c）2018美国化学工程师学院Biotechnol。 Prog。，34：738-745,2018

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