The biopharm industry commonly uses CHO cells to produce large quantities of recombinant monoclonal antibodies (mAbs). At the end of the culture process, cells are separated from the culture media during the primary recovery step using methods such as centrifugation, depth filtration, or flocculation to clarify the harvest fluid (clarified bulk or CB). The CB is further processed to downstream purification steps. There have been multiple observations, including ours, that reduction of mAb disulfide bonds is observed during primary recovery to downstream of the process. The reduction events mainly happen at scale-up manufacturing and may result in loss of whole batch of production due to failure to meet specifications. Many studies from different biopharm firms have enabled understanding of the molecular and cellular mechanisms of the reduction and advanced mitigation strategies to improve in-process control to ensure product quality. The relationship of oxygen consumption in CB in closed containers and antibody reduction is a key principle to develop effective mitigation strategies, such as air sparging. At Bristol-Myers Squibb, we have developed a systematic plan, including forecasting, monitoring, mitigation and rescue, to handle the antibody reduction issue. Using optimized concentrations of hydrogen peroxide or other peroxides can effectively prevent antibody reduction and extend CB holding time. The treatment does not impact on product quality attributes. Small-scale runs demonstrate that hydrogen peroxide in CB does not impact the performance of protein A chromatography step. Since hydrogen peroxide is ready to mix with water and its decomposition products are water and oxygen, it is expected no burden in downstream purification step to remove excess hydrogen peroxide. The simple treatment promises reduced costs, improved manufacturing capabilities and increased manufacturing flexibility.

Using hydrogen peroxide and derivatives to prevent antibody disulfide bond reduction during manufacturing process

2019 Spring National Meeting

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