Ren Tingwei, Tan Zhijun, Ehamparanathan Vivekh, Lewandowski Angela, Ghose Sanchayita, Li Zheng Jian
Biologics Development, Global Product Development and Supply, Bristol-Myers Squibb, Devens, Massachusetts.
Biotechnol Bioeng. 2021 Aug;118(8):2829-2844. doi: 10.1002/bit.27790. Epub 2021 Jun 1.
Antibody disulfide bond reduction has been a challenging issue in monoclonal antibody manufacturing. It could lead to a decrease of product purity and failure to meet the targeted product profile and/or specifications. More importantly, disulfide bond reduction could also impact drug safety and efficacy. Scientists across the industry have been examining the root causes and developing mitigation strategies to address the challenge. In recent years, with the development of high titer mammalian cell culture processes to meet the rapidly growing demand for antibody biopharmaceuticals, disulfide bond reduction has been observed more frequently. Thus, it is necessary to continue evolving the disulfide reduction mitigation strategies and developing novel approaches to maintain high product quality. Additionally, in recent years as more complex molecules (such as bispecific and trispecific antibodies) emerge, the molecular heterogeneity due to incomplete formation of the interchain disulfide bonds becomes a more imperative challenging issue. Given the disulfide reduction challenges that biotech industry is facing, in this review, we provide a comprehensive scientific summary of the root cause analysis of disulfide reduction during process development of antibody therapeutics, mitigation strategies and its potential remediated recovery based on published papers. First, this paper intends to highlight different aspects of the root cause for disulfide reduction. Secondly, to provide a broader understanding of the disulfide bond reduction in downstream process, this paper discusses disulfide bond reduction impact on product stability, associated analytical methods for disulfide bond reduction detection and characterization, process control strategies as well as their manufacturing implementation. In addition, brief perspectives on the development of future mitigation strategies are also reviewed, including platform alignment, mitigation strategy application for the emerging new modalities such as bispecific and trispecific antibodies as well as using machine learning to identify molecule susceptibility of disulfide bond reduction. The data in this review are originated from the published papers.
抗体二硫键还原一直是单克隆抗体制备过程中的一个具有挑战性的问题。它可能导致产品纯度下降，无法达到目标产品特性和/或规格要求。更重要的是，二硫键还原还可能影响药物的安全性和有效性。整个行业的科学家们一直在研究根本原因，并制定缓解策略来应对这一挑战。近年来，随着高滴度哺乳动物细胞培养工艺的发展，以满足对抗体生物制药迅速增长的需求，二硫键还原现象越来越频繁地被观察到。因此，有必要不断改进二硫键还原缓解策略，并开发新方法以保持高产品质量。此外，近年来随着更复杂的分子（如双特异性和三特异性抗体）出现，由于链间二硫键形成不完全导致的分子异质性成为一个更紧迫的挑战性问题。鉴于生物技术行业面临的二硫键还原挑战，在本综述中，我们基于已发表的论文，对抗体治疗药物工艺开发过程中二硫键还原的根本原因分析、缓解策略及其潜在的补救回收方法进行了全面的科学总结。首先，本文旨在突出二硫键还原根本原因的不同方面。其次，为了更广泛地理解下游工艺中的二硫键还原，本文讨论了二硫键还原对产品稳定性的影响、用于二硫键还原检测和表征的相关分析方法、工艺控制策略及其在生产中的实施。此外，还综述了未来缓解策略发展的简要观点，包括平台整合、双特异性和三特异性抗体等新兴新形式的缓解策略应用以及使用机器学习来识别二硫键还原的分子敏感性。本综述中的数据源自已发表的论文。
