PDF(Pubmed)
Abstract：
The manufacturability assessment and optimization of bispecific antibodies (bsAbs) during the discovery stage are crucial for the success of the drug development process, impacting the speed and cost of advancing such therapeutics to the Investigational New Drug (IND) stage and ultimately to the market. The complexity of bsAbs creates challenges in employing effective evaluation methods to detect developability risks in early discovery stage, and poses difficulties in identifying the root causes and implementing subsequent engineering solutions. This study presents a case of engineering a bsAb that displayed a normal solution appearance during the discovery phase but underwent significant precipitation when subjected to agitation stress during 15 L Chemistry, Manufacturing, and Control (CMC) production Leveraging analytical tools, structural analysis, in silico prediction, and wet-lab validations, the key molecular origins responsible for the observed precipitation were identified and addressed. Sequence engineering to reduce protein surface hydrophobicity and enhance conformational stability proved effective in resolving agitation-induced aggregation. The refined bsAb sequences enabled successful mass production in CMC department. The findings of this case study contribute to the understanding of the fundamental mechanism of agitation-induced aggregation and offer a potential protein engineering procedure for addressing similar issues in bsAb. Furthermore, this case study emphasizes the significance of a close partnership between Discovery and CMC teams. Integrating CMC\'s rigorous evaluation methods with Discovery\'s engineering capability can facilitate a streamlined development process for bsAb molecules.
摘要：
在发现阶段，双特异性抗体（bsAb）的可制造性评估和优化对于药物开发过程的成功至关重要。影响将此类疗法推进到研究新药（IND）阶段并最终推向市场的速度和成本。bsAbs的复杂性在早期发现阶段采用有效的评估方法来检测开发风险方面带来了挑战，并在确定根本原因和实施后续工程解决方案方面存在困难。本研究提供了一个工程bsAb的案例，该bsAb在发现阶段表现出正常的溶液外观，但在15L化学过程中受到搅拌应力时经历了明显的沉淀，Manufacturing,和控制(CMC)生产利用分析工具，结构分析,在硅预测中，和湿实验室验证，确定并解决了导致观察到的沉淀的关键分子起源。减少蛋白质表面疏水性和增强构象稳定性的序列工程被证明可有效解决搅拌诱导的聚集。精制的bsAb序列使CMC部门成功批量生产。本案例研究的发现有助于理解搅动诱导聚集的基本机制，并为解决bsAb中的类似问题提供了潜在的蛋白质工程程序。此外，本案例研究强调了Discovery和CMC团队之间紧密合作的重要性.将CMC的严格评估方法与Discovery的工程能力相结合，可以简化bsAb分子的开发过程。
