Wang Shuang, Zhang Weijie, Yang Baotian, Zhang Xudong, Fang Jing, Rui Haopeng, Chen Zhijian, Gu Jijie, Chen Zhiqiang, Xu Jianqing
Biologics Innovation Discovery, WuXi Biologics, 1951 Huifeng West Road, Fengxian District, Shanghai, 201400, China.
Downstream Process Development (DSPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.
Antib Ther. 2024 May 29;7(3):189-198. doi: 10.1093/abt/tbae013. eCollection 2024 Jul.
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, 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.
双特异性抗体（bsAb）在发现阶段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，会影响将此类治疗药物推进到研究性新药（IND）阶段并最终推向市场的速度和成本。bsAb的复杂性给在早期发现阶段采用有效的评估方法来检测可开发性风险带来了挑战，并且在确定根本原因和实施后续工程解决方案方面也存在困难。本研究展示了一个双特异性抗体工程改造的案例，该抗体在发现阶段溶液外观正常，但在15L化学、制造和控制（CMC）生产过程中受到搅拌应力时会发生显著沉淀。通过利用分析工具、结构分析、预测和湿实验室验证，确定并解决了导致观察到的沉淀的关键分子根源。通过序列工程降低蛋白质表面疏水性并增强构象稳定性，被证明对解决搅拌诱导的聚集有效。优化后的bsAb序列使得在CMC部门能够成功进行大规模生产。本案例研究的结果有助于理解搅拌诱导聚集的基本机制，并为解决bsAb中类似问题提供了一种潜在的蛋白质工程方法。此外，本案例研究强调了发现团队和CMC团队紧密合作的重要性。将CMC严格的评估方法与发现团队的工程能力相结合，可以促进bsAb分子的简化开发过程。
