Rayfield William J, Roush David J, Chmielowski Rebecca A, Tugcu Nihal, Barakat Shehab, Cheung Jason K
Process Development and Engineering, Biologics Process Development, Merck Research Laboratories, Kenilworth, NJ, 07033.
Sterile Product and Analytical Development, Biologics Process Development, Merck Research Laboratories, Kenilworth, NJ, 07033.
Biotechnol Prog. 2015 May-Jun;31(3):765-74. doi: 10.1002/btpr.2094. Epub 2015 May 19.
Controlling viral contamination is an important issue in the process development of monoclonal antibodies (MAbs) produced from mammalian cell lines. Virus filtration (VF) has been demonstrated to be a robust and effective clearance step which can provide ≥4 logs of reduction via size exclusion. The minimization of VF area by increasing flux and filter loading is critical to achieving cost targets as VFs are single use and often represent up to 10% of total purification costs. The research presented in this publication describes a development strategy focused on biophysical attributes of product streams that are directly applicable to VF process performance. This article summarizes a case study where biophysical tools (high-pressure size exclusion chromatography, dynamic light scattering, and absolute size exclusion chromatography) were applied to a specific MAb program to illustrate how changes in feed composition (pH, sodium chloride concentration, and buffer salt type) can change biophysical properties which correlate with VF performance. The approach was subsequently refined and expanded over the course of development of three MAbs where performance metrics (i.e., loading and flux) were evaluated for two specific virus filters (Viresolve Pro and Planova 20N) during both unspiked control runs and virus clearance experiments. The analyses of feed attributes can be applied to a decision tree to guide the recommendation of a VF filter and operating conditions for use in future MAb program development. The understanding of the biophysical properties of the feed can be correlated to virus filter performance to significantly reduce the mass of product, time, and costs associated with virus filter step development.
控制病毒污染是哺乳动物细胞系生产单克隆抗体（MAb）工艺开发中的一个重要问题。病毒过滤（VF）已被证明是一个强大且有效的清除步骤，可通过尺寸排阻实现≥4个对数级的病毒减少。由于病毒过滤器是一次性使用的，且通常占总纯化成本的10%，因此通过提高通量和过滤器载量来最小化病毒过滤面积对于实现成本目标至关重要。本出版物中介绍的研究描述了一种开发策略，该策略侧重于直接适用于病毒过滤工艺性能的产品流生物物理属性。本文总结了一个案例研究，其中将生物物理工具（高压尺寸排阻色谱、动态光散射和绝对尺寸排阻色谱）应用于一个特定的单克隆抗体项目，以说明进料组成（pH值、氯化钠浓度和缓冲盐类型）的变化如何改变与病毒过滤性能相关的生物物理性质。在三种单克隆抗体的开发过程中，该方法随后得到了完善和扩展，在未加样的对照运行和病毒清除实验中，对两种特定的病毒过滤器（Viresolve Pro和Planova 20N）评估了性能指标（即载量和通量）。进料属性分析可应用于决策树，以指导未来单克隆抗体项目开发中病毒过滤过滤器和操作条件的推荐。对进料生物物理性质的理解可与病毒过滤器性能相关联，从而显著减少与病毒过滤步骤开发相关的产品质量、时间和成本。
