Hsu Tiffany, Talley Mary Jo, Yang Ping, Geiselhoeringer Angela, Yang Cindy, Gorla Aditya, Rahman M Julhasur, Silva Lindsey, Chen Dayue, Yang Bin
Purification, Microbiology, and Virology, Genentech, a Member of the Roche Group, South San Francisco, California, USA.
Biotechnol Prog. 2024 Nov-Dec;40(6):e3485. doi: 10.1002/btpr.3485. Epub 2024 Jul 25.
It is important to increase manufacturing speed to make medicines more widely available. One bottleneck for CHO-based drug substance release is the in vitro viral (IVV) cell-based assay on unprocessed bulk. To increase process speed, we evaluate the suitability of replacing the IVV cell-based assay with next-generation sequencing (NGS). First, we outline how NGS is currently used in the pharmaceutical industry, and how it may apply to CHO virus testing. Second, we examine CHO virus contamination history. Since prior virus contaminants can replicate in the production bioreactor, we perform a literature search and classify 159 viruses as high, medium, low, or unknown risk based on their ability to infect CHO cells. Overall, the risk of virus contamination during the CHO manufacturing process is low. Only six viruses were reported to have contaminated CHO bioprocesses over the past several decades, and were primarily caused by fetal bovine serum or cell culture components. These virus contamination events can be mitigated through limitation and control of raw materials, combined with virus testing and virus clearance technologies. The list of CHO infectious viruses provides a starting framework for virus safety risk assessment and NGS development. Furthermore, ICH Q5A (R2) includes NGS as a molecular method for adventitious agent testing, paving a path forward for modernizing CHO virus testing.
提高生产速度以使药物更广泛可得很重要。基于CHO的原料药放行的一个瓶颈是对未处理的原液进行基于体外病毒（IVV）细胞的检测。为了提高工艺速度，我们评估了用下一代测序（NGS）取代基于IVV细胞的检测的适用性。首先，我们概述了NGS目前在制药行业中的使用方式，以及它如何应用于CHO病毒检测。其次，我们研究了CHO病毒污染历史。由于先前的病毒污染物可在生产生物反应器中复制，我们进行了文献检索，并根据159种病毒感染CHO细胞的能力将其分类为高、中、低或未知风险。总体而言，CHO生产过程中病毒污染的风险较低。在过去几十年中，仅报告有六种病毒污染了CHO生物工艺，且主要是由胎牛血清或细胞培养成分引起的。通过限制和控制原材料，结合病毒检测和病毒清除技术，可以减轻这些病毒污染事件。CHO感染性病毒列表为病毒安全风险评估和NGS开发提供了一个起始框架。此外，ICH Q5A（R2）将NGS列为用于检测外源因子的分子方法，为CHO病毒检测的现代化铺平了道路。
