Alston Alice, Bova Rebecca A, Hasson Bradley
Global Quality Control, Global Quality AstraZeneca, Liverpool, UK; AstraZeneca, 6 Renaissance Way, Liverpool, L24 9JW, UK.
NGS Operations, BioReliance® Testing Services, MilliporeSigma, Rockville, MD, USA.
Biologicals. 2025 May;90:101828. doi: 10.1016/j.biologicals.2025.101828. Epub 2025 Apr 2.
Next Generation Sequencing (NGS) has proven itself as a suitable replacement technology for traditional viral safety assessment assays in the manufacturing of complex biologics. Most notably, its incorporation into ICH Q5A(R2) in November 2023 endorses the technology platform as a suitable alternative to traditional in vivo, in vitro and PCR-based testing for adventitious viruses based on the risk assessment of the product and its context for use. In addition to the finalization of ICH Q5A(R2), a separate European Pharmacopoeia chapter Ph. Eur. 2.6.41 (High Throughput Sequencing for the Detection of Viral Extraneous Agents) is under review to further outline and provide guidance for the application and validation of NGS-based methodologies. Within the context of avian-based quadrivalent influenza vaccine manufacturing, NGS provides an alternative viral safety assessment method to traditional in vivo based testing models which are requirements for every lot manufactured. However, prior to implementation of NGS alternative methodology for commercial product testing, suitability of the method must be demonstrated within the context of the product through appropriate assay validation. In line with ICHQ5A (R2), 3.2.5.2 Next Generation Sequencing, non-targeted NGS can replace In vivo with broad virus detection for unknown or unexpected virus species without a head-to-head comparison. Therefore, complying with ICH Q5A (R2) and also AstraZeneca internal risk assessment for adventitious agents' detection, a comparability study directly comparing In vivo to NGS was not completed. This article summarizes the collaborative effort between AstraZeneca and MilliporeSigma to replace the in vivo adventitious virus test for Live Attenuated Influenza Vaccine (LAIV) with NGS for broad virus detection as part of a comprehensive virus testing strategy.
下一代测序（NGS）已证明自身是复杂生物制品生产中传统病毒安全性评估检测的合适替代技术。最值得注意的是，它于2023年11月被纳入ICH Q5A（R2），这认可了该技术平台可作为基于产品风险评估及其使用背景，替代传统体内、体外和基于PCR的检测偶然病毒的合适方法。除了ICH Q5A（R2）定稿外，欧洲药典单独的一章Ph. Eur. 2.6.41（用于检测病毒外源因子的高通量测序）正在审核中，以进一步概述并为基于NGS方法的应用和验证提供指导。在基于禽类的四价流感疫苗生产背景下，NGS为传统基于体内的检测模型提供了一种替代的病毒安全性评估方法，而传统检测模型是每批生产的要求。然而，在将NGS替代方法用于商业产品检测之前，必须通过适当的检测验证在产品背景下证明该方法的适用性。根据ICHQ5A（R2），3.2.5.2下一代测序，非靶向NGS可在无需直接对比的情况下，替代体内检测以广泛检测未知或意外的病毒种类。因此，遵循ICH Q5A（R2）以及阿斯利康对偶然因子检测的内部风险评估，未完成将体内检测与NGS直接对比的可比性研究。本文总结了阿斯利康和默克密理博之间的合作成果，即用NGS替代减毒活流感疫苗（LAIV）的体内偶然病毒检测，以进行广泛病毒检测，作为全面病毒检测策略的一部分。
