Validation of a Next Generation Sequencing Method for adventitious agents detection in a live vaccine matrix

Abstract

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.