Report of the second international conference on next generation sequencing for adventitious virus detection in biologics for humans and animals

Abstract

The IABS-EU, in association with PROVAXS and Ghent University, hosted the “2 nd Conference on Next Generation Sequencing (NGS) for Adventitious Virus Detection in Human and Veterinary Biologics” held on November 13 th and 14 th 2019, in Ghent, Belgium. The meeting brought together international experts from regulatory agencies, the biotherapeutics and biologics industries, contract research organizations, and academia, with the goal to develop a scientific consensus on the readiness of NGS for detecting adventitious viruses, and on the use of this technology to supplement or replace/substitute the currently used assays. Participants discussed the progress on the standardization and validation of the technical and bioinformatics steps in NGS for characterization and safety evaluation of biologics, including human and animal vaccines. It was concluded that NGS can be used for the detection of a broad range of viruses, including novel viruses, and therefore can complement, supplement or even replace some of the conventional adventitious virus detection assays. Furthermore, the development of reference viral standards, complete and correctly annotated viral databases, and protocols for the validation and follow-up investigations of NGS signals is necessary to enable broader use of NGS. An international collaborative effort, involving regulatory authorities, industry, academia, and other stakeholders is ongoing toward this goal.

Advances for the Hepatitis A Virus Antigen Production Using a Virus Strain With Codon Frequency Optimization Adjustments in Specific Locations

“…Additionally, it has been reported to be free of tumorigenic features (Dotti et al, 2017) and of polyomavirus (Provost et al, 1984), the adventitious agent that prevented the use of the fetal rhesus cell lines for vaccine production (Klug et al, 2016). Moreover, Next-Generation-Sequencing (NGS) technologies for the detection of a broad range of viruses in combination with complete and correctly annotated viral databases may expand the range of cell substrates useful for vaccine production (Khan et al, 2020).…”

Attitude and perception survey for the Japanese pharmaceutical industry to utilize next-generation sequencing for virus safety assessment of biologics

“…We surveyed respondents' opinions on the necessity of virus-detection assays using animals, such as in vivo virus tests and antibody-production tests, and their replacement with molecular-based methods (Fig. 3), in light of the growing international discussion on the replacement of animal tests with NGS [5,16]. Currently, none of the respondents have any experience with NGS-based virus safety assessment with cell banks (Question 4-1).…”

“…Particularly in Europe, regulatory efforts toward 3Rs compliance are active and included in one of the regulatory science efforts by the European Medicines Agency [3]. The requirements for replacing the in vivo virus test with NGS or a related molecular-based method are also included in the European Pharmacopoeia [4,5].…”

“…In this context, since around 2012, mainly in Europe and the United States, industry, academia, and regulatory agencies have collaborated to implement NGS-based virus safety assessments [5][6][7][8][9][10]. In these activities, participants discussed technical considerations [11,12], conducted multicenter studies to assess the performance of NGS-based testing [5,13], and developed a reference viral genome database and standard reference viral reagents for spiking tests [5,14,15], all of which could provide specific images of the NGSbased virus safety assessment in discussions for ICH Q5A revision. However, it is currently unclear whether Japanese pharmaceutical companies are participating in such international initiatives, and it is also unclear whether they intend to use NGS for virus safety assessment.…”

k -mer-Based Metagenomics Tools Provide a Fast and Sensitive Approach for the Detection of Viral Contaminants in Biopharmaceutical and Vaccine Manufacturing Applications Using Next-Generation Sequencing

“…Second, an alternate method, PCR-based identification, requires the design of primers for a specific type of virus before it can be identified, and therefore, this method requires prior knowledge of possible contaminants (1,18). To overcome these challenges, next-generation sequencing (NGS) methods have recently been proposed as an alternative virus detection strategy (19)(20)(21)(22)(23). NGS can be much faster, with the sequencing only taking 1 to 2 days, although the sample processing and bioinformatics analysis can take anywhere from a few hours to weeks depending on what methods are used.…”