A Solution to Negate False Positives for Your Biologics
For years, only in vivo and in vitro assays were used to screen the cell banks, virus stocks, and serums used in production of biologics. Recent studies (in vitro/in vivo adventitious assays and bovine and porcine viruses) and contamination events have demonstrated the limitations of these assays. New approaches like PCR/qPCR and next generation sequencing (NGS) have been developed based on the detection of viral nucleic acids.
PCR Versus NGS
PCR is a useful alternative dealing with a small number of targets but remains challenging to implement for the detection of many targets. Moreover, PCR cannot detect new or emerging viruses because it relies on previous knowledge of the target.
NGS detects all nucleic acids present in a sample without prior knowledge of the sequence. The capability of NGS to detect new viruses has made this technology a top complemental test, and an alternative test for viral safety. A major concern about NGS is its potential to detect viral sequences that are not related to infectious viruses, resulting in a false positive and ultimately impacting the production process, resulting in lengthy and costly investigations.
Our partner, PathoQuest, has developed a specific NGS solution dedicated to viral safety assessment of cell banks (master and working cell bank, cell therapeutics, etc.) to improve the time to release and ease the manufacturing process. Through increased specificity and biological relevance, this unique approach reduces the number of false positives.
PathoQuest's transcriptome service differentiates inert carryover (irradiated BVDV-2) from a truly replicating virus (infectious BVDV-2) by ruling out the sequences coming from the genome of the RNA virus and those coming from the messenger RNA. Standard NGS only assesses the presence of RNA and DNA nucleic acids from cell lysate and/or supernatant, with a high risk of false positive hits, PathoQuest’s transcriptome NGS solution relies on the detection of the viral messenger RNA. The viral messenger RNAs that are an intermediate of replication synthesized only during viral infection by all virus types.
Why Use PathoQuest's NGS Solution?
- It has the capability of distinguishing the inert genome from the active messenger RNA, which is significant to eliminate the risk of false positives.
- It includes a proprietary data analysis pipeline, including a smart step of hit invalidation. The goal of this step is to determine the relevance of a hit and to classify this hit as a "positive hit" or a "background unspecific hit".
- It is fully validated according to industry standard guidelines including ICH Q2 (R1) and 21 CFR Part 11.
PathoQuest's validated Transcriptome NGS solution allows a broad-spectrum detection of all replicating infectious viruses (RNA and DNA viruses) with improved specificity and validated sensitivity compared to standard NGS. This method is the solution to replace in vivo assay screening of your master cell banks and accelerates the release of your cell therapeutics
Contact us to learn more about our NGS services and let's get your biologics to market faster.