Advancements in viral detection methods are set to transform the biologics industry, addressing critical challenges associated with viral contamination during production. Traditional techniques, while established, often fall short in sensitivity and speed, especially as the complexity of biologic products increases. In response, Next-Generation Sequencing (NGS) is emerging as a powerful alternative, offering faster and more comprehensive detection of viral pathogens.
Traditional methods for viral safety testing in biologics can be cumbersome and time-consuming. These include various in vivo and in vitro assays, such as animal adventitious virus assays and PCR, which can take weeks to complete. The recently revised ICH Q5A guideline highlights the need for improved viral safety measures and supports the integration of NGS into testing protocols, emphasizing its enhanced capabilities in detecting both known and unknown viruses.
As Dr. Horst Ruppach, Executive Director of Scientific and Portfolio at Charles River Laboratories, notes, “Simply use NGS but maybe not initially in a critical submission. In new product development, use it as an alternative or a supplement to the standard testing.” This approach allows companies to familiarize themselves with the technology while benefiting from its superior capabilities.
Advantages of Next-Generation Sequencing
Next-Generation Sequencing operates without bias, detecting all nucleic acid molecules within a sample. Its ability to identify viral genomes in diverse biological matrices sets it apart from traditional methods. NGS can analyze viral presence in cell-free products through virome analysis, or assess endogenous viruses in cellular genomes via genome analysis. The ability to deliver results within two to three weeks significantly reduces the turnaround time, a crucial factor in the fast-paced biologics market.
Cell lines such as CHO, Vero, and HEK293 are standard in the production of biologics like vaccines and recombinant proteins. Comprehensive screening for replication-competent viruses is essential, as even low-level contamination can proliferate during cell culture. NGS offers a more thorough approach by detecting all types of viral genomes and variants, making it a vital tool for ensuring product safety.
Despite its advantages, the complexity of the NGS technology poses challenges, particularly in developing and interpreting bioinformatics pipelines. Collaborative efforts, such as those between Charles River Laboratories and PathoQuest, aim to refine these processes, enhancing the reliability of viral detection.
Implications for Cell Therapy and Future Outlook
The rise of cell therapy products, particularly allogeneic therapies, introduces unique viral safety challenges. Unlike traditional biologics, these therapies involve the direct administration of cells to patients, leaving no opportunity for the removal of viral contaminants. The rigorous testing required for master virus seeds and viral product batches often struggles under traditional methods, which may not detect certain contaminants.
Dr. Ruppach emphasizes the necessity of adopting NGS for future viral safety testing, stating, “In the future, it will be mandatory to use NGS because it has the highest breadth of detection.” The technology’s ability to identify viral mRNA directly confirms the presence of active viruses, offering a significant advantage over conventional assays.
Recent studies demonstrate NGS’s superiority over traditional methods. For example, side-by-side tests conducted by Charles River Laboratories and PathoQuest showed that NGS not only matched sensitivity but also surpassed it in numerous instances, identifying previously undetected viruses.
With regulatory bodies increasingly recognizing the value of NGS, the shift towards this technology is gaining momentum. Improved workflows in sample preparation and analysis are streamlining processes, allowing for compliance-ready results with reduced labor. NGS further aligns with global ethical testing trends, as it minimizes reliance on animal testing.
Looking ahead, Dr. Ruppach predicts that NGS will become standard practice in viral testing within five years. “In five years, people may wonder why we used the traditional approaches when we had this technology,” he concluded. As the biologics industry continues to evolve, the integration of NGS into viral detection protocols promises to enhance safety and efficiency significantly.
