A structured approach for the evaluation, validation and implementation of NAT-based mycoplasma detection methods

Alternative adventitious agent detection methods, or adventitious agent tests (AATs) have been available for many years and use of alternative AATs has been advocated by global regulatory authorities, but widespread adoption in the biotherapeutic industry has been slow. Many traditional compendial methods used for the

detection of adventitious agents are laborious, difficult to automate to alleviate data integrity concerns, and can cause significant delays before results are obtained.

Previous work conducted by this forum¹ identified that reluctance to embrace these technologies, both existing and in development, could be attributed to breakdowns in:

Strategy and approach (e.g. unclear drivers, selection of inappropriate technology for the application)
Technical (e.g. regulatory acceptance, validation challenges)
Cultural (e.g. reservations around change technologies)
Implementation and execution (e.g. inadequate validation strategy, difficulties of global regulatory filings).

The BioPhorum biomanufacturing technology roadmap outlined the aspiration for a <2-day AAT for product release by 2026. However, as the biomanufacturing landscape has evolved to include advanced therapy medicinal products (ATMPs), many of which have a short shelf life, there is a clear need to embrace new technologies which provide an assurance of safety from adventitious agents in a much shorter timeframe. In addition, the global COVID-19 pandemic has emphasized an urgent need for changing paradigms to enable faster lot release and access to life-saving therapeutics.

To address some of the challenges identified as roadblocks to implementation of alternative AATs, this paper provides case studies and best practice guidance for validation of an alternative mycoplasma detection assay. It includes an example of completing a user requirement specification (URS) for evaluating an alternative mycoplasma detection assay, validation and implementation. Strategies are also provided to help identify suitable technologies for defined applications. Finally, a framework is given to demonstrate how a validation protocol for an alternative mycoplasma detection assay can be executed to meet the regulatory expectation that the alternative method is equivalent to or better than the current compendial methods (culture method and indicator cell culture method).