Novel Analytics for Rapid Adventitious Agent Detection in Cell and Gene Therapy Manufacturing

Commercial manufacturing of autologous cell and gene-modified cell therapies is logistically complex because of the requirement for significant coordination of clinical and manufacturing activities to ensure that the product and the patient are simultaneously prepared for administration. The final product must be held for 7-14 days pending a final negative result from growth-based compendial sterility tests, indicating product safety. Viral safety testing, which requires 28 days or longer, is uncommon. These timeframes are not optimal for cell therapy manufacturing because the patient’s condition may deteriorate, or final product quality may be impacted during this hold time. There is an unmet need for methods capable of rapid detection of a broad spectrum of adventitious agents (AAs), including the ability to distinguish between live/dead and infectious/non-infectious agents. In addition to allowing rapid release of cell therapy final products, rapid AA detection would also allow process monitoring to achieve early detection of contamination events which would impact the ability to supply the cell therapy product in the desired timeframe. 

We have developed multiple approaches to AA detection which can achieve the sensitivity of compendial sterility tests and the standard in vitro virus assay in much shorter timeframes. These include both targeted assays for detection of specific AA contaminants and untargeted approaches for broad spectrum detection of unknown contaminants. This talk provides an overview of methods that we have developed using multiple technology platforms such as the digital CRISPR-LAMP platform, next-generation sequencing using the Oxford Nanopore long-read sequencing platform, extracellular viral microRNA profiling, and metabolomic screening, which is being translated to both an at-line microfluidic mass spectrometry platform and a spectrophotometric platform which enables continuous, in-line cell culture monitoring.