Rapid Robust Mycoplasma Detection System for Release Testing of Cell and Gene Therapy Products Containing Mammalian Cells in 1-Hour

Introduction

Cell and gene therapies (C&GT) or Advanced Therapeutic Medicinal Products (ATMP) are innovative medicine developed to treat cancers, rare diseases, autoimmune disorders, and injuries. As these therapies utilize living cells, they produce a product with a short shelf life. While microbiological examination of cell-based products is critical to ensure patient safety, challenges exist to enable testing and obtaining rapid results prior to patient infusion.

One of these microbiological examinations is the mycoplasma testing of cellular therapy products which is a regulatory requirement for release. The compendial microbiological assay, currently recommended by the United States Pharmacopeia (USP), European Pharmacopeia (EP), Japanese Pharmacopeia (JP) and the US FDA, for mycoplasma testing of biologics, involves the culture of viable mycoplasmas in broth, agar plates and indicator cell cultures. This compendial culture testing utilizes complex media and requires ≥ 28 days to generate test results. Waiting 28 days or longer to release product isn’t a feasible option for most autologous cell and gene therapy processes where patients are waiting to be infused and time is critical to effective treatment. (1-3)

Conventional nucleic acid testing (NAT) methods offer a faster alternative to compendial methods with results often within a single work shift; however, they remain highly manual, require specialized laboratories, elevated skill levels and significant training to execute the test and interpret results. Because of these complexities, mycoplasma testing is often outsourced to third-party laboratories, resulting in additional costs.

BIOFIRE® Mycoplasma: BioMérieux provides an innovative test for mycoplasma detection in cell and gene therapy products. The BIOFIRE® Mycoplasma test can detect the presence of >130 species of mycoplasma. The system provides sample to answer in ~1 hour with little technical training required. This provides an easy-to-learn, easy-to-use option to bring mycoplasma testing in-house, saving time and outsourcing costs.

The objective is to Introduce C&GT product specific protocols that allows the inclusion of mammalian cells thus aligning with the upcoming EP guideline (4) whilst providing the required level of detection (≤ 10 CFU/mL) as a Mycoplasma release test.