[Feasibility evaluation of an ATP bioluminescence assay for rapid detection of microbial contamination in cell-based...

This study aims to investigate the feasibility of an ATP bioluminescence assay for rapid detection of microbial contamination in cell-based products. Firstly, we evaluated the applicability of a sample pre-treatment method for its ability to eliminate background ATP interference derived from animal cells. The effectiveness of the ATP bioluminescence assay for rapidly detecting microbial contamination was concurrently assessed. On this basis, the ATP bioluminescence assay combined with the pre-treatment method and the pharmacopeial sterility test were employed to analyze the animal cell samples spiked with microorganisms at low levels. The detection limits of both methods were determined, and the non-inferiority of the ATP bioluminescence assay relative to the pharmacopeial method was evaluated. The results showed that the pre-treatment method effectively eliminated the interference caused by background ATP signals from animal cells. The ATP bioluminescence assay detected microbial contamination rapidly. Compared with the pharmacopeial sterility test method, the ATP bioluminescence assay showed no statistically significant difference in detection limit and demonstrated non-inferiority in terms of decision equivalence. Therefore, within a risk-based strategy, the ATP bioluminescence assay is suitable for the rapid detection of microbial contamination in cell-based products. It provides rapid and effective contamination monitoring methods for the risk release of cell therapy drugs and the process control of aseptic processes for other biological products based on cell fermentation.