ATP生物发光方法应用于细胞类制品微生物污染快速检测的可行性评估

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

摘要:

本研究旨在探索三磷酸腺苷(adenosine triphosphate, ATP)生物发光快速微生物检测方法应用于细胞类制品污染检测的可行性。首先通过前处理方法适用性研究，评估前处理方法消除动物细胞背景ATP干扰的可行性，以及ATP生物发光法快速检出微生物污染的有效性。在此基础上采用结合前处理方法的ATP生物发光法以及药典无菌检查法对加标低浓度污染微生物的动物细胞供试品进行测试，统计2种方法的检测限，并评估ATP生物发光法相对于药典方法的非劣效性。结果显示，前处理方法可有效消除动物细胞背景ATP信号导致的干扰，同时ATP生物发光法可快速检出微生物污染。与药典无菌检查法相比，ATP生物发光法的检测限在统计学上无显著性差异，在决策等效性方面非劣效于药典方法。因此，基于风险评估，本研究中的ATP生物发光法可用于细胞类制品微生物污染的快速检测，为细胞治疗药物的风险放行以及基于细胞发酵的其他生物制品无菌工艺的过程控制提供了快速有效的污染监控手段。

Abstract:

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.