【摘要】 目的:旨在建立高效、可靠的实验方法以快速检测肿瘤浸润淋巴细胞（tumor-infiltrating lymphocyte, TIL）产品制备过程中多种细胞培养基辅料的潜在残留。方法:采用酶联免疫吸附法（enzyme-linked immunosorbent assays, ELISA）定量检测TIL产品中的庆大霉素、白介素2（interleukin-2,IL-2）和重组人胰岛素，重点验证检测方法的专属性、准确度、可重现性、精密度、定量限、线性、范围、耐用性等7个指标。结果:商业化ELISA试剂盒不受TIL注射液溶剂和上清液的干扰。使用各自标准品检测上述3种辅料的回收量与实际加标量的差异在30%以内，而且复孔读数的变异系数均<10%。适用线性检测范围分别为0.3～6.0 ng·mL^-1、31.3～2000.0 pg·mL^-1、3.0～160.0 mIUg·L^-1。待测TIL样本中庆大霉素、IL-2和重组人胰岛素残留浓度最高值分别为<0.3 ng·mL^-1、37.4 pg·mL^-1和<0.1 mIU·L^-1,均符合对应的放行标准。结论:现有ELISA试剂盒在检测3种培养基辅料残留时都具有良好的灵敏度、准确性和稳定性，可以快速定量分析TIL产品中是否含有潜在的杂质成分。更多还原

【Abstract】 Objective:To establish methods that can efficiently and reliably quantify several residual substances in expanded tumor-infiltrating lymphocytes（TIL） before clinical applications. Methods:Quantitative enzyme-linked immunosorbent assays（ELISA） were used to determine the concentrations of gentamicin, interleukin-2（IL-2） and recombinant human insulin in TIL products, with a focus on specificity, accuracy, reproducibility, precision, linearity, as well as the limit, range and durability of detection. Results:In testing multiple batches of TIL products, neither TIL suspension buffer nor TIL supernatant interfere with the performance of commercial ELISA kits, and the yields of spiked analytes from test samples closely matched the known amounts（<30% deviations in either direction）. The optimal and linear range of detection for gentamicin, IL-2 and recombinant human insulin were 0.3-6.0 ng·mL^-1, 31.3-2000.0 pg·mL^-1, and 3.0-160.0 mIU·L^-1, respectively. The highest concentrations for residual analytes detected in TIL products were <0.3 ng·mL^-1, 37.4 pg·mL^-1, and <0.1 mIU·L^-1, respectively. These parameters all met quality standards for clinical usage. Conclusion:Existing ELISA kits are sensitive, accurate and reliable in measuring three kinds of the above residues in expanded TIL and can serve as rapid tools in quality control procedures.更多还原