肿瘤浸润淋巴细胞注射液中多种杂质残留成分的定量分析探索  

Quality Control for Expanded Tumor-infiltrating Lymphocytes (TIL):Quantification of Several Residual Substances

作　　者：王涛 周足力[2] 肖蓉心 初明[3,4] 王星星 唐建明 王铁山 陈萌 张秀军 WANG Tao;ZHOU Zuli;XIAO Rongxin;CHU Ming;WANG Xingxing;TANG Jianming;WANG Tieshan;CHEN Meng;ZHANG Xiujun(The Fifth Medical Center of the PLA General Hospital,Beijing 100039,China;People′s Hospital of Peking University,Beijing 100044,China;School of Basic Medical Sciences,Peking University,Beijing 100191,China;Key Laboratory of Medical Immunology,National Health Commission,Beijing 100191,China;Aeonvital Institute of Clinical and Translational Immunology,Beijing 102600,China;Beijing University of Chinese Medicine,Beijing 100029,China;China National Institute of Standardization,Beijing 100191,China)

机构地区：[1]解放军总医院第五医学中心,北京100039 [2]北京大学人民医院,北京100044 [3]北京大学基础医学院,北京100191 [4]国家卫生健康委员会医学免疫学重点实验室,北京100191 [5]北京循生免疫医学转化研究院,北京102600 [6]北京中医药大学,北京100029 [7]中国标准化研究院,北京100191

出　　处：《中国医药导刊》2023年第4期403-409,共7页Chinese Journal of Medicinal Guide

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。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.

关 键 词：肿瘤浸润淋巴细胞 细胞培养基辅料 杂质残留检测 庆大霉素 白介素2 重组人胰岛素 

分 类 号：R73[医药卫生—肿瘤] R730.5[医药卫生—临床医学] R917