人血清中抗鼠抗体间接ELISA检测方法的建立及临床应用  

Development and Clinical Application of Indirect ELISA for Human Anti-Mouse Antibody in Human Serum

作　　者：郭长福[1] 张囡[1] 李茜[1] 黎维勇[1] 孙可芳[1] 胡豫[2] 张爱华[1] 

机构地区：[1]武汉生物制品研究所免疫学研究室,武汉430060 [2]华中科技大学同济医学院附属协和医院,武汉430060

出　　处：《中国生物制品学杂志》2009年第7期716-718,共3页Chinese Journal of Biologicals

摘　　要：目的建立检测人血清中抗鼠抗体(HAMA)的间接ELISA法,并进行临床标本检测。方法采用间接ELISA法,选择最佳实验条件,确定阴阳性血清判定的界值,并进行方法学验证;用该方法检测WuTacⅠ期临床观察的健康志愿者血清抗鼠抗体。结果间接ELISA的最佳试验条件为:抗原包被浓度0.313μg/ml,血清稀释度1∶160,酶标抗体稀释度1∶80 000;阴阳性血清界值为0.210;该方法检测HAMA阳性标本的试验内和试验间变异系数分别为7.46%和5.48%,检测HAMA阴性标本的试验内和试验间变异系数分别为8.03%和14.7%;与马、山羊、兔、猴等动物血清无交叉反应;健康志愿者HAMA约在用药后10～14 d产生。结论已成功建立了检测人血清中抗鼠抗体的间接ELISA法,符合我国生物药品临床试验检测的要求。Objective To develop an indirect ELISA for human anti-mouse antibody （HAMA） in clinical serum specimens. Methods An indirect ELISA for HAMA in sera was developed, and the experimental condition was optimized, based on which the cutoff values for judgment of positive and negative results were determined, and the developed ELISA was verified methodologically. The serum specimens of healthy volunteers injected with McAb WuTac in clinical trial phase I were determined by the developed ELISA. Results The optimal antigen coating concentration of the developed ELISA was 0. 313 μg/ml, and the optimal dilutions of serum and enzyme-labeled antibody were 1 ： 160 and 1 ： 80 000 respectively. The cutoff A value of positive result was 0. 210. The inter-and intra-CV values of the developed ELISA for determination of HAMA-positive specimens were 7.46% and 5.48%, and those for HAMA-negative specimens were 8. 03% and 14. 7%, respectively. The positive serum specimens confirmed by the developed ELISA showed no cross reaction with horse, goat, rabbit and monkey sera. HAMAs were induced in healthy volunteers 10-14 d after injection with WuTac. Conclusion An indirect ELISA for HAMA in human serum was successfully developed, which met the requirements for clinical trial of biological products in China.

关 键 词：人抗鼠抗体 WuTac 间接ELISA 

分 类 号：R392.7[医药卫生—免疫学] Q819[医药卫生—基础医学]