单抗体夹心ELISA定量检测人神经生长因子试剂盒的研制  

Development of kits of a single antibody sandwich ELISA for quantitative detection of human nerve growth factor

作　　者：徐春娥[1] 王威[2] 丁锐[2] 纪宏[2] 吴小红[3] 田光[3] 

机构地区：[1]北京市药品审评中心,北京100061 [2]北京市药品检验所,北京100053 [3]军事医学科学院微生物流行病研究所,北京100071

出　　处：《中国当代医药》2016年第15期8-12,共5页China Modern Medicine

摘　　要：目的制备抗人神经生长因子（h NGF）单克隆抗体,并开发一种检测h NGF含量的ELISA试剂盒。方法以重组人神经生长因子抗原与弗氏完全佐剂混合免疫BALB/c小鼠,并采用杂交瘤技术制备抗h NGF的单克隆抗体,然后建立快速检测h NGF含量的ELISA检测试剂盒。结果获得1株可分泌抗h NGF单克隆抗体的杂交瘤细胞株。建立1种检测h NGF含量的ELISA试剂盒,该检测方法的回收率为85%-105%,灵敏度为0.63 ng/ml,变异系数〈10%,且特异性良好。结论成功制备了抗h NGF的单克隆抗体,并建立了一种可快速检测h NGF含量的ELISA试剂盒。Objective To prepare mouse monoclonal antibody against human nerve growth factor（h NGF） and develop an ELISA kits for quantitative detection of h NGF. Methods BALB/c mice were immunized with recombinant human nerve growth factor antigen and Freund′s complete adjuvant,the monoclonal antibody against h NGF was prepared by the technique of hybridoma,and then the ELISA detection kit for rapid detection of h NGF content was established. Results1 hybridoma cell line secreting monoclonal antibodies against h NGF was obtained.1 ELISA kits for the detection of h NGF content was established.The recovery rate of the detection method was 85%-105%,the sensitivity was 0.63 ng/ml,the coefficient of variation was less than 10%,and the peculiarity was good. Conclusion A monoclonal antibody against h NGF is successfully prepared,a rapid and precise ELISA kit is established for detection of rh NGF content.

关 键 词：人神经生长因子 单克隆抗体 ELISA定量检测 

分 类 号：R-331[医药卫生]