摘要
目的:建立高灵敏度和特异性的腺激肽释放酶2(hk2)酶联免疫吸附试验(ELISA)。方法:以hk2单克隆抗体(hk2mAb)为包被抗体,hk2多克隆抗体(hk2pAb)和辣根过氧化酶(HRP)标记抗IgG为检测抗体,建立检测hk2双抗体夹心ELISA法,并对其灵敏度、精密度、准确度和稳定性等方法学进行评价。结果:hk2mAb、hk2pAb和HRP标记IgG的最佳稀释工作浓度分别为1∶100、1∶2 000和1∶8 000。该法线性范围为50～800 ng/L、线性良好(R2=0.99)、灵敏度为28 ng/L、批内和批间CV分别为5.78%和7.56%、平均回收率为99.4%。结论:双抗体夹心ELISA法检测人血清hk2,其敏感性高、特异性强、准确度好,可满足人血清和组织细胞培养液中hk2水平的检测。
Objective：To develop a double antibody sandwich ELISA method for detecting human glandular kallikrein 2 （hk2 ）. Methods ：The monoelonal, polyclonal antibody and a HRP-labeled conjugate of another anti-IgG to hk2 were prepared by ourselves, and a double-antibody-sandwich ELISA method for the detection of serum hk2 was established, then initially study the research methodology of detection. Results ：The best working concentrationof hk2 mAb ,hk2 pAb and HRP-anti-IgG were 1： 100,1：2 000 and 1：8 000, The linear detection was 50 to 800 ng/L（ R^2 =0.99）. The sensitivity was 28 ng/L, and the mean recoveries of samples was 99.4%. while the intra- and inter-assay precisions were 5.78% and 7.56%. Conclusion： Double-antibody sandwich ELISA assay of serum hk2 has good sensitivity, specificity, accuracy, and it can be used to measure hk2 level in human plasma or serum and the supernatant of cell culture.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2013年第4期416-418,共3页
Chinese Journal of Immunology
基金
浙江省教育厅科研基金资助项目(Y201120586)
作者简介
谢克俭（1966年-），男，高级实验师，主要从事分子生物学和临床免疫学教学和科研工作，E-mail：xkj@wzmc．edu．net。
