摘要
目的建立免疫小鼠体内抗人丙氨酸氨基转移酶(ALT)抗体的间接ELISA检测方法,以期作为融合后杂交瘤细胞株的筛选体系。方法利用人ALT为包被抗原,以1∶40 000稀释的辣根过氧化物酶标记的羊抗鼠IgG为二抗,采用棋盘滴定法确定抗原和对照血清(阴性、阳性)最佳工作浓度及ELISA封闭液的浓度建立ELISA体系。结果包被抗原和血清的适宜浓度或稀释度分别为1∶5 000和1∶1 000;用4%BSA封闭18 h^24 h效果最好。结论建立的ELISA方法稳定,简便易行,能准确检测免疫小鼠血清抗体的效价,并可用于抗ALT单克隆抗体杂交瘤细胞株的筛选。
Objective To develop an indirect method of enzyme-linked immunosorbent assay （ELISA） for detecting anti- ALT monoclonal antibodies. This method can be used to screen hybridoma cell lines after fusion. Methods An indirect ELISA system was established with human ALT-coated and HRP-labeled goat anti-mice IgG diluted to 1 ： 40 000. Optimal working concentrations of antigens and reference sera （positive and negative sera） were determined using checker- board titration and optimal concentrations of the blocking solution were determined. Results Appropriate concentra- tions of antigens and reference sera were 1 ： 5 000 and 1 ： 1 000, respectively. Four percent BSA blocking for 18 h --24 h was most effective. Conclusion This method is simple and reliable. It can accurately examine the titer of immunized mice and screen hybridoma cell lines for anti-ALT monoclonal antibodies.
出处
《中国病原生物学杂志》
CSCD
2010年第2期127-129,共3页
Journal of Pathogen Biology
