小鼠肿瘤坏死因子-α光激化学发光免疫法的建立

Establishment of Light Initiated Chemiluminescence Assay of Mouse TNF-α

采用光激化学发光免疫法（LICA）建立快速定量检测小鼠mTNF-α。用两株mTNF-α不同表位的单克隆抗体（McAb）,一株McAb包被发光微粒,另一株为生物素化McAb,两者与链亲和素包被的感光微粒构建双McAb夹心法mT-NF-α-LICA。数据采用双对数函数处理程序。结果表明：方法灵敏度为0.5ng/L;ED20、ED50、ED80分别为14.96 ng/L、77.5 ng/L、401.0 ng/L;可测范围为0.5~1200 ng/L;批内和批间CV分别为7.8%和10.5%;平均回收率为100.3%。mTNF-α-LICA与ELISA有较好的相关性。本文建立的mTNF-α-LICA法是mTNF-α检测中灵敏的方法,该方法稳定性好,具有很好的应用前景。

To establish a mouse TNF-α level, two quantitative light initiated chemiluminescence assay （LICA） for the determination of specific McAbs were selected. One was coated on iuminescence particles and the other was conjugated with biotin. These two McAb with sensitizer particles which were coated with streptavidin were used to constructed mTNF-α-LICA system. The results showed that the analytical sensitivity was 0.5ng/L. The 20% , 50% and 80% inhibition binding effect dose were 14.96 ng/L, 77.5 ng/L and 401.0ng/L,respectively. The detection range was from 0.5 ng/L to 1200 ng/L. The intra- and inter- assay CV of mTNF-α-LICA were 7.8% and 10.5% ,respectively, and the mean recovery rate was 100.3%. The results detected with mT- NF-et-LICA were well correlated with ELISA. The mTNF-α-LICA established in this study was a sensitive, sim- ple and convenient method in detecting mTNF-α level. The mTNF-α-LICA with high sensitivity and wide optimal range showed a great application prospect.