白三烯C4的酶免疫测定法

Enzyme immunometric assay for leukotriene C4.

作者信息

Volland H, Vulliez Le Normand B, Mamas S, Grassi J, Créminon C, Ezan E, Pradelles P

机构信息

SPI-BIO, Commissariat à l'Energie Atomique, Gif sur Yvette, France.

出版信息

J Immunol Methods. 1994 Sep 30;175(1):97-105. doi: 10.1016/0022-1759(94)90335-2.

DOI:10.1016/0022-1759(94)90335-2

PMID:7930643

Abstract

An enzyme immunometric assay of LTC4 named SPIE-IA is described. The assay involves different sequential steps: (1) immunocapture of LTC4 by monoclonal anti-LTC4 antibodies coated on 96-well microtiter plates; (2) cross-linking of LTC4 via its amino group to the wells using glutaraldehyde; (3) treatment with HCl; (4) measurement of linked LTC4 using the same monoclonal anti-LTC4 antibodies labeled with acetylcholinesterase. A minimal detectable concentration of 2 pg/ml after 60 min of enzymatic reaction was obtained. Cross-reactivity was less than 15% with LTD4 or LTE4. The coefficient of variation was less than 6% in the 20-1000 pg/ml range. Good correlation was observed between SPIE-IA and a competitive enzyme immunoassay for biological samples. The different sequential steps of the assay are investigated.

摘要

描述了一种名为SPIE-IA的白三烯C4（LTC4）酶免疫测定法。该测定法包括不同的连续步骤：（1）用包被在96孔微量滴定板上的抗LTC4单克隆抗体免疫捕获LTC4；（2）使用戊二醛通过其氨基将LTC4交联到孔中；（3）用盐酸处理；（4）使用标记有乙酰胆碱酯酶的相同抗LTC4单克隆抗体测量交联的LTC4。酶促反应60分钟后获得的最低可检测浓度为2 pg/ml。与白三烯D4（LTD4）或白三烯E4（LTE4）的交叉反应率小于15%。在20-1000 pg/ml范围内变异系数小于6%。观察到SPIE-IA与生物样品的竞争性酶免疫测定法之间具有良好的相关性。对该测定法的不同连续步骤进行了研究。