二氯喹啉酸人工抗原的合成与鉴定.pdfVIP

中国农业科学 2005,38(1):86-90 Scientia Agricultura Sinica 二氯喹啉酸人工抗原的合成与鉴定 朱国念，毛黎娟，施海燕，黄雅丽，程敬丽 （浙江大学农药与环境毒理研究所，杭州 310029 ） 摘要：以除草剂二氯喹啉酸（quinclorac ，Q ）、氯化亚砜、氨基己酸和氨基丁酸等为起始原料，经2 步化学反 应分别合成了2种二氯喹啉酸半抗原：6-(3,7-二氯-8-喹啉甲酰基)氨基己酸（QC）和4-（3,7-二氯-8-喹啉甲酰 基）氨基丁酸（QB ）。并通过碳化二亚胺法和混合酸酐法将Q、QC和QB分别与载体蛋白（BSA 、OVA ）偶联制备了 二氯喹啉酸的免疫抗原和包被抗原。再将Q-BSA、QC-BSA和QB-BSA分别免疫新西兰大白兔获得了3种抗二氯喹啉酸 的多克隆抗体：抗Q-BSA抗体(Q-Ab)、抗QC-BSA抗体(QC-Ab)和抗QB-BSA抗体(QB-Ab)，当相应的包被抗原浓度 均为 4 µg·ml-1 3 6 6 时，相应的冻干粉效价分别为2×10 ，1.2×10 和 1.6×10 。由于Q-Ab效价偏低，不适合用于检测。采 用同源间接ELISA竞争法测定时，其他2种抗体（QC-Ab 、QB-Ab ）的抑制中浓度(IC50)均大于 50 mg·L-1 ，检测灵 敏度也较低。因此本文对异源间接竞争ELISA法进行了研究，结果表明，异源反应的检测灵敏度高于同源反应， 其中QB-Ab 的IC50为 0.4 mg·L-1 ；最低检测限为 0.006 mg·L-1 。 关键词：二氯喹啉酸; 半抗原; 人工抗原; 合成 Synthesis and Identification of Artificial Antigen Against Quinclorac ZHU Guo-nian, MAO Li-juan, SHI Hai-yan, HUANG Ya-li, CHENG Jing-li (Institute of Pesticide and Environmental Toxicology, Zhejiang University, Hangzhou 310029) Abstract: In this paper, two haptens of quinclorac(Q), 6-[(3,7-dichloro-8-quinolino-formacyl) amino-hexanoic acid (QC) and 4-[(3,7-dichloro-8-quinolino-formamino)] butanoic acid (QB), were synthesized by using quinclorac and amino-hexanoic acid or amino-butanoic acid as original materials. These two haptens and quinclorac were separately conjugated to bovine serum albumin (BSA) with the modified active ester method. Three coating antigens, Q-OVA, QC-OVA and QB-OVA were also synthesized with the mixed-anhydride method. As a result of the study, three types of antibody (Q-Ab, QC-Ab and QB-Ab) were produced successfully. Their titers were 2×103 6 6 -1 ，1.2×10 a