乙草胺人工抗原的合成及其单克隆抗体的制备

Synthesis of Artificial Antigen and Preparation of Monoclonal Antibody for Acetochlor

摘要: 通过乙草胺与3-巯基丙酸在碱性条件下反应合成了半抗原——乙草胺-巯基丙酸(AMPA)。采用活性酯法将半抗原AMPA分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联制备了人工抗原。通过紫外光谱测定了AMPA与BSA及OVA间的分子结合比分别为41和26。用AMPA-BSA免疫小鼠,制备得到的单克隆抗体的效价为1∶ 16×104。以AMPA-OVA作包被抗原,用乙草胺单克隆抗体建立了ELISA检测方法,IC50值为0.55 μg/L,方法的检测范围为0.04~5 μg/L,检测限为0.04 μg/L。

Abstract: The hapten of acetochlor-MPA(AMPA) was synthesized by the reaction of acetochlor with 3-mercaptopropionic acid in alkaline solution.The artificial antigens (AMAP-BSA/OVA) were synthesized by coupling of molecules between hapten and BSA,OVA respectively with active ester method.The molar ratio was 41,26 for AMPA-BSA and AMPA-OVA respectively detected by UV spectro-photometer.Mice were immunized by injection of antigen AMPA-BSA.The monoclonal antibody was produced with titer of 1∶ 16×104.The ELISA method based upon AMPA-OVA and monoclonal antibody was established.This ELISA method showed its IC50 value was 0.55 μg/ L,the response range for acetochlor was 0.04~5 μg/L,and limit of detection was 0.04 μg/L.