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

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1：16×10^4。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0.55μg/L，方法的检测范围为O．04～5μg／L，检测限为O．04μg/L。

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 x 10^4. 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.