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目的:制备抗3,4-亚甲基二氧甲基苯丙胺(MDMA)小鼠单克隆抗体(mAb),初步鉴定其特性,为MDMA快速检测方法的开发奠定基础。方法:将摇头丸与丁二酸酐反应生成摇头丸丁二酸酐的羧基衍生物(MDMA-HS),通过活性酯法MDMA-HS偶联于载体牛血清白蛋白(BSA)和卵清白蛋白(OVA),合成免疫抗原MDMA-HS-BSA和检测抗原MD-MA-HS-OVA。将MDMA-HS-BSA免疫BALB/c小鼠,通过杂交瘤技术获得杂交瘤细胞株19A11。结果:将该细胞株注射BALB/c小鼠制备腹水mAb。腹水纯化后效价为1∶1.6×105,属于IgG1型。MDMAmAb的灵敏度为0.93μg/L,IC50为6.07μg/L。mAb与大麻、麻黄碱、氯胺酮、苯丙胺、甲基苯丙胺、吗啡、可卡因的交叉反应率均小于0.3%。结论:制备出的MDMAmAb为研制MDMA的快速检测方法奠定了基础。
OBJECTIVE: To prepare monoclonal antibody (mAb) against 3,4-methylenedioxymethamphetamine (MDMA) mouse and to identify its characteristics initially, which laid the foundation for the development of MDMA rapid detection method. Methods: MDMA-HS was prepared by reacting MDMA with succinic anhydride. MDMA-HS was coupled to carrier bovine serum albumin (BSA) and ovalbumin (OVA) by active ester method. The synthetic immunogens MDMA-HS-BSA and the detection antigen MD-MA-HS-OVA. BALB / c mice were immunized with MDMA-HS-BSA and hybridoma cell line 19A11 was obtained by the hybridoma technique. Results: BALB / c mice were injected with this cell line to prepare ascites mAb. Ascites purified titers of 1: 1.6 × 105, belonging to IgG1 type. The sensitivity of MDMA mAb was 0.93 μg / L and the IC50 was 6.07 μg / L. The cross-reaction rates of mAb with marijuana, ephedrine, ketamine, amphetamine, methamphetamine, morphine and cocaine were all less than 0.3%. Conclusion: The prepared MDMA mAb lays the foundation for the rapid detection of MDMA.