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黄曲霉毒素B1(AFB1)与羧甲基羟胺半盐酸盐(CMO)反应,在其分子上引入羧基,生成黄曲霉毒素B1肟化物(AFB1-O),对其肟化物进行薄层层析,紫外扫描和质谱鉴定,并测定肟化率;采用碳二亚胺盐酸盐(EDC)和二环已基碳二亚胺(DCC)的方法来合成黄曲霉毒素B1免疫抗原和包被原,并对两种抗原进行鉴定,计算偶联比和抗原蛋白质浓度。结果表明:AFB1-O合成成功,其肟化率为85.7%,AFB1-O与牛血清白蛋白(BSA)及鸡卵清白蛋白(OVA)偶联成功,偶联比分别为5.4∶1和4.8∶1,为将来制备AFB1单克隆抗体提供了参考。
Aflatoxin B1 (AFB1) reacts with carboxymethylhydroxylamine hemihydrochloride (CMO) to introduce carboxyl groups on its molecule to produce aflatoxin B1 oxime (AFB1-O), which is subjected to thin layer chromatography , UV scanning and mass spectrometry identification, and determination of the oximation rate. The aflatoxin B1 immunogen and the original coating were synthesized using carbodiimide hydrochloride (EDC) and dicyclohexylcarbodiimide (DCC) , And two antigens were identified to calculate the coupling ratio and antigen protein concentration. The results showed that the synthesis of AFB1-O was successful with the oximation rate of 85.7% and the coupling of AFB1-O with bovine serum albumin (BSA) and chicken egg albumin (OVA) successfully. The coupling ratios were 5.4:1 and 4.8 respectively : 1, provide a reference for the future preparation of AFB1 monoclonal antibody.