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目的人工合成紫铆素(butin)的完全抗原并进行鉴定。方法采用氯乙酸钠(SMCA)与紫铆素中的羟基反应形成羧甲基醚衍生物(紫铆素-SMCA),再通过羧基与载体蛋白(BSA、OVA)偶联形成完全抗原,以TLC法鉴别是否偶联产生新的大分子化合物,并以紫外扫描分析完全抗原的偶联比率,将得到的免疫原紫铆素-BSA按免疫程序免疫Balb/c小鼠。结果 TLC显示已经形成新的大分子化合物;紫外扫描法测定紫铆素-BSA的偶联比为6∶1,紫铆素-OVA的偶联比为5∶1。采用间接酶联免疫测定,产生的抗血清的稀释度可以达到1∶4 000。结论经过TLC法、紫外扫描法和免疫方法鉴定,成功合成了紫铆素人工抗原。
Objective To synthesize and identify the complete antigen of butin. Methods Sodium carboxymethyl sulfate (SMCA) was reacted with hydroxyl group in pridin to form carboxymethyl ether derivative (VBR-SMCC). Then, the complete antigen was formed by the coupling of carboxyl group and carrier protein (BSA, OVA) The method was used to identify whether the new macromolecular compound was coupled and the ratio of the complete antigen was analyzed by UV scanning. The immunogenic pridin-BSA was immunized to Balb / c mice by immunization. Results TLC showed that a new macromolecular compound had been formed. The coupling ratio of pridin-BSA was 6:1 as determined by UV-scan, and the coupling ratio of pridin-OVA was 5:1. Using indirect enzyme-linked immunoassay, the resulting antisera can be diluted to 1: 4000. Conclusion The method of TLC, UV-scanning and immunological methods were successfully used to synthesize rivastigmine artificial antigen.