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目的人工合成黄芩苷(BAL)的完全抗原并进行鉴定。方法采用碳二亚胺两步法,将BAL分别与牛血清白蛋白(BSA)和多聚赖氨酸(PLL)相偶联。用紫外扫描的方法测定其偶联比,用动物免疫的方法测定其抗原性。结果 BAL-BSA的偶联比为6∶1,BAL-PLL的偶联比为7∶1,间接ELISA实验结果表明,6只BAL人工抗原免疫的小鼠均有特异性抗体产生,抗体效价为1∶3 000以上。间接竞争ELISA实验证实BAL可特异性地阻断上述反应的发生。结论该方法成功合成了BAL的人工抗原,并且在免疫小鼠的血清中产生了针对BAL的抗体。
Objective To synthesize and identify the complete antigen of baicalin (BAL). Methods BAL was coupled with bovine serum albumin (BSA) and polylysine (PLL) respectively by carbodiimide two-step method. The coupling ratio was determined by UV scanning method, and its antigenicity was measured by animal immunization method. Results The conjugation ratio of BAL-BSA was 6:1 and the coupling ratio of BAL-PLL was 7:1. The results of indirect ELISA showed that all the 6 BAL antigens were immunized with specific antibodies. The antibody titers 1: 3000 or more. Indirect competitive ELISA test confirmed that BAL can specifically block the occurrence of the above reaction. Conclusion This method successfully synthesized BAL artificial antigen and produced antibodies to BAL in the serum of immunized mice.