黄芩苷完全抗原的合成鉴定及其多克隆抗体的制备

目的: 研究黄芩苷作为一种半抗原分子,通过与载体蛋白偶联形成完全抗原后,使免疫动物血清中产生特异性抗体的可能性,为进一步建立黄芩苷的免疫分析测定方法提供实验依据。 方法: 采用活性酯法将黄芩苷(BAL)与牛血清白蛋白(BSA)偶联,制备黄芩苷完全抗原(BAL-BSA),并采用红外吸收光谱法、SDS-PAGE法、MALDI-TOF-MS法对其进行鉴定。用此完全抗原免疫动物制备抗血清,通过间接ELISA法检测其抗体效价。 结果: 完全抗原BAL-BSA中BAL与BSA的偶联比为3: 1;用此完全抗原免疫的家兔产生针对BAL的多克隆抗体,抗体效价最高可达1: 8000。 结论: 成功合成了BAL的完全抗原BAL-BSA,且该抗原具有良好的反应原性与免疫原性。

Objective: To synthesize baicalin(BAL) complete antigen through coupling BAL which could be taken as a hapten to bovine serum albumin(BSA),to investigate the possibility of this complete antigen as an immunogen in inducing specific antibody from sera of animals immunized with it,and provide some experimental data for establishing the immunoassay for BAL. Methods: The complete antigen(BAL-BSA) was prepared by coupled BAL to BSA using activated ester method,and identified by infrared absorption spectrometry,SDS-PAGE and MALDI-TOF-MS.Rabbits were immunized with BAL-BSA to produce antiserum against BAL.The antisera titer were detected by indirect ELISA. Results: The coupling ratio of BAL to BSA was 3: 1.The indirect ELISA showed that animals immunized with BAL-BSA produced specific antibody which reacted specifically to BAL with the titer up to 1: 8000. Conclusion: The successfully synthesized complete antigen(BAL-BSA) has good immunogenicity and reactogenicity.