脱落酸人工抗原合成及多克隆抗体制备

目的:为了对植物细胞中的脱落酸(ABA)进行定量和定位分析,研究了脱落酸人工抗原的合成以及多克隆抗体的制备。方法:用重氮化法将ABA分别与牛血清白蛋白(BSA)、卵清白蛋白(OVA)联结,得到ABA的免疫抗原和包被抗原,并采用紫外全波长扫描和SDS-PAGE对合成的抗原进行了鉴定。以经过鉴定的抗原免疫白兔,制备出ABA的多克隆抗体;采用间接酶联免疫法(ELISA)对抗血清进行效价检测,通过离子交换层析法获得纯化的抗体。结果:ABA与BSA的平均偶联比为5.3∶1,抗血清效价为1∶16000。结论:人工抗原和多克隆抗体制备成功,为采用ELISA和免疫胶体金技术(ICG)检测ABA提供了有效工具。

Synthesis of Antigen and Preparation of Multi-Clonal Antibody Against Abscisic Acid

Objective:In order to research on the localization and quantization of abscisic acid(ABA)in plant cells,synthesis of antigen and preparation of multi-clonal antibody against ABA were studied.Methods:ABA was conjugated to bovine serum album(BSA)and ovalbumin(OVA)via diazo coupling.The hapten was identified by UV spectrometric scanning and SDS-PAGE.Rabbits were immunized with antigen to obtain the multi-clonal antibody against ABA.Antibody was identified by enzyme-lined immunosorbent assay(ELISA),and purified by ion exchange chromatography.Results:Based on the experiments and calculation,the conjugated ratio of ABA and BSA in ABA-C4'-BSA was 5.3:1,and the titer of the antisera was 1:16 000.Conclusion:The results showed that the synthesis of antigen and preparation of multi-clonal antibody against abscisic acid were successful,which supplied a good tool for the detection of ABA with ELISA and immune colloidal gold(ICG)methods.