摘要
目的通过人工抗原的构建 ,制备针对槲皮素的特异性抗体。方法采用混合酸酐法合成的槲皮素卵清蛋白结合物作为包被抗原 ,将槲皮素与载体蛋白牛血清白蛋白 (BSA)结合 ,制成人工抗原。用人工抗原免疫家兔 ,制备了针对槲皮素的特异性抗体 ,并用双向琼脂扩散试验和 EL ISA方法对抗体进行了鉴定。IR、UV、TL C和熔点分析表明 ,中间体结合物为新的化合物。结果理想的包被抗原浓度为 40 0 ng/ ml,相应抗血清工作浓度为 1:16 0 0。结论运用混合酸酐法可以合成槲皮素人工抗原 。
ObjectiveTo build the artificial immunogen and product specific antibody.MethodsThe hapten Quercetin was coupled to BSA and OVA with mixed anhydride method and the Quercetin carrier protein conjugations were synthesized. Quercetin HS OVA was used as coating antigen. The antibody was obtained successfully by immunizing big ear rabbits with Quercetin HS BSA,the polyclonal anti quercetin antibodies were developed,which were determined with double agar diffusion test and ELISA. IR,UV,TLC and m.p.for qualitative analysis were also performed. It can be confirmed that the middle conjugations were produced. Results It was shown that the ideal coating antigen concentration was 400 ng/ml,the available antibody dilution was 1:1 600 ConclusionThe artificial immunogen could be synthesized by mixed anhydride method and the specificity of the polyclonal anti quercetin antibodies could be produced.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2000年第5期383-386,共4页
Immunological Journal
基金
湖北省重点资助科技基金项目!(98190802)
关键词
槲皮素
抗体
混合酸酐法
ELISA
BSA
quercetin
antibodies
double agar diffusion test
enzyme- linked immunosorbent assay
