格列本脲人工抗原的合成与鉴定

目的:合成格列本脲人工抗原。方法:采用对氨基苯甲酸法制备半抗原,将半抗原分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)通过碳二亚胺法偶联制备免疫原(Gli-BSA)和包被原(Gli-OVA),利用紫外扫描进行抗原的化学鉴定,通过免疫原免疫Balb/c小鼠,间接酶联免疫吸附法测定抗血清进行生物鉴定。结果:制备了Gli-BSA、Gli-OVA的人工抗原,经紫外光谱扫描,偶联比分别为4:1和17.7:1。免疫小鼠后获得抗血清的效价均达到32000以上,半抑制质量浓度为10μg/mL。结论:成功合成了格列本脲人工抗原,并获得了格列本脲抗体,为格列本脲的免疫学检测方法进一步研究提供了实验基础。

Synthesis and Identification of Artificial Antigen for Glibenclamide

Objective: To synthesize artificial antigen for glibenclamide.Methods: Hapten was synthesized by diazo reaction with p-aminobenzoic acid,and conjugated separately with BSA(bovine serum albumin) and OVA(ovalbumin) by carbodiimide method.Immunogen and coating antigen were obtained and designated as Gli-BSA and Gli-OVA,respectively.The synthesized artificial antigens were identified by mass spectrometry and UV spectroscopy.The immunogen was used to immunize Balb/c mice and the generated anti-serum was analyzed by indirect ELISA.Results: Gli-BSA and Gli-OVA were successfully synthesized with a coupling ratio of 4:1 and 17.7:1,respectively.The anti-serum titers after the first,second and third immunization of Balb/c mice were all above 32000 and the IC50 was 10 μg/mL.Conclusion: Artificial antigens for glibenclamide has been successfully synthesized and anti-glibenclamide antibodies have been acquired.This study can provide a basis for further studies on immunological detection of glibenclamide.