糖化牛血清白蛋白及其抗体的制备

目的制备特异性强、效价高的抗晚期糖化终末产物(AGEs)的多克隆抗体,以用于AGEs免疫化学方法的检测。方法用牛血清白蛋白与葡萄糖在体外共同孵育,制备糖化牛血清白蛋白(AGEs-BSA),经凝胶层析纯化后,免疫新西兰家兔,获取抗AGEs-BSA抗血清,经硫酸铵盐沉淀法制备多克隆抗体。结果荧光光谱和十二烷基硫酸钠聚丙烯酰胺凝胶电泳鉴定结果表明,成功制备出AGEs-BSA。ELISA结果证实,制备出特异性强、效价高的抗AGEs的多克隆抗体,其效价高达1:2 000。结论应用高糖孵育BSA可以制备出AGEs-BSA,通过AGEs-BSA免疫新西兰家兔成功制备特异性强、效价高的抗AGEs的多克隆抗体,可以用于AGEs免疫化学方法的检测,并为进一步研究AGEs在阿尔茨海默病发病中的作用奠定基础。

Preparation of AGEs-BSA and AGEs-BSA antibody

Objective To obtain high titer and specific polyclonal antibody of advanced glycation end products(AGEs) for immunochemical detection of AGEs.Methods BSA and glucose were incubated together in vitro to prepare AGEs-BSA,purifie it through gel chromatography,immunize New Zealand white rabbits to obtain the anti-AGEs serum,and prepare the polyclonal antibody of AGEs through the sedimentation of saturate ammonium sulfate.Results The purified AGEs-BSA possess special fluorescence spectrum and showed smear shape in SDS-PAGE.AGEs-BSA antibody has strong specificity through the detection of ELISA.The titer of antibody by the detection of ELISA is 1:2000.Conclusion AGEs-BSA can be successfully prepared through the incubation of glucose and BSA.The high titer of polyclonal antibody of AGEs can be successfully prepared by using AGEs-BSA to immunize New Zealand white rabbits and be used to detect AGEs by immunochemical method.These results will help to further study the function of AGEs in the process of Alzheimer′s disease.