鲤鱼小清蛋白的纯化及其过敏原性鉴定

Purification and Allergenicity Identification of Carp Parvalbumin

目的:分离纯化鲤鱼小清蛋白(parvalbumin),并对其进行过敏原性鉴定,为建立鱼类过敏原检测技术奠定基础。方法:采用硫酸铵分级盐析和阴离子交换层析纯化鲤鱼小清蛋白,采用点杂交和特异性IgE检测试剂盒筛选鱼类过敏者血清,应用聚丙烯酰胺凝胶电泳和免疫印迹技术分析确定纯化目标蛋白的性质。结果:硫酸铵分级盐析和阴离子交换层析纯化方法可以得到电泳纯单一目标蛋白;小鼠抗蛙小清蛋白单克隆抗体免疫印迹实验表明,所得纯化蛋白是小清蛋白。此外,免疫杂交结果显示,鱼类过敏者血清能与纯化的小清蛋白发生特异性结合,从而证实了鲤鱼小清蛋白的过敏原性。

Objective:In order to purify and identify the major allergen of carp parvalbumin and develop detection technology for fish allergen.Method:Carp parvalbumin was purified by heat treatment,(NH4)2SO4 fractionation and DEAE-Sepharose chromatography.Dot-blotting and specific IgE screening kit were used to screen the fish allergenic patients serum.SDS-PAGE and Western-blotting were used to further identify and analyze the characteristic of carp parvalbumin.Results:Western-blotting using mouse monoclonal antibody...