蛋清中食物不耐受组分分析

Analysis the protein components which caused egg intolerance

目的 分析引起患者鸡蛋不耐受的主要蛋白组分.方法 用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate polyacrylamide gel electropheresis,SDS-PAGE）分析蛋清粗提取物中的蛋白组分.以鸡蛋不耐受患者血清特异性IgG作为识别抗体,经Western blotting和间接酶联免疫吸附试验（enzyme linked immunosorbent assay,ELISA）分析与患者血清特异性IgG结合的蛋白成分.结果 蛋清粗提物SDS-PAGE电泳结果主要有4个条带,相对分子质量分别为：14×10^3、34×10^3、45×10^3和76×10^3,根据相对分子质量推测分别为溶菌酶、卵类黏蛋白、卵清白蛋白和卵转铁蛋白.患者血清的免疫印迹结果主要呈现4个条带,分别对应溶菌酶、卵类黏蛋白、卵清白蛋白和卵转铁蛋白,间接ELISA法结果显示四种纯化抗原均与患者血清有反应.结论 引起患者鸡蛋不耐受的蛋白主要有两种,分别为卵类黏蛋白和卵清白蛋白,其相对分子质量分别为34×10^3和45×10^3.

Objective To analysis the protein components of egg leading to egg intolerance. Methods The components of protein extracted from egg were analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE）. The serum of patients with egg intolerance as the identification antibody. Westeru blotting and indirect enzyme linked immunosorbent assay （ELISA） were used to identify the major components of egg protein which bound with the egg intolerance patients＇ serum specific IgG. Results The SDS- PAGE results showed four straps which relative molecular weight were 14×10^3, 34×10^3, 45×10^3 and 76×10^3. They were lysozyme, ovomueoid, ovalbumin and ovotransferrin, respectively according to the relative molecular mass. The western blotting showed four straps which corresponding with lysozyme, ovomucoid, ovalbumin and ovotransferrin. The results of indirect ELISA method showed that the four kinds of purified proteins all had reaction with patients＇ serum. Conclusion There are two main proteins including ovomucoid and ovalbumin caused egg intolerance. The relative molecular weight are 34×10^3 and 45×10^3, respectively.