酪蛋白和乳球蛋白的抗体及酶标物制备研究

Preparation of β-Casein and β-Lacotoglobuli Antibodies and Enzyme-labeled Antigens

选用牛乳中的β-酪蛋白(β-CN)和β-乳球蛋白(β-Lg)为抗原对鸡体免疫,从鸡卵黄中用水稀释法及亲和色谱(HiTrapTM IgY Purification HP)纯化特异性鸡卵黄抗体(IgY)。PAGE电泳分析IgY纯度高达90%,ELISA效价为1:4096。用过碘酸钠氧化法进行抗原的过氧化物酶(HRP)标记,HRP与β-CN和β-Lg的分子个数比分别为1:1和1.5:1的标记效果最好,β-CN-HRP和β-Lg-HRP的效价高达1:640,其ELISA工作浓度分别为1:80和1:40。特异性IgY和酶标抗原为特异检测牛乳蛋白的酶联免疫吸附(ELISA)分析提供必要条件。

Hens were immunized with the antigen of bovine- β-casein （β-CN） and β-lactoglobulin （β-Lg）, and the egg yolk immunoglobulin （IgY） of the hens was purified by the methods of water diluting and affinity chromatography （HiTrap^TM IgY Purification HP）. The purity of IgY is as high as 90% by PAGE and the ELISA titer is 1：4096. In the NaIO4 labeled method, the antigens were conjugated with horseradish peroxidase （HRP）, respectively. The molar ratios between HRP and β-CN or β-Lg for the best labding effect are 1 ;1 and 1.5;1, respectiwly. The β-CN-HRP and β-Lg-HRP titers are both 1：6407 and their operation concentrations arc 1：80 and 1：40, respectively.