酶联免疫法快速测定原料乳中κ-酪蛋白含量

The determination of bovineκ-casein by enzyme-linked immunosorbent assays

用牛κ-酪蛋白纯品免疫新西兰白兔,制得兔抗牛κ-酪蛋白的多克隆抗体。以牛κ-酪蛋白包被抗原,自制抗体为一抗,辣根过氧化物酶标记的羊抗兔IgG为二抗,邻苯二胺(OPD)为底物,建立了牛乳中κ-酪蛋白的间接ELISA检测方法。检测的线性范围是25～1400ng/mL,相关系数是0．9831,回收率在95．83％～125．83％之间,证明可以用酶联免疫法简单快速地测定牛乳中的κ-酪蛋白,因为该蛋白在酪蛋白中含量稳定,所以可以计算出牛乳中酪蛋白的含量。显然该方法可以用于牛乳掺杂使假的鉴别和原料奶酪蛋白含量是否合格的快速检测。

The polyclonal antibody was prepared by immunizing New Zealand rabbits with bovine k- casein. Enzyme-Linked Immunosorbent Assays （ELISA） was developed for bovine k-casein. The standard curve was linear between 25 and 1400ng/mL with regression coefficients of 0.9831. The recovery rates were between 95.83% - 125.83%. It is proved that using ELISA can determinate the content of bovine k-casein easily and effectively. Because the content of k-casein is so steady that its content can be calculated easily. Obviously, this method can be used to find out the adulteration of raw milk and quantify the content of bovine k-casein.