Kinghorn N M, Norris C S, Paterson G R, Otter D E
Food Science Section, New Zealand Dairy Research Institute, Palmerston North.
J Chromatogr A. 1995 May 12;700(1-2):111-23. doi: 10.1016/0021-9673(95)00117-6.
The separation of the four major whey proteins by sodium dodecyl sulphate (SDS)-capillary gel electrophoresis (CGE) is described. Whilst commercially purified whey proteins could be analysed using the recommended protocol, the more complex nature of an acid whey and a reconstituted whey protein concentrate (WPC) powder necessitated considerable refinement of the CGE sample buffer. Individual whey proteins in the acid whey and WPC samples were then also separated and quantitated using capillary zone electrophoresis, polyacrylamide gel electrophoresis (PAGE) and HPLC methods and the results were compared. The values obtained for alpha-lactalbumin (alpha-Lac) and beta-lactoglobulin (beta-Lg) were consistent throughout the various methods, although size-exclusion HPLC, SDS-PAGE and SDS-CGE could not separate the two beta-Lg variants or the glycosylated form of alpha-Lac from the beta-Lg. There was considerable variation in the values for the bovine serum albumin and immunoglobulin determined by the different methods and it was concluded that none of the methods could satisfactorily quantitate all four whey proteins.
本文描述了通过十二烷基硫酸钠(SDS)-毛细管凝胶电泳(CGE)分离四种主要乳清蛋白的方法。虽然使用推荐的方案可以分析商业纯化的乳清蛋白,但酸性乳清和重组乳清蛋白浓缩物(WPC)粉末的性质更为复杂,因此需要对CGE样品缓冲液进行大量改进。然后,还使用毛细管区带电泳、聚丙烯酰胺凝胶电泳(PAGE)和HPLC方法对酸性乳清和WPC样品中的单个乳清蛋白进行了分离和定量,并对结果进行了比较。尽管尺寸排阻HPLC、SDS-PAGE和SDS-CGE无法分离两种β-乳球蛋白变体或α-乳白蛋白的糖基化形式与β-乳球蛋白,但通过各种方法获得的α-乳白蛋白(α-Lac)和β-乳球蛋白(β-Lg)的值在整个过程中是一致的。不同方法测定的牛血清白蛋白和免疫球蛋白的值存在相当大的差异,得出的结论是,没有一种方法能够令人满意地定量所有四种乳清蛋白。