Brignon G, Ribadeau-Dumas B
Biochimie. 1982 Mar;64(3):231-5. doi: 10.1016/s0300-9084(82)80474-4.
Attempts at isolating individual human milk proteins showed that cross interactions made it difficult to obtain of homogeneous components. A new method was devised, based on complete precipitation of milk proteins with saturated ammonium sulphate and progressive solubilization of the precipitate on a column of Sephadex G10 with a linear gradient of ammonium sulphate (from saturation to water). Three fractions were obtained. The first contained lactoferrin, serum albumin, lysozyme and traces of alpha-lactalbumin. Lysozyme could be obtained free from contaminants by chromatography on Ultrogel AcA 54. Lactoferrin and serum albumin coeluting as a single peak, were separated by a further chromatography on DEAE-cellulose. From the other two fractions recovered on Sephadex G10, it should be possible to prepare immunoglobulins, alpha-lactalbumin and the bulk of caseins. The homogeneity of the preparations of lysozyme, lactoferrin and serum albumin was assessed by SDS polyacrylamide gel electrophoresis, acrylamide agarose electrophoresis and immunoelectrophoresis.
对分离个体母乳蛋白质的尝试表明，交叉相互作用使得难以获得均质成分。基于用饱和硫酸铵完全沉淀乳蛋白，并在装有硫酸铵线性梯度（从饱和到水）的Sephadex G10柱上逐步溶解沉淀，设计了一种新方法。得到了三个级分。第一个级分包含乳铁蛋白、血清白蛋白、溶菌酶和微量的α-乳白蛋白。通过在Ultrogel AcA 54上进行色谱分离，可以得到不含污染物的溶菌酶。乳铁蛋白和血清白蛋白作为单一峰共洗脱，通过在DEAE-纤维素上进一步色谱分离。从在Sephadex G10上回收的其他两个级分中，应该有可能制备免疫球蛋白、α-乳白蛋白和大部分酪蛋白。通过SDS聚丙烯酰胺凝胶电泳、丙烯酰胺琼脂糖电泳和免疫电泳评估溶菌酶、乳铁蛋白和血清白蛋白制剂的均质性。
