Tetaz T, Kecorius E, Grego B, Fidge N
Baker Medical Research Institute, Melbourne, Victoria, Australia.
J Chromatogr. 1990 Jul 6;511:147-53. doi: 10.1016/s0021-9673(01)93280-7.
A method has been developed for the rapid separation of the medium-molecular-weight apolipoproteins A-IV, A-I and E by high-performance liquid chromatography. Separations were achieved using a commercially available column of very low hydrophobicity (TSK Phenyl-5PW) in the reversed-phase mode rather than the conventional mode of hydrophobic interaction. Delipidated apolipoproteins were dissolved in 20 mM orthophosphoric acid (pH 2.3), applied to the column which was pre-equilibrated with the same buffer, and eluted with an increasing gradient of acetonitrile. Purified apolipoproteins were identified by a combination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis, amino acid analysis and N-terminal sequence analysis. In one step the method can be used to separate the major human chylomicron apolipoproteins A-IV, A-I and E, following preliminary removal of apolipoprotein A-II and the C apolipoproteins by size-exclusion chromatography.
已开发出一种通过高效液相色谱快速分离中等分子量载脂蛋白A-IV、A-I和E的方法。分离是在反相模式下使用市售的极低疏水性柱（TSK苯基-5PW）实现的，而不是传统的疏水相互作用模式。脱脂载脂蛋白溶解在20 mM正磷酸（pH 2.3）中，应用于用相同缓冲液预平衡的柱上，并用乙腈梯度增加进行洗脱。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、氨基酸分析和N端序列分析相结合的方法鉴定纯化的载脂蛋白。在初步通过尺寸排阻色谱法去除载脂蛋白A-II和C载脂蛋白后，该方法可一步用于分离主要的人乳糜微粒载脂蛋白A-IV、A-I和E。
