Meyer B, Kecorius E, Barter P, Fidge N, Tetaz T
Baker Medical Research Institute, Prahran, Victoria, Australia.
J Chromatogr. 1991 Mar 1;540(1-2):386-91. doi: 10.1016/s0021-9673(01)88827-0.
Apolipoproteins A-IV, A-I and E from rat high-density lipoprotein (HDL) were successfully purified by reversed-phase high-performance liquid chromatography (RP-HPLC), using a method which we have previously developed for the separation of apolipoproteins A-IV, A-I and E from human lymph chylomicrons [T. Tetaz, E. Kecorius, B. Grego and N. Fidge, J. Chromatogr., 511 (1990) 147]. Since analytical-scale RP-HPLC indicated that the C apolipoproteins from rat HDL coeluted with both apo A-IV and apo A-I, delipidated rat HDL was first subjected to preparative-scale size-exclusion HPLC (HPSEC) on a Serva Si300 column, which effectively separated the C apolipoproteins from all but apolipoprotein E. Fractions from HPSEC which were enriched for apolipoproteins A-IV, A-I or E were directly applied to RP-HPLC on a TSK Phenyl-5PW column. This procedure yielded fractions containing apolipoproteins A-IV, A-I or E which were pure as assessed by N-terminal sequencing and silver staining of sodium dodecyl sulphate-polyacrylamide gels.
利用我们之前开发的从人淋巴乳糜微粒中分离载脂蛋白A-IV、A-I和E的方法，通过反相高效液相色谱（RP-HPLC）成功纯化了大鼠高密度脂蛋白（HDL）中的载脂蛋白A-IV、A-I和E [T. Tetaz, E. Kecorius, B. Grego和N. Fidge, J. Chromatogr., 511 (1990) 147]。由于分析规模的RP-HPLC表明大鼠HDL中的C载脂蛋白与载脂蛋白A-IV和载脂蛋白A-I共洗脱，因此首先将脱脂的大鼠HDL在Serva Si300柱上进行制备规模的尺寸排阻高效液相色谱（HPSEC），该柱有效地将C载脂蛋白与除载脂蛋白E之外的所有载脂蛋白分离。HPSEC中富含载脂蛋白A-IV、A-I或E的馏分直接应用于TSK Phenyl-5PW柱上的RP-HPLC。通过N端测序和十二烷基硫酸钠-聚丙烯酰胺凝胶的银染评估，该方法得到了含有纯的载脂蛋白A-IV、A-I或E的馏分。
