Atger V, Malon D, Bertiere M C, N'Diaye F, Girard-Globa A
INSERM U 286, Faculté Xavier Bichat, Paris, France.
Clin Chem. 1991 Jul;37(7):1149-52.
We used discontinuous gradients of polyacrylamide gel to determine the high-density-lipoprotein (HDL) subfractions HDL2 and HDL3 of serum lipoproteins. Serum (40 microL) prestained with Sudan Black was electrophoresed in cylindrical tubes over successive layers of 3.5%, 6%, 13%, and 17.5% acrylamide gels in a Tris.glycine buffer (3-4 h, 300 V). Very-low- (VLDL) and low-density lipoprotein (LDL) were retained by the 3.5% and 6% gels. HDL2 was concentrated at the interface between the 13% and 17.5% gels, and HDL3 migrated into the 17.5% gel. The distribution between HDL2 and HDL3 was obtained by densitometric scanning. Application of the respective percentages to HDL cholesterol assayed after phosphotung-state-Mg2+ precipitation of VLDL and LDL gave calculated concentrations of HDL2 and HDL3 cholesterol. The calculated values for HDL2 cholesterol were in excellent agreement with those for HDL2 isolated by ultracentrifugation (r = 0.920 for n = 120 sera; differences nonsignificant by Student's paired t-test). Besides being highly discriminating, the method is rapid, easily performed, and economical.
我们使用聚丙烯酰胺凝胶的不连续梯度来测定血清脂蛋白的高密度脂蛋白（HDL）亚组分HDL2和HDL3。用苏丹黑预染的血清（40微升）在Tris-甘氨酸缓冲液中于圆柱形管中在3.5%、6%、13%和17.5%丙烯酰胺凝胶的连续层上进行电泳（3 - 4小时，300伏）。极低密度脂蛋白（VLDL）和低密度脂蛋白（LDL）被3.5%和6%的凝胶保留。HDL2集中在13%和17.5%凝胶的界面处，而HDL3迁移到17.5%的凝胶中。通过光密度扫描获得HDL2和HDL3之间的分布。将各自的百分比应用于在VLDL和LDL经磷钨酸盐 - Mg2 +沉淀后测定的HDL胆固醇，得出HDL2和HDL3胆固醇的计算浓度。HDL2胆固醇的计算值与通过超速离心分离的HDL2的值高度一致（n = 120份血清时r = 0.920；经学生配对t检验差异不显著）。该方法除了具有高度区分性外，还快速、易于操作且经济。
