A novel method for the rapid separation of plasma lipoproteins using self-generating gradients of iodixanol
A novel method for the rapid separation of plasma lipoproteins using self-generating gradients of iodixanol
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DOI：
10.1016/0021-9150(96)05797-8
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发表时间：
1996-07-01
期刊：
影响因子：
5.3
通讯作者：
Billington, D
中科院分区：
文献类型：
--
作者：
Graham, JM;Higgins, JA;Billington, D
我们描述了一种快速分离血浆脂蛋白的新方法，该方法利用了一种新的非离子、碘化、密度梯度介质碘克沙醇（商品名为OptiprepTM）。该方法简单：将血浆或血清与碘克沙醇混合，然后在垂直或接近垂直的转子中离心。VLDL、LDL和HDL的分离可以在3小时内实现，并且脂蛋白级分在密度和组成上与使用常规盐基梯度制备的那些相当。每类脂蛋白可以在单个组分中去除，或者可以使用梯度分馏器获得脂蛋白分布的概况。由于培养基是惰性的，因此可以通过琼脂糖凝胶电泳分析来自梯度的级分，或通过SDS-PAGE测定脂质含量或载脂蛋白组成，而无需除去碘克沙醇。几个梯度组分中HDL和LDL的电泳迁移率的微小差异表明这些类别可能会发生细分。该方法简便、快速、通用性强，可用于脂蛋白的制备和临床实验室脂蛋白谱的分析。
We describe a new method for the rapid fractionation of plasma lipoproteins, which makes use of a new non-ionic, iodinated, density gradient medium, iodixanol, commercially available as Optiprep(TM). The method is simple: plasma or serum is mixed with iodixanol followed by centrifugation in a vertical or near vertical rotor. Separation of VLDL, LDL and HDL can be achieved in 3 h and the lipoprotein fractions are comparable in density and composition with those prepared using conventional salt based gradients. Each class of lipoprotein can be removed in a single fraction, or a profile of lipoprotein distribution can be obtained using a gradient fractionator. Because the medium is inert, fractions from the gradient can be analysed by agarose gel electrophoresis or assayed for lipid content or apolipoprotein composition by SDS-PAGE without removing the iodixanol. Small differences in electrophoretic mobility of HDL and LDL across several gradient fractions suggest that subfractionation of these classes may occur. The new method is simple, rapid and versatile with potential application for preparation of lipoproteins and for analysis of lipoprotein profiles in the research or clinical laboratory.