Carroll R M, Rudel L L
J Lipid Res. 1983 Feb;24(2):200-7.
Plasma lipoproteins were isolated at d less than 1.225 g/ml from nonhuman primates of three species, cynomolgus, rhesus, and African green (vervet) monkeys. Individual lipoprotein classes were separated by high performance gelfiltration chromatography and low density lipoprotein (LDL) molecular weight was determined. A comparison was made using column configurations including TSK 3000 SW, 4000 SW, and 5000 PW columns. Due to its relative simplicity, stability, and economy, a single 5000 PW column was selected for most of the work. The recovery of lipoprotein cholesterol from the column averaged 91 +/- 2.5%. A comparison of the immunologic, chemical, and electrophoretic properties of high density lipoproteins (HDL) and LDL isolated by this technique with those of HDL and LDL isolated by conventional agarose column chromatography indicated that lipoproteins isolated by high performance gel-filtration chromatography were intact and reasonably free of cross contamination. A standard preparation of 125I-labeled LDL was added to the d less than 1.225 g/ml lipoprotein fraction just prior to separation and a relative size index, r1, was determined. When r1 values for a large number of samples were compared with the log of the LDL molecular weight (determined by agarose column chromatography) a linear relationship was found with a correlation coefficient, r = 0.85. The regression equation for this relationship could be used to calculate LDL molecular weights from the r1 value. These values agreed with LDL molecular weight determined by flotation equilibrium analysis in the analytical ultracentrifuge. We conclude that high performance gel-filtration chromatography using the TSK 5000 PW column provides an analytical and preparative technique for simultaneous separation of individual lipoproteins and determination of LDL molecular weight.
从食蟹猴、恒河猴和非洲绿猴（草原猴）这三种非人灵长类动物中分离出密度小于1.225 g/ml的血浆脂蛋白。通过高效凝胶过滤色谱法分离各个脂蛋白类别，并测定低密度脂蛋白（LDL）的分子量。使用包括TSK 3000 SW、4000 SW和5000 PW柱在内的柱配置进行了比较。由于其相对简单、稳定且经济，大部分工作选用了单一的5000 PW柱。从柱中回收的脂蛋白胆固醇平均为91±2.5%。将通过该技术分离的高密度脂蛋白（HDL）和LDL的免疫、化学和电泳特性与通过传统琼脂糖柱色谱法分离的HDL和LDL的特性进行比较，结果表明通过高效凝胶过滤色谱法分离的脂蛋白完整且基本无交叉污染。在分离前，将125I标记的LDL标准制剂添加到密度小于1.225 g/ml的脂蛋白组分中，并测定相对大小指数r1。当将大量样品的r1值与LDL分子量的对数（通过琼脂糖柱色谱法测定）进行比较时，发现存在线性关系，相关系数r = 0.85。该关系的回归方程可用于根据r1值计算LDL分子量。这些值与在分析超速离心机中通过浮选平衡分析测定的LDL分子量一致。我们得出结论，使用TSK 5000 PW柱的高效凝胶过滤色谱法提供了一种用于同时分离单个脂蛋白和测定LDL分子量的分析和制备技术。
