Groot P H, Scheek L M, Havekes L, van Noort W L, van't Hooft F M
J Lipid Res. 1982 Dec;23(9):1342-53.
A method was developed for the separation of the high density lipoprotein subclasses HDL2 and HDL3 from human serum. Six serum samples are fractionated in a single-step ultracentrifugal procedure using the Beckman (SW-40) swinging bucket rotor. The method is based on a difference in flotation rate of the high density lipoprotein subclasses. Separation of HDL2 and HDL3 is accomplished by a discontinuous NaBr density gradient applied on top of 2 ml of serum brought to a density of 1.40 g/ml. After centrifugation, high density lipoprotein subclass profiles were obtained using a specially designed gradient fractionator. Contamination of the isolated high density lipoprotein subclasses by serum albumin or by apolipoprotein B-containing lipoproteins was minimal while only a slight overlap between the HDL2 and HDL3 profiles was observed. Chemical and immunochemical analyses of the high density lipoprotein subclasses isolated by the present method were in close agreement with the results obtained by rate-zonal density gradient ultracentrifugation in zonal rotors (Patsch, et al. 1980. J. Biol. Chem. 255: 3178-3185). The major advantage of the method presented in this paper as compared with the zonal rotor method is the possibility to analyze as many as six serum samples simultaneously.
已开发出一种从人血清中分离高密度脂蛋白亚类HDL2和HDL3的方法。使用贝克曼（SW - 40）摆动式转头，通过单步超速离心程序对六个血清样本进行分级分离。该方法基于高密度脂蛋白亚类在漂浮速率上的差异。HDL2和HDL3的分离是通过在2 ml密度调至1.40 g/ml的血清顶部施加不连续的溴化钠密度梯度来实现的。离心后，使用专门设计的梯度分级收集器获得高密度脂蛋白亚类图谱。分离出的高密度脂蛋白亚类受血清白蛋白或含载脂蛋白B的脂蛋白污染极少，而HDL2和HDL3图谱之间仅观察到轻微重叠。用本方法分离出的高密度脂蛋白亚类的化学和免疫化学分析结果与在区带转头中进行速率区带密度梯度超速离心所获得的结果密切相符（帕奇等人，1980年。《生物化学杂志》255：3178 - 3185）。与区带转头法相比，本文所述方法的主要优点是能够同时分析多达六个血清样本。
