Burstein M, Scholnick H R, Morfin R
J Lipid Res. 1970 Nov;11(6):583-95.
Procedures are described for the isolation of lipoproteins from human serum by precipitation with polyanions and divalent cations. A mixture of low and very low density lipoproteins can be prepared without ultracentrifugation by precipitation with heparin and either MnCl(2) alone or MgCl(2) plus sucrose. In both cases the precipitation is reversible, selective, and complete. The highly concentrated isolated lipoproteins are free of other plasma proteins as judged by immunological and electrophoretic methods. The low density and very low density lipoproteins can then be separated from each other by ultracentrifugation. The advantage of the method is that large amounts of lipoproteins can be prepared with only a single preparative ultracentrifugation. Polyanions other than heparin may also be used; when the precipitation of the low and very low density lipoproteins is achieved with dextran sulfate and MnCl(2), or sodium phosphotungstate and MgCl(2), the high density lipoproteins can subsequently be precipitated by increasing the concentrations of the reagents. These lipoproteins, containing small amounts of protein contaminants, are further purified by ultracentrifugation at d 1.22. With a single preparative ultracentrifugation, immunologically pure high density lipoproteins can be isolated from large volumes of serum.
本文描述了通过用聚阴离子和二价阳离子沉淀从人血清中分离脂蛋白的方法。通过用肝素和单独的MnCl₂或MgCl₂加蔗糖沉淀，可以在不进行超速离心的情况下制备低密度和极低密度脂蛋白的混合物。在这两种情况下，沉淀都是可逆的、选择性的且完全的。通过免疫和电泳方法判断，高度浓缩的分离脂蛋白不含其他血浆蛋白。然后可以通过超速离心将低密度和极低密度脂蛋白彼此分离。该方法的优点是仅通过一次制备性超速离心就可以制备大量脂蛋白。也可以使用肝素以外的聚阴离子；当用硫酸葡聚糖和MnCl₂或磷钨酸钠和MgCl₂实现低密度和极低密度脂蛋白的沉淀时，随后可以通过增加试剂浓度来沉淀高密度脂蛋白。这些含有少量蛋白质污染物的脂蛋白通过在d 1.22下超速离心进一步纯化。通过一次制备性超速离心，可以从大量血清中分离出免疫纯的高密度脂蛋白。
