Choice E, Ayyobi A F, Pritchard P H, Madden T D
Department of Pharmacology & Therapeutics, University of British Columbia, 2176 Health Sciences Mall, Vancouver, British Columbia, V6T 1Z3, Canada.
Anal Biochem. 1999 May 15;270(1):1-8. doi: 10.1006/abio.1999.4049.
We describe an efficient method for separating liposomes (large unilamellar vesicles, 120-150 nm diameter) from plasma lipoproteins employing fast protein liquid chromatography (FPLC). This method resolves very low density lipoprotein (VLDL), low-density lipoprotein, high-density lipoprotein, and other plasma components. Selective detection of liposomes (large unilamellar vesicles, 120-150 nm diameter) was achieved using either radiolabeled or fluorescent lipid probes. The liposomes were found to coelute with the earliest FPLC-eluting lipoprotein fraction, VLDL. The remaining plasma lipoprotein and protein components eluted at later times and were resolved from liposomes and VLDL. In order to separate VLDL from liposomes, we selectively precipitated the VLDL fraction from plasma using tungstophosphoric acid and magnesium chloride, prior to separation by FPLC. Furthermore, we demonstrate that this technique can be used to separate liposomes from lipoproteins in plasma samples collected after intravenous administration of liposomes to mice. This technique has wide application in studies of liposome stability in blood and, in particular, for the characterization of liposomal drug carrier systems.
我们描述了一种利用快速蛋白质液相色谱法（FPLC）从血浆脂蛋白中分离脂质体（直径120 - 150 nm的大单层囊泡）的有效方法。该方法可分离极低密度脂蛋白（VLDL）、低密度脂蛋白、高密度脂蛋白及其他血浆成分。使用放射性标记或荧光脂质探针可实现对脂质体（直径120 - 150 nm的大单层囊泡）的选择性检测。发现脂质体与最早从FPLC洗脱的脂蛋白组分VLDL一同洗脱。其余血浆脂蛋白和蛋白质组分在较晚时间洗脱，并与脂质体和VLDL分离。为了从脂质体中分离VLDL，在通过FPLC分离之前，我们使用磷钨酸和氯化镁从血浆中选择性沉淀VLDL组分。此外，我们证明该技术可用于从小鼠静脉注射脂质体后采集的血浆样品中分离脂质体和脂蛋白。该技术在脂质体在血液中的稳定性研究中具有广泛应用，尤其适用于脂质体药物载体系统的表征。
