通过高效液相色谱法对脂质体、生物囊泡和病毒进行尺寸测定与分离。

Sizing and separation of liposomes, biological vesicles, and viruses by high-performance liquid chromatography.

作者信息

Ollivon M, Walter A, Blumenthal R

出版信息

Anal Biochem. 1986 Feb 1;152(2):262-74. doi: 10.1016/0003-2697(86)90408-2.

Abstract

The ability of an HPLC gel exclusion column (TSK G6000PW) to separate lipid vesicles, viruses, and biological vesicles according to size was tested and compared with separations on Sephacryl S1000. The columns were calibrated using vesicular Stokes radii determined by quasielastic light scattering. The vesicles separated according to size on both types of column and remained intact during elution. Viruses of known diameters and clathrin-coated vesicles were also eluted as a function of size. The TSK G6000PW column was able to separate larger particles (greater than 500 nm) than the Sephacryl S1000, and, when used in combination with the TSK G5000PW column, gave more discrete separations of smaller particles (10 to 30 nm diameter). Moreover, the HPLC columns can be run significantly faster (10-20 min vs several hours) and give more precise results than Sephacryl S1000. Therefore, HPLC using a G6000PW column alone, or in combination with a G5000PW column, provides a rapid and accurate means of sizing and selecting specifically sized biological and artificial vesicles.

摘要

测试了高效液相色谱凝胶排阻柱（TSK G6000PW）根据大小分离脂质体、病毒和生物小泡的能力，并与在Sephacryl S1000上的分离效果进行了比较。使用通过准弹性光散射测定的囊泡斯托克斯半径对柱子进行校准。两种类型的柱子上的囊泡均根据大小进行分离，并且在洗脱过程中保持完整。已知直径的病毒和网格蛋白包被的小泡也根据大小被洗脱。TSK G6000PW柱能够分离比Sephacryl S1000更大的颗粒（大于500 nm），并且当与TSK G5000PW柱联合使用时，能对更小的颗粒（直径10至30 nm）进行更离散的分离。此外，与Sephacryl S1000相比，高效液相色谱柱的运行速度明显更快（10 - 20分钟对几个小时），且结果更精确。因此，单独使用G6000PW柱或与G5000PW柱联合使用的高效液相色谱法提供了一种快速且准确的方法，用于对特定大小的生物和人工小泡进行大小测定和选择。