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

✍ Scribed by Michel Ollivon; Anne Walter; Robert Blumenthal

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 985 KB
• Volume: 152
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90408-2

✦ Synopsis

The ability of an HPLC gel exclusion column (TSK G6OOOPW) to separate lipid vesicles, viruses. and biological vesicles according to size was tested and compared with separations on Sephacryl SIOOO. 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 G60OOPW column wasable to separate larger particles (>500 nm) than the Sephacryl SIOOO. and, when used in combination with the TSK G5OOOPW column, gave more discrete separations of smaller particles (IO to 30 nm diameter). Moreover, the HPLC columns can be run significantly faster (IO-20 min vs several hours) and give more precise results than Sephacryl SIOOO. Therefore, HPLC using a G6OOOPW column alone, or in combination with a G5OOOPW column, provides a rapid and accurate means of sizing and selecting specifically sized biological and artificial vesicles. 8 1986 Academic press. hc.