title: A membrane filtering method for the purification of giant unilamellar vesicles.
abstract: The use of giant unilamellar vesicles (GUVs) for investigating the properties of biomembranes is advantageous compared to the use of small-sized vesicles such as large unilamellar vesicles (LUVs). Experimental methods using GUVs, such as the single GUV method, would benefit if there was a methodology for obtaining a large population of similar-sized GUVs composed of oil-free membranes. We here describe a new membrane filtering method for purifying GUVs prepared by the natural swelling method and demonstrate that, following purification of GUVs composed of dioleoylphosphatidylglycerol (DOPG)/dioleoylphosphatidylcholine (DOPC) membranes suspended in a buffer, similar-sized GUVs with diameters of 10-30 μm are obtained. Moreover, this method enabled GUVs to be separated from water-soluble fluorescent probes and LUVs. These results suggest that the membrane filtering method can be applied to GUVs prepared by other methods to purify larger-sized GUVs from smaller GUVs, LUVs, and various water-soluble substances such as proteins and fluorescent probes. This method can also be used for concentration of dilute GUV suspensions.
authors.affiliation: Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, Suruga-ku, Shizuoka, Japan.
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Tamba
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Terashima
authors.forename: Masahito
authors.initials: M
authors.lastname: Yamazaki
mesh_terms: D005374:Filtration / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D053835:Unilamellar Liposomes / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D005456:Fluorescent Dyes; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D053835:Unilamellar Liposomes; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol; C017251:1,2-oleoylphosphatidylcholine