Low cost non-electromechanical technique for the purification of giant unilamellar vesicles

Abstract

Lipid membranes of giant unilamellar vesicles (GUVs) with diameters greater than 10 μm are promising model membranes for investigating the physical and biological properties of the biomembranes of cells. These are extensively used for the study of the interaction of various membrane-active agents, where purified and similar-size oil-free GUVs are necessary. Although the existing membrane filtering method provides the required quality and quantity of GUVs, it includes a relatively expensive double-headed peristaltic pump. In our proposed non-electromechanical technique, gravity is used to maintain the flow of buffer, wherein the flow rate of buffer with the suspension of GUVs is controlled by a locally available low cost roller clamp regulator. We have characterized the results of this non-electromechanical approach in terms of size distribution, average size, flow rate and efficiency for dioleoylphosphatidylglycerol (DOPG)/dioleoylphosphatidylcholine (DOPC)-GUVs prepared by the natural swelling method. The technique purifies the GUVs by removing the non-entrapped solutes at an optimum flow rate 1.0–2.0 mL/min. In addition, it gives similar results to the pump-driven membrane filtering method. Therefore, it might be a cost effective technique for the purification of GUVs without employing any electromechanical devices.