The University of Southampton

ProteoGUV platforms

GUV electroformation
Date:
2014-2018
Theme:
Bionanotechnology and Biosensors

Over the last decade, giant unilamellar vesicles (GUVs) have emerged as a valuable tool for the study of membrane domains, receptor and ion channel function, membrane morphology, and also as models for 'proto-cells'. GUVs are liposomes (lipid bilayers enclosing an aqueous volume) with a diameter of tens of micrometers. The large size of these structures enables optical imaging of membrane domains and of vesicle morphology as well as clamping of the vesicle with micropipettes, while the diffusion of lipids and membrane proteins is not restricted because there is no solid support layer. GUVs have become a popular research topic with the introduction of the electroformation protocol by Angelova and Dimitrov, who showed that some lipids, as a dried film on electrodes, formed GUVs when rehydrated with a salt-less or low-salt solution while an AC field is applied. This method, which is postulated to involve localized electroswelling of lipid bilayers, has been gradually optimized to allow GUV formation with a wider variety of lipid species and at more physiological (~100 mM) salt concentrations. The key change is that the lipids are deposited on the electrodes as small unilamellar liposomes (SUVs), which are partially but not completely dried prior to rehydration in the AC field. More recently, it has been demonstrated that the use of protein-containing liposomes (proteoSUVs) enables the formation of protein-containing GUVs. In this project, we will systematically explore this exciting development to derive standard procedures for microsystem-enabled semi-automated proteoGUV formation, and employ ion channel GUVs for drug screening with patch clamp methods.

Primary investigators

Secondary investigator

  • Hang Lu

Partner

  • Dr Ita O'Kelly (Medicine)

Associated research group

  • Electronics and Electrical Engineering
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