Structure and activity of lipid membrane biosensor surfaces studied with atomic force microscopy and a resonant mirror.
Three variants of the liposome fusion (coalescence) method to produce supported lipid bilayers, containing the ganglioside GM1 on silicon nitride surfaces, were studied. The first procedure involved attachment and fusion of liposomes containing DMPC, GM1 and a small amount of biotinylated lipid (Biotin-LC-DPPE) to a streptavidin coated surface. Direct fusion of liposomes composed of a mixture of DPPC, DPPG, DPPE, GM1 and cholesterol to the surface were the second variant. The final method utilised the second type of liposomes, fused onto a streptavidin layer with a small amount of exposed hydrophobic tails. The methods produced similar lipid layers, but with different ways of attachment to the surface. The binding of cholera toxin B-subunit (CTB) towards these sensor surfaces was measured in a resonant mirror biosensor instrument and the activity and longer-term stability of the layers were examined. The prepared surfaces were also imaged by atomic force microscopy (AFM) in liquid to characterise the topography of the lipid layers. The binding efficiency of CTB towards these surfaces was discussed in terms of lipid fluidity and surface roughness.
Fisher
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DERA Porton Down, Salisbury, Wiltshire, UK.
Tjärnhage
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D015374:Biosensing Techniques / Q000379:methods*; D002772:Cholera Toxin; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes; D008561:Membrane Fusion; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D013499:Surface Properties
D008051:Lipid Bilayers; D008081:Liposomes; D008567:Membranes, Artificial; D002772:Cholera Toxin