Sensitive detection of protein adsorption to supported lipid bilayers by frequency-dependent capacitance measurements and microelectrophoresis.
In the first part, we report experiments which enable the sensitive detection of protein adsorption to lipid bilayers deposited onto chromium electrodes on glass substrates by frequency-dependent capacitance measurements. The sensitivity of the present type of sensor (better than 0.3 nm average protein layer thickness) is at least equivalent to that of ellipsometry. A high specific resistance of the supported bilayer of (1-5).10(5) omega.cm2 is achieved by deposition of a tightly packed (crystalline) cadmium arachidate monolayer in contact with the substrate, whereas the outer monolayer can be more loosely packed (fluid phase or state of fluid-solid coexistence) which is essential for the incorporation of receptors. In the present work, charged lipids are incorporated as nonspecific receptors for polylysine and cytochrome c. The capacitance measurements provide a very sensitive test of the tightness and the long-time stability of the supported bilayers and, in combination with ellipsometric thickness measurements, enable estimations of dielectric properties of protein layers (such as the permittivity). In the second part, we report first electrophoresis experiments in asymmetric bilayers on substrates which enable simultaneous measurements of lateral diffusion coefficients and frictional coefficients between monolayers. The potential application of the electrophoretic effect for the differentiation between different receptors and the amplification of signals in biosensors is discussed.
Stelzle
M
M
Physik-Department, Technische Universität München, F.R.G.
Sackmann
E
E
D015060:1,2-Dipalmitoylphosphatidylcholine; D000327:Adsorption; D003574:Cytochrome c Group / Q000032:analysis; D004134:Dimyristoylphosphatidylcholine; D004553:Electric Conductivity; D004563:Electrochemistry; D004586:Electrophoresis / Q000379:methods; D008051:Lipid Bilayers; D008832:Microchemistry; D011107:Polylysine / Q000032:analysis; D011506:Proteins / Q000032:analysis
D003574:Cytochrome c Group; D008051:Lipid Bilayers; D011506:Proteins; D011107:Polylysine; D015060:1,2-Dipalmitoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine