A dry membrane protection technique to allow surface acoustic wave biosensor measurements of biological model membrane approaches.
Model membrane approaches have attracted much attention in biomedical sciences to investigate and simulate biological processes. The application of model membrane systems for biosensor measurements is partly restricted by the fact that the integrity of membranes critically depends on the maintenance of an aqueous surrounding, while various biosensors require a preconditioning of dry sensors. This is for example true for the well-established surface acoustic wave (SAW) biosensor SAM®5 blue. Here, a simple drying procedure of sensor-supported model membranes is introduced using the protective disaccharide trehalose. Highly reproducible model membranes were prepared by the Langmuir-Blodgett technique, transferred to SAW sensors and supplemented with a trehalose solution. Membrane rehydration after dry incorporation into the SAW device becomes immediately evident by phase changes. Reconstituted model membranes maintain their full functionality, as indicated by biotin/avidin binding experiments. Atomic force microscopy confirmed the morphological invariability of dried and rehydrated membranes. Approximating to more physiological recognition phenomena, the site-directed immobilization of the integrin VLA-4 into the reconstituted model membrane and subsequent VCAM-1 ligand binding with nanomolar affinity were illustrated. This simple drying procedure is a novel way to combine the model membrane generation by Langmuir-Blodgett technique with SAW biosensor measurements, which extends the applicability of SAM®5 blue in biomedical sciences.
Pharmaceutical Chemistry, Rheinische Friedrich-Wilhelms-Universität Bonn, An der Immenburg 4, Bonn D-53121, Germany. kchrist@uni-bonn.de.
Katrin
K
Reder-Christ
Patrick
P
Schmitz
Marian
M
Bota
Ursula
U
Gerber
Hildegard
H
Falkenstein-Paul
Christian
C
Fuss
Marius
M
Enachescu
Gerd
G
Bendas
D032701:Biomimetics / Q000295:instrumentation*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation*; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D055617:Micro-Electrical-Mechanical Systems / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008051:Lipid Bilayers; D008567:Membranes, Artificial