Supramolecular interfacial architectures for optical biosensing with surface plasmons

We describe several approaches to design, synthesize and assemble supramolecular (bio-)functional interfacial architectures for applications in optical biosensing using, in particular, surface plasmon field-enhanced fluorescence spectroscopy (SPFS). Firstly, we discuss the build-up of an interfacial catcher probe layer for surface-hybridization studies with PCR amplicons. The well-established biotin-streptavidin coupling scheme is used to assemble a peptide nucleic acid (PNA) probe matrix. SPFS allows then for a very detailed and quantitative evaluation of the kinetics and affinities of the association and dissociation reactions between these catcher oligonucleotide strands and chromophore-labeled PCR (125 bp) strands from solution. The second example concerns the study of protein binding using an ELISA-analogue sandwich approach: a primary antibody against the prostate-specific antigen (PSA) used in these examples is coupled to a dextran binding matrix at the sensor surface via EDC/[NHS-coupling. The detection limits for PSA are then evaluated using a 2-step- or 1-step-antigen/secondary antibody strategy by monitoring the fluorescence intensity emitted from chromophore-labels covalently bound to the secondary antibody. The final system that we describe involves a novel model membrane system, i.e., a tethered bimolecular lipid membrane (tBLM). Reconstitution of integrin receptors then allows for a quantitative study of the binding of fluorophore-labeled collagen fragments to the membrane-based integrin receptors. (C) 2004 Elsevier B.V. All rights reserved.