S-layer based concept for the development of nano-structured biosensors

The challenge for new sensor systems is to make them smaller and to achieve higher specifity and sensitivity to one analyte. Most promising, therefore, are bio molecules due to their typical sizes in the nano-meter range, self-assembly properties and the high affinity of biological binding molecules. Here we present a method to construct a nano-structured sensor device, using surface layer (S-layer) proteins as templates, aptamers (short oligonucleotides) as receptors and a dye pair for fluorescence resonance energy transfer (FRET) to detect the binding of one analyte . S-layer proteins are structural proteins, forming the outermost cell envelope of numerous bacteria and almost all archaea. They feature a lot of functions for the microbial cell such as protection, adhesion, filtration or framework. Their oblique, square or hexagonal structure, high content of regular arranged functional groups and the ability of the monomers to self-assembly in aqueous solution qualifies them for various nanotechnological purposes. The functional groups of the side chains of the S-layer proteins can be used for the sequential coupling of aptamers and organic or inorganic fluorescent dyes. In combination with an optical device for detection, a nano-structured sensor system is constructible. Aptamers were chosen as receptor system due to their high specificity, comparable to that of antibodies, and their higher stability against environmental changes. Currently we are working on a sensor system which is able to detect pharmaceuticals in water. Therefore different S-layer, aptamer and dye conformations will be tested to optimize the binding of a target molecule and to maximize the interference of the initial FRET between the two fluorescent dyes. It is expected to generate an optical signal that will allow the detection of very low analyte concentrations. An additional aim for the future is to bind two or more different aptamers to the S-layers, to develop a multiple detection system. Beside the construction of sensory devices, the aptamer-S-layers combination can be used for the construction of filter materials for the specific binding and degradation of toxic organic substances in water. For this purpose, photocatalytic active nanoparticles and aptamers are regularly arranged on the S-layer protein. Bound organic substances can be eliminated by the formation of reactive oxygen species induced by light irradiation.