| Recent interest in conjugated polymers has grown from their demonstrated utility in various "plastic" and/or "molecular" electronic applications to include organic light emitting diodes (OLED's), thin film transistors and photovoltaics. Due to their intrinsically delocalized electronic structure, these same materials show enormous potential as highly responsive optical reporters for chemical and biological interactions. Inter- and intra-chain energy migration, coupled with the formation of strong electrostatic complexes between opposite charged acceptors, allows for extraordinary modulation of their fluorescent response. When these properties are correlated with a specific biological recognition event, the result is a biosensor with optically enhanced or amplified performance. Such features are highly desirable in detection schemes where the target analyte is in limited supply, as is most often the case.; Within these studies we demonstrate how variations in test media composition (i.e. surfactant, buffers, proteins, DNA, etc.) and molecular structure influence those photophysical properties of conjugated polymers related to biosensor design. To this end, both anionic polyphenylenevinylene (PPV) and cationic polyfluorene-cophenylene structures were examined. Model oligomer structures were employed throughout the study for delineating structure-property relationships, as such detailed correlation is inherently more difficult for the less defined polymeric structures (i.e. polydispersity, batch-to-batch variation, purity, etc.). Studies using light scattering and optical spectroscopy highlight the extensive aggregation of these fluorescent, amphiphilic polyelectrolytes in aqueous solution. Variations in chromophore size, charge and concentration provide interesting comparisons in quenching and/or energy transfer processes, as well as, in their interactions with biological molecules.; Ultimately, this information was utilized to develop a novel platform for highly specific and extremely sensitive nucleic acid detection based on the light-harvesting properties of conjugated polymers. Signal transduction within the general assay is controlled by electrostatic interactions and fluorescence resonance energy transfer. The system improves the sensitivity of fluorescent hybridization probes by amplifying their overall emission intensity. Comparisons in sensitivity and simplicity are made to current DNA technologies to emphasize the potential advantages of this new method. Lastly the assay was combined with a common nuclease enzyme to probe for a single base mismatch (or SNP) in human DNA, implicated in a specific neurodegenerative disease. |
