Novel plasmonic field-enhanced nanoassay for trace detection of proteins

Recently discovered effects of plasmonic field on molecular fluorescence offer new insights into the optical interactions at the nanoscale which can help solving problems encountered in widely applied fluorescent labeling of biomolecules for studying life processes in biomedicine and pharmacy. In this work, we have focused on exploring a novel sensitivity-enhancing phenomenon based on protein modulation of plasmoncontrolled fluorescence. We have demonstrated that a protein (cytochrome c (Cytc c) or bovine serum albumin (BSA)) can be employed to gate fluorescence resonance energy transfer occurring from a fluorescein isothiocyanate fluorescent dye to plasmonic citrate-capped gold nanoparticles. By applying plasmonic field gated by protein, facilitated by the formation of multi-shell nanoparticles, (AuNP@Cit/Cytc-FITC or AuNP@Cit/BSA-FITC), low limits of detection for Cyt c (LOD=370 pM) and for BSA (LOD= 1.8 nM) have been achieved even for large fluorophore:protein ratios, up to 30:1 (over-labeling), normally plagued with energy migration and background fluorescence problems. Control experiments confirming adsorption of proteins on AuNPs have been performed using light scattering and piezometric techniques. The proposed nanoassay may be applied in microanalysis of trace amounts of proteins, e.g. in microfluidic devices. (C) 2013 Elsevier B.V. All rights reserved.