Hydroxylamine amplified gold nanoparticle-based aptameric system for the highly selective and sensitive detection of platelet-derived growth factor

This study developed a sensitive and selective aptamer-based chemiluminescent (CL) method for the determination of platelet-derived growth factor (PDGF)-BB using hydroxylamine enlarged gold nanoparticles (Au NPs). Rabbit anti-human PDGF-BB polyclonal antibody was covalently coupled on the 96-well plate that offers reactive N-oxysuccinimide ester (referred to as NOS group) surface. In the presence of target protein, the biotinylated aptamer was captured on the 96-well plate forming an antibody/PDGF-BB/biotinylated aptamer sandwiched complex, which was followed by the assembly of streptavidin coated Au NPs (streptavidin-gold). Au NPs assembled on the surface of 96-well plate reacted with HAuCl4 and NH2OH, which enabled the catalytic deposition of gold metal onto the Au NPs surfaces. A huge number of Au3+ ions were released from the hydroxylamine enlarged Au NPs after oxidative gold metal dissolution, which was determined by a simple and sensitive luminol CL reaction. The results showed that the detection limit of the assay is 60 pM of PDGF-BB (corresponding to 6 fmol in a 100 mu L volume), which compares favorably with those of other PDGF-BB detection techniques. In addition, this aptameric CL biosensor demonstrated extraordinary specificity. And PDGF-BB has been determined in diluted serum indicating the applicability of this assay. (C) 2012 Elsevier B.V. All rights reserved.