Silver nanoparticle-enhanced fluorescence in microtransponder-based immuno- and DNAhybridization assays |
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Authors: | Ji Li Zhuying Wang Ignacy Gryczynski Wlodek Mandecki |
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Institution: | (1) PharmaSeq, Inc, 11 Deer Park Dr., Suite 104, Monmouth Junction, NJ 08852, USA;(2) Present address: GenScript Corporation, 120 Centennial Ave, Piscataway, NJ 08854, USA;(3) Center for Commercialization of Fluorescence Technologies, Department of Molecular Biology and Immunology, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, USA; |
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Abstract: | The aim of this study is to improve assay sensitivity in common solid-phase bioassay configurations as the result of using
silver nanoparticles. The solid phase was provided by numerically indexed, silicon-based electronic chips, microtransponders
(p-Chips) that have previously been used in multiplexed assays. Assay configurations investigated included an ELISA-type immunoassay
and a DNA hybridization assay. The surface of p-Chips was derivatized with the silver island film (SIF) and a polymer, and
then characterized with AFM and SEM. Silver nanoparticle sizes were in the range of 100 to 200 nm. Four fluorophores were
tested for fluorescence enhancement; namely, green fluorescent protein, phycoerythrin, Cy3 and Alexa Fluor 555. We consistently
observed significant fluorescence enhancement and sensitivity improvement in the p-Chip-based assays: the sensitivity in the
cytokine IL-6 immunoassay was 4.3 pg/ml, which represented a 25-fold increase over the method not involving a SIF; and 50 pM
in the hybridization assay, a 38-fold increase. The greatest enhancement was obtained for p-Chip surfaces derivatized first
with the polymer and then coated with SIF. In conclusion, we show that the SIF-p-Chip-based platform is a highly sensitive
method to quantify low-abundance biomolecules in nucleic acid-based assays and immunoassays. |
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