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Silver nanoparticle-enhanced fluorescence in microtransponder-based immuno- and DNAhybridization assays

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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.

Green fluorescent protein (GFP) and Alexa Fluor 555 deposited on microtransponders in the presence or absence of the silver island film.

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Acknowledgments

We thank Richard G. Morris and Maryann Gruda (PharmaSeq, Inc.) for insightful discussions regarding the experiment setup and reading of the manuscript, Irina Akopova (University of North Texas Health Science Center) for collection of AFM images and Tanya Shtoyko (University of Texas at Tyler) for performing SEM measurements.

This work was supported by a grant from National Institutes of Health [CA132547 to WM].

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Author notes

  1. Zhuying Wang

    Present address: GenScript Corporation, 120 Centennial Ave, Piscataway, NJ, 08854, USA

Authors and Affiliations

  1. PharmaSeq, Inc, 11 Deer Park Dr., Suite 104, Monmouth Junction, NJ, 08852, USA

    Ji Li, Zhuying Wang & Wlodek Mandecki

  2. 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

    Ignacy Gryczynski

Authors
  1. Ji Li
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  2. Zhuying Wang
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  3. Ignacy Gryczynski
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  4. Wlodek Mandecki
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Correspondence to Wlodek Mandecki.

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Li, J., Wang, Z., Gryczynski, I. et al. Silver nanoparticle-enhanced fluorescence in microtransponder-based immuno- and DNAhybridization assays. Anal Bioanal Chem 398, 1993–2001 (2010). https://doi.org/10.1007/s00216-010-4108-7

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  • DOI: https://doi.org/10.1007/s00216-010-4108-7

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