Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces

Evgenia Matveeva; Zygmunt Gryczynski; Joanna Malicka; Ignacy Gryczynski; Joseph R. Lakowicz

doi:10.1016/j.ab.2004.08.034

Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces

Evgenia Matveeva, Zygmunt Gryczynski, Joanna Malicka, Ignacy Gryczynski, Joseph R. Lakowicz

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111 Scopus citations

Abstract

We present a generic immunoassay platform that uses enhanced total internal reflection fluorescence in the proximity of silver island films (SIFs), a surface coating consisting of metal (silver) particles. This platform is used with a model immunoassay where a protein antigen, rabbit immunoglobulin G, was immobilized on the SIF-coated glass surface. The signal from a fluorescent dye-labeled anti-rabbit antibody binding to the surface antigen was detected; different color dyes have been tested. Close placement of the fluorophore to surface-bound silver nanostructures results in dramatic signal enhancement (up to 40-fold) on the SIFs as compared with the glass slides. Use of the total internal reflection mode of excitation has significant advantages (over classic front-face excitation) for practical assay development. The limited evanescent wave excitation volume makes it possible to minimize the background signal and use the immunoassay with no need for any washing steps.

Original language	English
Pages (from-to)	303-311
Number of pages	9
Journal	Analytical Biochemistry
Volume	334
Issue number	2
DOIs	https://doi.org/10.1016/j.ab.2004.08.034
State	Published - 15 Nov 2004

Keywords

Enhanced fluorescence
Fluorescence immunoassay
Immunoglobulin
Nano-size metallic particles
Silver island films

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10.1016/j.ab.2004.08.034

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title = "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces",

abstract = "We present a generic immunoassay platform that uses enhanced total internal reflection fluorescence in the proximity of silver island films (SIFs), a surface coating consisting of metal (silver) particles. This platform is used with a model immunoassay where a protein antigen, rabbit immunoglobulin G, was immobilized on the SIF-coated glass surface. The signal from a fluorescent dye-labeled anti-rabbit antibody binding to the surface antigen was detected; different color dyes have been tested. Close placement of the fluorophore to surface-bound silver nanostructures results in dramatic signal enhancement (up to 40-fold) on the SIFs as compared with the glass slides. Use of the total internal reflection mode of excitation has significant advantages (over classic front-face excitation) for practical assay development. The limited evanescent wave excitation volume makes it possible to minimize the background signal and use the immunoassay with no need for any washing steps.",

keywords = "Enhanced fluorescence, Fluorescence immunoassay, Immunoglobulin, Nano-size metallic particles, Silver island films",

author = "Evgenia Matveeva and Zygmunt Gryczynski and Joanna Malicka and Ignacy Gryczynski and Lakowicz, {Joseph R.}",

note = "Funding Information: This work was supported by the National Center for Research Resource (RR-08119), the National Institute of Biomedical Imaging and Bioengineering (EB-1690), and Philip Morris USA Inc. and Philip Morris International. The authors are grateful to Mikhail Matveev for writing the software for enhancement calculations. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2004",

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doi = "10.1016/j.ab.2004.08.034",

language = "English",

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AU - Matveeva, Evgenia

AU - Gryczynski, Zygmunt

AU - Malicka, Joanna

AU - Gryczynski, Ignacy

AU - Lakowicz, Joseph R.

N1 - Funding Information: This work was supported by the National Center for Research Resource (RR-08119), the National Institute of Biomedical Imaging and Bioengineering (EB-1690), and Philip Morris USA Inc. and Philip Morris International. The authors are grateful to Mikhail Matveev for writing the software for enhancement calculations. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2004/11/15

Y1 - 2004/11/15

N2 - We present a generic immunoassay platform that uses enhanced total internal reflection fluorescence in the proximity of silver island films (SIFs), a surface coating consisting of metal (silver) particles. This platform is used with a model immunoassay where a protein antigen, rabbit immunoglobulin G, was immobilized on the SIF-coated glass surface. The signal from a fluorescent dye-labeled anti-rabbit antibody binding to the surface antigen was detected; different color dyes have been tested. Close placement of the fluorophore to surface-bound silver nanostructures results in dramatic signal enhancement (up to 40-fold) on the SIFs as compared with the glass slides. Use of the total internal reflection mode of excitation has significant advantages (over classic front-face excitation) for practical assay development. The limited evanescent wave excitation volume makes it possible to minimize the background signal and use the immunoassay with no need for any washing steps.

AB - We present a generic immunoassay platform that uses enhanced total internal reflection fluorescence in the proximity of silver island films (SIFs), a surface coating consisting of metal (silver) particles. This platform is used with a model immunoassay where a protein antigen, rabbit immunoglobulin G, was immobilized on the SIF-coated glass surface. The signal from a fluorescent dye-labeled anti-rabbit antibody binding to the surface antigen was detected; different color dyes have been tested. Close placement of the fluorophore to surface-bound silver nanostructures results in dramatic signal enhancement (up to 40-fold) on the SIFs as compared with the glass slides. Use of the total internal reflection mode of excitation has significant advantages (over classic front-face excitation) for practical assay development. The limited evanescent wave excitation volume makes it possible to minimize the background signal and use the immunoassay with no need for any washing steps.

KW - Enhanced fluorescence

KW - Fluorescence immunoassay

KW - Immunoglobulin

KW - Nano-size metallic particles

KW - Silver island films

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DO - 10.1016/j.ab.2004.08.034

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EP - 311

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces

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Keywords

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