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Issue 5, 2009
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An ultra-sensitive nanoarray chip based on single-molecule sandwich immunoassay and TIRFM for protein detection in biologic fluids

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Abstract

This paper describes a single-molecule sandwich immunoassay method that utilizes total internal reflection fluorescence microscopy (TIRFM) at the single-molecule level for nanoarray protein chip applications. Nanoarray patterning of a biotin-probe with a spot diameter of 179 ± 1 nm was performed successfully on a (3-mercaptopropyl)trimethoxysilane (MPTMS)-coated glass substrate by atomic force microscopy (AFM). The formation of biotin patterns was confirmed directly by observing the heights of bound streptavidin and biotin-antibody on glass substrates using an AFM in contact mode. Target protein molecules (or antigen) at the zepto-molar (zM) concentration level (× 10−21 M) were detected on MPTMS-coated glass nanoarray protein chips by TIRFM. Finally, cytokine clinical samples (i.e. TNF-α and IL-1α) as cancer marker protein molecules were applied to nanoarray protein chips, and detection limits were at 600 zM.

Graphical abstract: An ultra-sensitive nanoarray chip based on single-molecule sandwich immunoassay and TIRFM for protein detection in biologic fluids

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Publication details

The article was received on 09 Dec 2008, accepted on 17 Feb 2009 and first published on 02 Mar 2009


Article type: Paper
DOI: 10.1039/B822094H
Citation: Analyst, 2009,134, 933-938
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    An ultra-sensitive nanoarray chip based on single-molecule sandwich immunoassay and TIRFM for protein detection in biologic fluids

    S. Lee, N. Cho, J. D. Kim, H. Jung and S. H. Kang, Analyst, 2009, 134, 933
    DOI: 10.1039/B822094H

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