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

Seungah Lee, Nam Pyo Cho, Jung Dong Kim, Hyungil Jung, Seong Ho Kang

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)933-938
Number of pages6
JournalAnalyst
Volume134
Issue number5
DOIs
Publication statusPublished - 2009 May 5

Fingerprint

Protein Array Analysis
Fluorescence microscopy
immunoassay
Biotin
Fluorescence Microscopy
Immunoassay
Glass
microscopy
fluorescence
Atomic Force Microscopy
Proteins
Molecules
protein
Fluids
fluid
atomic force microscopy
glass
Streptavidin
Atomic force microscopy
Interleukin-1

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

Lee, Seungah ; Cho, Nam Pyo ; Kim, Jung Dong ; Jung, Hyungil ; Kang, Seong Ho. / An ultra-sensitive nanoarray chip based on single-molecule sandwich immunoassay and TIRFM for protein detection in biologic fluids. In: Analyst. 2009 ; Vol. 134, No. 5. pp. 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. / Lee, Seungah; Cho, Nam Pyo; Kim, Jung Dong; Jung, Hyungil; Kang, Seong Ho.

In: Analyst, Vol. 134, No. 5, 05.05.2009, p. 933-938.

Research output: Contribution to journalArticle

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