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 language | English |
|---|---|
| Pages (from-to) | 933-938 |
| Number of pages | 6 |
| Journal | Analyst |
| Volume | 134 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2009 May 5 |
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All Science Journal Classification (ASJC) codes
- Analytical Chemistry
- Biochemistry
- Environmental Chemistry
- Spectroscopy
- Electrochemistry
Cite this
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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 journal › Article
TY - JOUR
T1 - An ultra-sensitive nanoarray chip based on single-molecule sandwich immunoassay and TIRFM for protein detection in biologic fluids
AU - Lee, Seungah
AU - Cho, Nam Pyo
AU - Kim, Jung Dong
AU - Jung, Hyungil
AU - Kang, Seong Ho
PY - 2009/5/5
Y1 - 2009/5/5
N2 - 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.
AB - 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.
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U2 - 10.1039/b822094h
DO - 10.1039/b822094h
M3 - Article
VL - 134
SP - 933
EP - 938
JO - The Analyst
JF - The Analyst
SN - 0003-2654
IS - 5
ER -