Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance

Toshikazu Kawaguchi, Dhesingh Ravi Shankaran, S. J. Kim, K. Vengatajalabathy Gobi, Kiyoshi Matsumoto, Kiyoshi Toko, Norio Miura

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

We have developed a new immunosensor based on self-assembly chemistry for highly sensitive and label-free detection of 2,4,6-trinitrotoluene (TNT) using surface plasmon resonance (SPR). A monolayer of amine terminated poly(ethylene glycol) hydrazinehydrochloride (PEG-NH2) thiolate was constructed on an activated gold surface and immobilized with trinitrophenyl-β-alanine (TNPh-β-alanine) by amide coupling method. The binding interaction of a monoclonal anti-TNT Ab (M-TNT Ab) with TNPh-β-alanine immobilized thiolate monolayer surface was monitored and evaluated for detection of TNT based on the principle of indirect competitive immunoreaction. Here, the competition between the self-assembled TNT derivative and the TNT in solution for binding with antibody yields in the response signal that is inversely proportional to the concentration of TNT in the linear detection range. With the present immunoassay format, TNT could be detected in the concentration range from 0.008 ng/ml (8 ppt) to 30 ng/ml (30 ppb). The response time for an immunoreaction was 2 min and one immunocycle could be done with in 4 min including surface regeneration. Bound antibodies could be easily eluted from the self-assembled immunosurface at high recoveries (more than 100 cycles) using pepsin solution without any damage to the TNT derivatives immobilized on the surface. The compact self-assembled monolayer was highly stable and prevented the non-specific adsorption of proteins on the surface favoring error free measurement.

Original languageEnglish
Pages (from-to)554-560
Number of pages7
JournalTalanta
Volume72
Issue number2
DOIs
Publication statusPublished - Apr 30 2007

Fingerprint

Trinitrotoluene
Immunosensors
Surface Plasmon Resonance
Surface plasmon resonance
Self assembled monolayers
Fabrication
Alanine
Polyethylene glycols
Monolayers
Derivatives
Ethylene Glycol
Antibodies
Pepsin A
Immunoassay
Amides
Gold
Self assembly
Adsorption
Reaction Time
Amines

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance. / Kawaguchi, Toshikazu; Shankaran, Dhesingh Ravi; Kim, S. J.; Gobi, K. Vengatajalabathy; Matsumoto, Kiyoshi; Toko, Kiyoshi; Miura, Norio.

In: Talanta, Vol. 72, No. 2, 30.04.2007, p. 554-560.

Research output: Contribution to journalArticle

Kawaguchi, Toshikazu ; Shankaran, Dhesingh Ravi ; Kim, S. J. ; Gobi, K. Vengatajalabathy ; Matsumoto, Kiyoshi ; Toko, Kiyoshi ; Miura, Norio. / Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance. In: Talanta. 2007 ; Vol. 72, No. 2. pp. 554-560.
@article{bf4304e8a9e0407e96e0ec04ddcdd8f0,
title = "Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance",
abstract = "We have developed a new immunosensor based on self-assembly chemistry for highly sensitive and label-free detection of 2,4,6-trinitrotoluene (TNT) using surface plasmon resonance (SPR). A monolayer of amine terminated poly(ethylene glycol) hydrazinehydrochloride (PEG-NH2) thiolate was constructed on an activated gold surface and immobilized with trinitrophenyl-β-alanine (TNPh-β-alanine) by amide coupling method. The binding interaction of a monoclonal anti-TNT Ab (M-TNT Ab) with TNPh-β-alanine immobilized thiolate monolayer surface was monitored and evaluated for detection of TNT based on the principle of indirect competitive immunoreaction. Here, the competition between the self-assembled TNT derivative and the TNT in solution for binding with antibody yields in the response signal that is inversely proportional to the concentration of TNT in the linear detection range. With the present immunoassay format, TNT could be detected in the concentration range from 0.008 ng/ml (8 ppt) to 30 ng/ml (30 ppb). The response time for an immunoreaction was 2 min and one immunocycle could be done with in 4 min including surface regeneration. Bound antibodies could be easily eluted from the self-assembled immunosurface at high recoveries (more than 100 cycles) using pepsin solution without any damage to the TNT derivatives immobilized on the surface. The compact self-assembled monolayer was highly stable and prevented the non-specific adsorption of proteins on the surface favoring error free measurement.",
author = "Toshikazu Kawaguchi and Shankaran, {Dhesingh Ravi} and Kim, {S. J.} and Gobi, {K. Vengatajalabathy} and Kiyoshi Matsumoto and Kiyoshi Toko and Norio Miura",
year = "2007",
month = "4",
day = "30",
doi = "10.1016/j.talanta.2006.11.020",
language = "English",
volume = "72",
pages = "554--560",
journal = "Talanta",
issn = "0039-9140",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance

AU - Kawaguchi, Toshikazu

AU - Shankaran, Dhesingh Ravi

AU - Kim, S. J.

AU - Gobi, K. Vengatajalabathy

AU - Matsumoto, Kiyoshi

AU - Toko, Kiyoshi

AU - Miura, Norio

PY - 2007/4/30

Y1 - 2007/4/30

N2 - We have developed a new immunosensor based on self-assembly chemistry for highly sensitive and label-free detection of 2,4,6-trinitrotoluene (TNT) using surface plasmon resonance (SPR). A monolayer of amine terminated poly(ethylene glycol) hydrazinehydrochloride (PEG-NH2) thiolate was constructed on an activated gold surface and immobilized with trinitrophenyl-β-alanine (TNPh-β-alanine) by amide coupling method. The binding interaction of a monoclonal anti-TNT Ab (M-TNT Ab) with TNPh-β-alanine immobilized thiolate monolayer surface was monitored and evaluated for detection of TNT based on the principle of indirect competitive immunoreaction. Here, the competition between the self-assembled TNT derivative and the TNT in solution for binding with antibody yields in the response signal that is inversely proportional to the concentration of TNT in the linear detection range. With the present immunoassay format, TNT could be detected in the concentration range from 0.008 ng/ml (8 ppt) to 30 ng/ml (30 ppb). The response time for an immunoreaction was 2 min and one immunocycle could be done with in 4 min including surface regeneration. Bound antibodies could be easily eluted from the self-assembled immunosurface at high recoveries (more than 100 cycles) using pepsin solution without any damage to the TNT derivatives immobilized on the surface. The compact self-assembled monolayer was highly stable and prevented the non-specific adsorption of proteins on the surface favoring error free measurement.

AB - We have developed a new immunosensor based on self-assembly chemistry for highly sensitive and label-free detection of 2,4,6-trinitrotoluene (TNT) using surface plasmon resonance (SPR). A monolayer of amine terminated poly(ethylene glycol) hydrazinehydrochloride (PEG-NH2) thiolate was constructed on an activated gold surface and immobilized with trinitrophenyl-β-alanine (TNPh-β-alanine) by amide coupling method. The binding interaction of a monoclonal anti-TNT Ab (M-TNT Ab) with TNPh-β-alanine immobilized thiolate monolayer surface was monitored and evaluated for detection of TNT based on the principle of indirect competitive immunoreaction. Here, the competition between the self-assembled TNT derivative and the TNT in solution for binding with antibody yields in the response signal that is inversely proportional to the concentration of TNT in the linear detection range. With the present immunoassay format, TNT could be detected in the concentration range from 0.008 ng/ml (8 ppt) to 30 ng/ml (30 ppb). The response time for an immunoreaction was 2 min and one immunocycle could be done with in 4 min including surface regeneration. Bound antibodies could be easily eluted from the self-assembled immunosurface at high recoveries (more than 100 cycles) using pepsin solution without any damage to the TNT derivatives immobilized on the surface. The compact self-assembled monolayer was highly stable and prevented the non-specific adsorption of proteins on the surface favoring error free measurement.

UR - http://www.scopus.com/inward/record.url?scp=34047257975&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34047257975&partnerID=8YFLogxK

U2 - 10.1016/j.talanta.2006.11.020

DO - 10.1016/j.talanta.2006.11.020

M3 - Article

C2 - 19071654

AN - SCOPUS:34047257975

VL - 72

SP - 554

EP - 560

JO - Talanta

JF - Talanta

SN - 0039-9140

IS - 2

ER -