Development and comparison of two immunoassays for the detection of 2,4,6-trinitrotoluene (TNT) based on surface plasmon resonance

Dhesingh Ravi Shankaran, Kiyoshi Matsumoto, Kiyoshi Toko, Norio Miura

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

This paper describes the development and comparison of two immunoassays for the detection of 2,4,6-trinitrotoluene (TNT) based on competitive inhibition. Two polyclonal antibodies were used in the immunoassay; one was prepared from 2,4,6-trinitrophenol-bovine serum albumin conjugate (anti-TNP-BSA Ab, goat IgG) and another from 2,4,6-trinitrophenyl-keyhole limpet hemocyanine conjugate (anti-TNPh-KLH Ab, rabbit IgG). A solid-phase immunoassay that consisted of physically immobilized TNP-BSA conjugate was allowed to interact with these antibodies and the resonance angle changes due to the biomolecular interactions were monitored by surface plasmon resonance (SPR). The quantification of TNT was based on the change in the resonance angle shift for binding of antibodies with a TNP-BSA conjugate, which occurred in the presence of TNT in solution. Both antibodies showed a high degree of affinity for TNT and varying degrees of cross-reactivity to the related nitroaromatic derivative compounds, such as 2,4-dinitrotoluene (DNT), 1,3-dinitrobenzene (DNB), 2-amino-4,6-dintitrotoluene (2A-4,6-DNT), 4-amino-2,6-dinitrotoluene (4A-2,6-DNT) and TNP. The anti-TNPh-KLH Ab prepared in our laboratory showed a detection limit of 0.006 ng/ml (6 ppt), which was comparable to or even better than the detection limit (0.095 ng/ml or 95 ppt) of a commercially available anti-TNP-BSA Ab for detection of TNT. A pepsin solution prepared with a glycine-HCl buffer (pH 2.0) was used for regeneration of the sensor surface. The duration for a single immunocycle was ∼19 and ∼13 min for anti-TNP-BSA Ab and anti-TNPh-KLH Ab immunosystems, respectively. The proposed immunoassay is simple and possesses good analytical characteristics for quantification of TNT.

Original languageEnglish
Pages (from-to)71-79
Number of pages9
JournalSensors and Actuators, B: Chemical
Volume114
Issue number1
DOIs
Publication statusPublished - Mar 30 2006

Fingerprint

Trinitrotoluene
immunoassay
trinitrotoluene
2,4-dinitrotoluene
Surface plasmon resonance
surface plasmon resonance
Antibodies
antibodies
Immunoglobulin G
pepsin
goats
Amino acids
Pepsin A
rabbits
Bovine Serum Albumin
glycine
Derivatives
albumins
regeneration
serums

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Instrumentation
  • Condensed Matter Physics
  • Surfaces, Coatings and Films
  • Metals and Alloys
  • Electrical and Electronic Engineering
  • Materials Chemistry

Cite this

Development and comparison of two immunoassays for the detection of 2,4,6-trinitrotoluene (TNT) based on surface plasmon resonance. / Shankaran, Dhesingh Ravi; Matsumoto, Kiyoshi; Toko, Kiyoshi; Miura, Norio.

In: Sensors and Actuators, B: Chemical, Vol. 114, No. 1, 30.03.2006, p. 71-79.

Research output: Contribution to journalArticle

Shankaran, Dhesingh Ravi ; Matsumoto, Kiyoshi ; Toko, Kiyoshi ; Miura, Norio. / Development and comparison of two immunoassays for the detection of 2,4,6-trinitrotoluene (TNT) based on surface plasmon resonance. In: Sensors and Actuators, B: Chemical. 2006 ; Vol. 114, No. 1. pp. 71-79.
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