Preparation of a Polyclonal Antibody and a Bioassay for Nitroaromatic Compounds by an Enzyme-Linked Immunosorbent Assay Technique and a Surface Plasmon Resonance Biosensor

Takatoshi Sakai, Akira Torimaru, Katsura Shinahara, Norio Miura, Toshihiko Imato, Kiyoshi Toko, Kiyoshi Matsumoto

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Sensitive and selective detection of 2,4,6-trinitrophenyl derivatives (TNP derivatives) based on surface plasmon resonance (SPR) was performed using polyclonal anti-TNP antibody and N-(TNP)-ovalbumin (TNP-OVA) conjugate. TNP-bovine serum albumin (TNP-BSA) conjugate was injected into the mice, and polyclonal anti-TNP antibody was gained after purification of the sera using protein G. TNP-OVA was immobilized on the Au film of the SPR sensor chip by physical adsorption. The incidence angle shift of the TNP-OVA immobilized sensor increased steeply with increasing concentration of antiTNP antibody up to 20 μg/ml and increased only slightly above this concentration. The additions of TNP derivatives into the anti-TNP antibody solution were found to decrease the incidence angle shift because of the inhibition effects of TNP derivatives. The lowest detection limit for trinitrotoluene (TNT) by SPR was 1×10-7 g/ml, whereas for TNP-6-aminohexanoic acid (TNP-aha) it was 3×10-9 g/ml. Evaluations of affinity constants of anti-TNP antibody were performed. Analyses of SPR data were carried out by assumptions of the Langmuir isotherm and equilibrium state of immunoreaction. The value of association constant between anti-TNP antibody and immobilized TNP-OVA (K 1) was 6.4×106 M-1. The values of association constants between antibody and TNP-aha (K2) were 2.7 and 8.5×106 M-1 when 20 and 10 μg/ml antibody were used, respectively.

Original languageEnglish
Pages (from-to)439-452
Number of pages14
JournalSensors and Materials
Volume15
Issue number8
Publication statusPublished - Dec 1 2003

Fingerprint

Immunosorbents
bioassay
Bioassay
Surface plasmon resonance
antibodies
bioinstrumentation
surface plasmon resonance
Biosensors
Antibodies
enzymes
Assays
Enzymes
preparation
Aminocaproic Acid
Derivatives
serums
Association reactions
Trinitrotoluene
Immobilized Antibodies
incidence

All Science Journal Classification (ASJC) codes

  • Instrumentation
  • Materials Science(all)

Cite this

Preparation of a Polyclonal Antibody and a Bioassay for Nitroaromatic Compounds by an Enzyme-Linked Immunosorbent Assay Technique and a Surface Plasmon Resonance Biosensor. / Sakai, Takatoshi; Torimaru, Akira; Shinahara, Katsura; Miura, Norio; Imato, Toshihiko; Toko, Kiyoshi; Matsumoto, Kiyoshi.

In: Sensors and Materials, Vol. 15, No. 8, 01.12.2003, p. 439-452.

Research output: Contribution to journalArticle

Sakai, Takatoshi ; Torimaru, Akira ; Shinahara, Katsura ; Miura, Norio ; Imato, Toshihiko ; Toko, Kiyoshi ; Matsumoto, Kiyoshi. / Preparation of a Polyclonal Antibody and a Bioassay for Nitroaromatic Compounds by an Enzyme-Linked Immunosorbent Assay Technique and a Surface Plasmon Resonance Biosensor. In: Sensors and Materials. 2003 ; Vol. 15, No. 8. pp. 439-452.
@article{99a1cd11c7634d6d8eea6693d767a65c,
title = "Preparation of a Polyclonal Antibody and a Bioassay for Nitroaromatic Compounds by an Enzyme-Linked Immunosorbent Assay Technique and a Surface Plasmon Resonance Biosensor",
abstract = "Sensitive and selective detection of 2,4,6-trinitrophenyl derivatives (TNP derivatives) based on surface plasmon resonance (SPR) was performed using polyclonal anti-TNP antibody and N-(TNP)-ovalbumin (TNP-OVA) conjugate. TNP-bovine serum albumin (TNP-BSA) conjugate was injected into the mice, and polyclonal anti-TNP antibody was gained after purification of the sera using protein G. TNP-OVA was immobilized on the Au film of the SPR sensor chip by physical adsorption. The incidence angle shift of the TNP-OVA immobilized sensor increased steeply with increasing concentration of antiTNP antibody up to 20 μg/ml and increased only slightly above this concentration. The additions of TNP derivatives into the anti-TNP antibody solution were found to decrease the incidence angle shift because of the inhibition effects of TNP derivatives. The lowest detection limit for trinitrotoluene (TNT) by SPR was 1×10-7 g/ml, whereas for TNP-6-aminohexanoic acid (TNP-aha) it was 3×10-9 g/ml. Evaluations of affinity constants of anti-TNP antibody were performed. Analyses of SPR data were carried out by assumptions of the Langmuir isotherm and equilibrium state of immunoreaction. The value of association constant between anti-TNP antibody and immobilized TNP-OVA (K 1) was 6.4×106 M-1. The values of association constants between antibody and TNP-aha (K2) were 2.7 and 8.5×106 M-1 when 20 and 10 μg/ml antibody were used, respectively.",
author = "Takatoshi Sakai and Akira Torimaru and Katsura Shinahara and Norio Miura and Toshihiko Imato and Kiyoshi Toko and Kiyoshi Matsumoto",
year = "2003",
month = "12",
day = "1",
language = "English",
volume = "15",
pages = "439--452",
journal = "Sensors and Materials",
issn = "0914-4935",
publisher = "M Y U Scientific Publishing Division",
number = "8",

}

TY - JOUR

T1 - Preparation of a Polyclonal Antibody and a Bioassay for Nitroaromatic Compounds by an Enzyme-Linked Immunosorbent Assay Technique and a Surface Plasmon Resonance Biosensor

AU - Sakai, Takatoshi

AU - Torimaru, Akira

AU - Shinahara, Katsura

AU - Miura, Norio

AU - Imato, Toshihiko

AU - Toko, Kiyoshi

AU - Matsumoto, Kiyoshi

PY - 2003/12/1

Y1 - 2003/12/1

N2 - Sensitive and selective detection of 2,4,6-trinitrophenyl derivatives (TNP derivatives) based on surface plasmon resonance (SPR) was performed using polyclonal anti-TNP antibody and N-(TNP)-ovalbumin (TNP-OVA) conjugate. TNP-bovine serum albumin (TNP-BSA) conjugate was injected into the mice, and polyclonal anti-TNP antibody was gained after purification of the sera using protein G. TNP-OVA was immobilized on the Au film of the SPR sensor chip by physical adsorption. The incidence angle shift of the TNP-OVA immobilized sensor increased steeply with increasing concentration of antiTNP antibody up to 20 μg/ml and increased only slightly above this concentration. The additions of TNP derivatives into the anti-TNP antibody solution were found to decrease the incidence angle shift because of the inhibition effects of TNP derivatives. The lowest detection limit for trinitrotoluene (TNT) by SPR was 1×10-7 g/ml, whereas for TNP-6-aminohexanoic acid (TNP-aha) it was 3×10-9 g/ml. Evaluations of affinity constants of anti-TNP antibody were performed. Analyses of SPR data were carried out by assumptions of the Langmuir isotherm and equilibrium state of immunoreaction. The value of association constant between anti-TNP antibody and immobilized TNP-OVA (K 1) was 6.4×106 M-1. The values of association constants between antibody and TNP-aha (K2) were 2.7 and 8.5×106 M-1 when 20 and 10 μg/ml antibody were used, respectively.

AB - Sensitive and selective detection of 2,4,6-trinitrophenyl derivatives (TNP derivatives) based on surface plasmon resonance (SPR) was performed using polyclonal anti-TNP antibody and N-(TNP)-ovalbumin (TNP-OVA) conjugate. TNP-bovine serum albumin (TNP-BSA) conjugate was injected into the mice, and polyclonal anti-TNP antibody was gained after purification of the sera using protein G. TNP-OVA was immobilized on the Au film of the SPR sensor chip by physical adsorption. The incidence angle shift of the TNP-OVA immobilized sensor increased steeply with increasing concentration of antiTNP antibody up to 20 μg/ml and increased only slightly above this concentration. The additions of TNP derivatives into the anti-TNP antibody solution were found to decrease the incidence angle shift because of the inhibition effects of TNP derivatives. The lowest detection limit for trinitrotoluene (TNT) by SPR was 1×10-7 g/ml, whereas for TNP-6-aminohexanoic acid (TNP-aha) it was 3×10-9 g/ml. Evaluations of affinity constants of anti-TNP antibody were performed. Analyses of SPR data were carried out by assumptions of the Langmuir isotherm and equilibrium state of immunoreaction. The value of association constant between anti-TNP antibody and immobilized TNP-OVA (K 1) was 6.4×106 M-1. The values of association constants between antibody and TNP-aha (K2) were 2.7 and 8.5×106 M-1 when 20 and 10 μg/ml antibody were used, respectively.

UR - http://www.scopus.com/inward/record.url?scp=1542501616&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1542501616&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:1542501616

VL - 15

SP - 439

EP - 452

JO - Sensors and Materials

JF - Sensors and Materials

SN - 0914-4935

IS - 8

ER -