Homogeneous immunoassay for the detection of trinitrotoluene (TNT) based on the reactivation of apoglucose oxidase using a novel FAD-trinitrotoluene conjugate

M. Dosch; M. G. Weller; A. F. Bückmann; R. Niessner; A. F. Bückmann

doi:10.1007/s002160050857

Homogeneous immunoassay for the detection of trinitrotoluene (TNT) based on the reactivation of apoglucose oxidase using a novel FAD-trinitrotoluene conjugate

M. Dosch, M. G. Weller, A. F. Bückmann, R. Niessner, A. F. Bückmann

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

Abstract

A flavin adenine dinucleotide-trinitrotoluene derivative (FAD-TNT) was synthesized by coupling N⁶-(2-aminoethyl)-FAD covalently to the N-hydroxysuccinimidyl ester of trinitrophenyl-γ-aminobutyric acid and characterized by negative-ion electrospray-ionization mass spectrometry (ESI-MS) after purification by reversed-phase HPLC. Free FAD-TNT can be detected at very low levels by recombination with apoglucose oxidase, since the FAD-TNT-glucose oxidase complex is enzymatically active. On the contrary, if FAD-TNT has been bound by an anti-TNT antibody, the conjugate cannot recombine with apoglucose oxidase any more. Based on these two phenomena, a homogeneous apoenzyme reactivation immunoassay system (ARIS) was developed for the detection of TNT. No separation step is needed in this assay. Proportionality between the TNT concentration and enzyme activity was demonstrated with a detection limit of 5 μg/L TNT.

Original language	English
Pages (from-to)	174-178
Number of pages	5
Journal	Fresenius' Journal of Analytical Chemistry
Volume	361
Issue number	2
DOIs	https://doi.org/10.1007/s002160050857
State	Published - 1998
Externally published	Yes

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@article{b605cc8e6c9343ccb3a8ac2085bfa682,

title = "Homogeneous immunoassay for the detection of trinitrotoluene (TNT) based on the reactivation of apoglucose oxidase using a novel FAD-trinitrotoluene conjugate",

abstract = "A flavin adenine dinucleotide-trinitrotoluene derivative (FAD-TNT) was synthesized by coupling N6-(2-aminoethyl)-FAD covalently to the N-hydroxysuccinimidyl ester of trinitrophenyl-γ-aminobutyric acid and characterized by negative-ion electrospray-ionization mass spectrometry (ESI-MS) after purification by reversed-phase HPLC. Free FAD-TNT can be detected at very low levels by recombination with apoglucose oxidase, since the FAD-TNT-glucose oxidase complex is enzymatically active. On the contrary, if FAD-TNT has been bound by an anti-TNT antibody, the conjugate cannot recombine with apoglucose oxidase any more. Based on these two phenomena, a homogeneous apoenzyme reactivation immunoassay system (ARIS) was developed for the detection of TNT. No separation step is needed in this assay. Proportionality between the TNT concentration and enzyme activity was demonstrated with a detection limit of 5 μg/L TNT.",

author = "M. Dosch and Weller, {M. G.} and B{\"u}ckmann, {A. F.} and R. Niessner and B{\"u}ckmann, {A. F.}",

note = "Funding Information: Acknowledgements We would like to thank Strategic Diagnostic Inc., Newark, USA, for donating monoclonal antibody against TNT. In addition, financial support from Merck KGaA is gratefully acknowledged. Furthermore we kindly want to thank the Abteilung f{\"u}r Strukturaufkl{\"a}rung at the GBF, Braunschweig, for performing the ESI-MS.",

year = "1998",

doi = "10.1007/s002160050857",

language = "English",

volume = "361",

pages = "174--178",

journal = "Fresenius' Journal of Analytical Chemistry",

issn = "0937-0633",

publisher = "Springer Verlag",

number = "2",

}

TY - JOUR

T1 - Homogeneous immunoassay for the detection of trinitrotoluene (TNT) based on the reactivation of apoglucose oxidase using a novel FAD-trinitrotoluene conjugate

AU - Dosch, M.

AU - Weller, M. G.

AU - Bückmann, A. F.

AU - Niessner, R.

AU - Bückmann, A. F.

N1 - Funding Information: Acknowledgements We would like to thank Strategic Diagnostic Inc., Newark, USA, for donating monoclonal antibody against TNT. In addition, financial support from Merck KGaA is gratefully acknowledged. Furthermore we kindly want to thank the Abteilung für Strukturaufklärung at the GBF, Braunschweig, for performing the ESI-MS.

PY - 1998

Y1 - 1998

N2 - A flavin adenine dinucleotide-trinitrotoluene derivative (FAD-TNT) was synthesized by coupling N6-(2-aminoethyl)-FAD covalently to the N-hydroxysuccinimidyl ester of trinitrophenyl-γ-aminobutyric acid and characterized by negative-ion electrospray-ionization mass spectrometry (ESI-MS) after purification by reversed-phase HPLC. Free FAD-TNT can be detected at very low levels by recombination with apoglucose oxidase, since the FAD-TNT-glucose oxidase complex is enzymatically active. On the contrary, if FAD-TNT has been bound by an anti-TNT antibody, the conjugate cannot recombine with apoglucose oxidase any more. Based on these two phenomena, a homogeneous apoenzyme reactivation immunoassay system (ARIS) was developed for the detection of TNT. No separation step is needed in this assay. Proportionality between the TNT concentration and enzyme activity was demonstrated with a detection limit of 5 μg/L TNT.

AB - A flavin adenine dinucleotide-trinitrotoluene derivative (FAD-TNT) was synthesized by coupling N6-(2-aminoethyl)-FAD covalently to the N-hydroxysuccinimidyl ester of trinitrophenyl-γ-aminobutyric acid and characterized by negative-ion electrospray-ionization mass spectrometry (ESI-MS) after purification by reversed-phase HPLC. Free FAD-TNT can be detected at very low levels by recombination with apoglucose oxidase, since the FAD-TNT-glucose oxidase complex is enzymatically active. On the contrary, if FAD-TNT has been bound by an anti-TNT antibody, the conjugate cannot recombine with apoglucose oxidase any more. Based on these two phenomena, a homogeneous apoenzyme reactivation immunoassay system (ARIS) was developed for the detection of TNT. No separation step is needed in this assay. Proportionality between the TNT concentration and enzyme activity was demonstrated with a detection limit of 5 μg/L TNT.

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DO - 10.1007/s002160050857

M3 - Article

AN - SCOPUS:0000877997

SN - 0937-0633

VL - 361

SP - 174

EP - 178

JO - Fresenius' Journal of Analytical Chemistry

JF - Fresenius' Journal of Analytical Chemistry

IS - 2

ER -

Homogeneous immunoassay for the detection of trinitrotoluene (TNT) based on the reactivation of apoglucose oxidase using a novel FAD-trinitrotoluene conjugate

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