Characterization and performance evaluation of in vivo and in vitro produced monoclonal anti-TNT antibodies for the detection of TNT

Charles, Paul T.; Shriver‐Lake, Lisa C.; Francesconi, Stephen C.; Churilla, Albert M.; Rangasammy, Jermain G.; Patterson, Charles H.; Deschamps, Jeffrey R.; Kusterbeck, Anne W.

doi:10.1016/j.jim.2003.09.010

Cited by 27 publications

(31 citation statements)

References 22 publications

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“…The use of this immunogenic approach leads one to believe the scFv binding region would have some recognition capabilities for both the di-and trinitroaromatic compounds. Overall, these crossreactivity results were in good agreement compared to previously reported antibody trials using monoclonal anti-TNT [32,33].…”

Section: Resultssupporting

confidence: 89%

Multi-channeled single chain variable fragment (scFv) based microfluidic device for explosives detection

Charles

Davis

Adams

et al. 2015

Talanta

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The development of explosives detection technologies has increased significantly over the years as environmental and national security agencies implement tighter pollution control measures and methods for improving homeland security. 2, 4, 6-Trinitrotoluene (TNT), known primarily as a component in munitions, has been targeted for both its toxicity and carcinogenic properties that if present at high concentrations can be a detriment to both humans, marine and plant ecosystems. Enabling end users with environmental detection and monitoring systems capable of providing real-time, qualitative and quantitative chemical analysis of these toxic compounds would be extremely beneficial. Reported herein is the development of a multi-channeled microfluidic device immobilized with single chain fragment variable (scFv) recombinant proteins specific for the explosive, TNT. Fluorescence displacement immunoassays performed under constant flow demonstrated trace level sensitivity and specificity for TNT. The utility of three multi-channeled devices immobilized with either (1) scFv recombinant protein, (2) biotinylated-scFv (bt-scFv) and (3) monoclonal anti-TNT (whole IgG molecule) were investigated and compared. Fluorescence dose response curves, crossreactivity measurements and limits of detection (LOD) for TNT were determined. Fluorescence displacement immunoassays for TNT in natural seawater demonstrated detection limits at sub-parts-per-billion levels (0.5 ppb) utilizing the microfluidic device with immobilized bt-scFv.

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Section: Resultssupporting

confidence: 89%

Multi-channeled single chain variable fragment (scFv) based microfluidic device for explosives detection

Charles

Davis

Adams

et al. 2015

Talanta

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“…The method worked well in tube and microtiter plate formats and achieved detection limits of 0.05 ppb for TNT in artificial sea water. Charles et al (2004a) characterized a number of monoclonal anti-TNT antibodies, and used them in a microcapillary displacement fluorescence immunoassay. This research group's flow Advances in Fluorescence Spectroscopy 857 micro-immunosensor method, using the cyanine dye Cy5 to label the antibodies, was described in detail in a second paper (Charles et al, 2004b).…”

Section: Fluorescence Detection Of Explosivesmentioning

confidence: 99%

Some Recent Advances in Fluorescence Spectroscopy

Miller

2006

Analytical Letters

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Fluorescence analysis methods continue to increase rapidly in importance in an ever-wider range of application areas. This growth highlights the need for the development and use of better standard materials and methods to ensure interlaboratory compatibility of results. Further impetus to the use of fluorescence methods has been provided by combining them with modern chemometric approaches that extract more information from multivariate fluorescence spectroscopy. An example of a new area where fluorescence methods, combined with biospecific and other sensors, show significant potential, is the detection of explosives' residues and components.

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“…Therefore, in response to growing concern about the health effects and the ecological threats posed by TNT, numerous recent studies have been focused on its toxicity (Dodard et al 2004;Patel et al 2004;Schaefer 2004), environmental fate (Lynch et al 2003;Kim et al 2004;Weiss et al 2004b, c), detection methods (Altamirano et al 2004;Borch & Gerlach 2004;Buryakov 2004;Charles et al 2004;Jung et al 2004;Weiss et al 2004a), size and mass distributions of TNT particles scattered by detonation (Taylor et al 2004) and, last but not least, (bio)degradation.…”

Section: Nitroaromatic Compoundsmentioning

confidence: 99%

Promising Strategies for the Mineralisation of 2,4,6-trinitrotoluene

Stenuit

Eyers

Fantroussi

et al. 2005

Rev Environ Sci Biotechnol

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2,4,6-trinitrotoluene (TNT) is known to be one of the most common military explosives. In spite of its established toxicity and mutagenicity for many organisms, soils and groundwater are still being frequently contaminated at manufacturing, disposal and TNT destruction sites. The inability of natural aquatic and soil biota to use TNT as growth substrate has been recognized as the primary limitation in the application of bioremediation processes to contaminated environments. However, promising degradation pathways have been recently discovered which may lead to the mineralisation of TNT. Significant advances have been made in studying the mechanism of TNT denitration, which can be considered as the major reaction and the driving force towards beneficial biodegradation. The possibilities to favour TNT denitration are discussed based on current knowledge of the enzymology and genetics of denitration in nitroaromatic degrading organisms. The literature survey demonstrates that the only enzymes characterized so far for their denitrase activity towards TNT belong to the class I flavin-dependent b/a barrel oxidoreductases, also known as the ''Old Yellow Enzyme'' family. In addition, this review provides an overview of strategies and future directions towards a rational search for new catabolic activities, including metagenomic library screening, plus new possibilities to improve the activity of known catabolic enzymes acting on TNT, such as DNA shuffling.

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Characterization and performance evaluation of in vivo and in vitro produced monoclonal anti-TNT antibodies for the detection of TNT

Cited by 27 publications

References 22 publications

Multi-channeled single chain variable fragment (scFv) based microfluidic device for explosives detection

Multi-channeled single chain variable fragment (scFv) based microfluidic device for explosives detection

Some Recent Advances in Fluorescence Spectroscopy

Promising Strategies for the Mineralisation of 2,4,6-trinitrotoluene

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