title: Multi-channeled single chain variable fragment (scFv) based microfluidic device for explosives detection.
abstract: 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.
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA. Electronic address: paul.charles@nrl.navy.mil.
authors.forename: Paul T
authors.initials: PT
authors.lastname: Charles
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
authors.forename: Jasmine
authors.initials: J
authors.lastname: Davis
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
authors.forename: André A
authors.initials: AA
authors.lastname: Adams
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
authors.forename: George P
authors.initials: GP
authors.lastname: Anderson
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
authors.forename: Jinny L
authors.initials: JL
authors.lastname: Liu
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Deschamps
authors.affiliation: Center for Bio/Molecular Science and Engineering (Code 6920), US Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
authors.forename: Anne W
authors.initials: AW
authors.lastname: Kusterbeck
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001360:Avidin / Q000737:chemistry; D019921:Biotinylation; D053834:Explosive Agents / Q000032:analysis* / Q000276:immunology; D005453:Fluorescence; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D012623:Seawater / Q000737:chemistry; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology*; D014303:Trinitrotoluene / Q000032:analysis* / Q000276:immunology; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000276:immunology
chemical_list: D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D053834:Explosive Agents; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014874:Water Pollutants, Chemical; C093483:neutravidin; D014303:Trinitrotoluene; D001360:Avidin
keywords: Fluorescence; Immunosensor; Microchannel; Recombinant protein; TNT; scFv