Characterization and performance evaluation of in vivo and in vitro produced monoclonal anti-TNT antibodies for the detection of TNT.
In this paper are the experimental results used to characterize four distinct monoclonal anti-TNT antibodies (in vivo and in vitro cloned) for potential use in a field-portable immunosensor. Direct and competitive enzyme-linked immunosorbent assays (ELISA) were performed to determine their affinity for TNT and a fluorescently labeled analog of TNT for use in an immunosensor. Effective concentrations (EC(50)), inhibition concentration (IC(50)) and cross-reactivity measurements to related nitroaromatics (e.g., 2,4,6-trinitrobenzene [TNB], methyl-2,4,6-trinitrophenyl nitramine [tetryl], 2-amino-4,6-dinitrotoluene [2A-4,6-DNT], 2,4-dinitrotoluene [2,4-DNT] and 1,3-dinitrotoluene [1,3-DNT]) were measured. Final characterization of the monoclonal antibodies was based on performance (measured by fluorescence dose response) using a fluorescence-based microcapillary displacement assay. Analytical techniques showed a high degree of affinity for TNT and varying degrees of cross-reactivity with each respective monoclonal antibody. Microcapillary displacement immunoassays with each of the antibodies resulted in detection capabilities at the lowest applied TNT concentration (10 ng/ml).
Charles
Paul T
PT
The Center for Bio/Molecular Science and Engineering Code 6900, The U.S. Naval Research Laboratory, 4555 Overlook Ave, SW, Washington, DC 20375, USA. ptc@cbmse.nrl.navy.mil
Shriver-Lake
Lisa C
LC
Francesconi
Stephen C
SC
Churilla
Albert M
AM
Rangasammy
Jermain G
JG
Patterson
Charles H
CH
Deschamps
Jeffrey R
JR
Kusterbeck
Anne W
AW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D003429:Cross Reactions; D004784:Environmental Monitoring / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D012989:Soil Pollutants / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D014303:Trinitrotoluene / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D012989:Soil Pollutants; D014303:Trinitrotoluene