2,4,6-Trinitrotoluene detection using recombinant antibodies.

The ability to detect low levels of 2,4,6-trinitrotoluene (TNT) in aqueous samples is important due to the toxicity of both TNT and its breakdown products. We have been characterizing recombinant anti-TNT antibodies isolated from the Griffin library of phage displayed scFvs by selection for binders to the TNT-surrogate 2,4,6-trinitrobenzene (TNB) coupled to the protein bovine serum albumin. Two candidate antibody fragments, TNB1 and TNB2, were isolated and evaluated by ELISA for their ability to bind to TNB coupled to the protein ovalbumin. Competition ELISA was then used to demonstrate antibody fragment binding to TNT in solution and to examine cross-reactivity towards several TNT-related compounds and other explosives. Both recombinant antibody fragments were incorporated into a continuous flow assay for the detection of TNT. TNB2, the best single chain antibody, showed a limit of detection of 1 ng ml(-1), comparable to a commercially available anti-TNT antibody in the same assay format.