Multianalyte detection using a capillary-based flow immunosensor

Abstract

A highly sensitive, dual-analyte detection system using capillary-based immunosensors has been designed for explosive detection. This model system consists of two capillaries, one coated with antibodies specific for 2,4,6- trinitrotoluene (TNT) and the other specific for hexahydro-1,3,5-trinitro- 1,3,5-triazine (RDX) combined into a single device. The fused silica capillaries are prepared by coating anti-TNT and anti-RDX antibodies onto the silanized inner walls using a hetero-bifunctional crosslinker. After immobilization, the antibodies are saturated with a suitable fluorophore- labeled antigen. A 'T' connector is used to continuously flow the buffer solution through the individual capillaries. To perform the assay, an aliquot of TNT or RDX or a mixture of the two analyses is injected into the continuous flow stream. In each capillary, the target analyte displaces the fluorophore-labeled antigen from the binding pocket of the antibody. The labeled antigen displaced from either capillary is detected downstream using two portable spectrofluorometers. The limits of detection for TNT and RDX in the multianalyte format are 44 fmol (100 μl of 0.1 ng/ml TNT solution) and 224 fmol (100 μl of 0.5 ng/ml RDX solution), respectively. The entire assay for both analytes can be performed in less than 3 min.