Preparation of Anti-2,4-dinitrotoluene Monoclonal Antibody by Using Rat Medial Iliac Lymph Node Cells and Its Characterization Using Solid Phase Enzyme-linked Immunosorbent Assay

Recent concern on international terrorism and weapons of mass destruction demands the development of novel analytical methods for identification and quantification of explosive molecules. 2,4-Dinitrotoluene (2,4-DNT) is one of the decomposed compounds from 2,4,6-trinitrotoluene (TNT), a main explosive compound, and has high vapor pressure and life time than TNT. 2,4-DNT, thus, become a chemical signature of explosives. Monoclonal antibody against 2,4-DNT was prepared by using rat lymph node method. Four hybridoma cells which produce anti-DNPh antibody were produced out of 61 fusion positive cells. The best hybridoma cell (named DNPh-mAb2 cell) was selected for the production of monoclonal antibody (DNPh-mAb2). The properties of DNPh-mAb2 were investigated such as cross-reactivity and affinity constant. The prepared DNPh-mAb2 has high avidity to 2,4-DNT compared with other nitroaromatic compounds except N- (2,4,6-tirinitrophenyl) glycine (TNP-Gly, ca. 50%). TNP-Gly is not contained in general explosives, appearing no problem. The antibody has high affinity constant of 3.610 -6 mol L -1 to 2,4-DNT. The limit of detection of 2,4-DNT by indirect competitive ELISA was ca. 5 ng mL -1 , showing suitable for 2,4-DNT detection.