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

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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 3.6×10^-6mol 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.
2009-02-27