Abstract: Provided is a monoclonal antibody specific for hippuric acid. In the present invention, a hippuric acid-carrier protein conjugate is prepared from hippuric acid and BSA or OVA as a carrier protein, using a coupling reagent and a cross-linker. The monoclonal antibody screened according to the present invention has a titer having a standard curve in the concentration range of mg/mL meeting requirements for the permissible exposure limit (PEL) of toluene. The monoclonal antibody exhibits no cross-reactivity with carrier proteins, exhibits higher competitive inhibition in response to an increasing concentration of hippuric acid, and exhibits no cross-reactivity with other proteins contained in the urine. Therefore, the disclosed monoclonal antibody can be usefully employed in a diagnostic kit for detection of hippuric acid which is capable of diagnosing toluene exposure.

Abstract: Provided is a monoclonal antibody specific for hippuric acid which is one of the representative harmful hallucinogenic substances and is the major metabolite of toluene. In the present invention, a hippuric acid-carrier protein conjugate is prepared from hippuric acid and BSA or OVA as a carrier protein, using a coupling reagent and a cross-linker, mice are immunized by injection of the resulting conjugate, splenocytes are collected from animals and fused with myeloma cells, fused cells are cultured in HAT medium, a cell line producing a hippuric acid-directed monoclonal antibody is screened using a detection conjugate, a gene coding for variable regions of the screened monoclonal antibody is amplified by RT-PCR and then amplified by SOE-PCR using a linker DNA to thereby prepare a single chain variable fragment (scFV) which is cloned into a vector, thereby sequencing the base sequence and deducing amino acid sequences of the variable regions.