Porvided is a monoclonal antibody specific for hippuric acid which is one of the representative harmful haalucinogenic substances and is the major metabolites of toluene. In the present invention, a hippuric acid-carrier protein conjugate is prepared from hippuric 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.

The phosphate-specific transport substrate binding protein-1 (PstS1) is a potential antigen used for the serological diagnosis of tuberculosis. For a highly specific diagnostic result, it is important that the recombinant PstS1 be highly pure and correctly folded. In this study, the PstS1 was expressed as fusion protein with glutathione-S-transferase (PstS1-GST) and Escherichia coli trigger factor (PstS1-TF) and their immunodiagnostic potentials were evaluated. The insoluble PstS1-GST was denatured and refolded to the native conformation by a step-gradient dilution, followed by purification with affinity chromatography on immobilized glutathione whereas the soluble PstS1-TF was directly purified by Ni-NTA affinity and size-exclusion chromatographies.

This paper describes an electrochemical immunosensor for simple, fast and quantitative detection of a urinary hippuric acid which is one of major biological indicator in toluene-exposed humans. The feature of this electrochemical system for immunoassay of hippuric acid is based on the direct conjugation of ferrocene to a hippuric acid. With the competition between the ferrocene-hippuric acid complex and hippuric acid for binding to the anti-hippuric acid monoclonal antibody coated onto gold nanoparticles, the electrical signals are turned out to be proportional to urinary hippuric acid in the range of 0.01-10 mg/mL, which is enough to be used for the point-of-care.

It is not easy to produce the monoclonal antibody because of its restriction to mouse, labor and time consuming processes, and difficulties to regulate specificity and affinity for target antigen. The aims of this study are to product the recombinant antibody in E. coli and to establish the technical platforms to manipulate specificity and affinity of the recombinant antibody. DNA fragments for the variable regions of the heavy and the light chains were amplified by RT-PCR from the hybridoma cell producing the antibody against hippuric acid and joined by SOE-PCR with a linker DNA encoding a peptide (Gly4Ser)3, resulting in a single-chain variable fragment (scFv). The degenerated primers for PCR were designed and supposed to cover 78% and 75% of gamma(g) and kappa(k) genes in mouse, respectively.