PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST CLASS-THETA GLUTATHIONE-S-TRANSFERASE T1-1

The recently discovered human class theta glutathione S-transferase T1 -1 (GSTT1-1) is responsible for the GSH-dependent detoxification of na turally occurring monohalomethanes. The detoxifying role of GSTT1-1 ha s not been investigated in cancer susceptibility and the polymorphism of the protein is unknown in different populations. The purpose of our work was to produce a panel of mouse monoclonal antibodies (MAbs) tha t could bind to different regions of the GSTT1-1 protein and would hel p us select suitable MAbs for Western blot analyses and immunohistoche mistry, and develop an ELISA assay for detection of GSTT1-1 in whole b lood, Six highly specific MAbs were generated against GSTT1-1. Out of six MAbs, one was able to recognize only the native form of the enzyme and possesses two binding sites on the dimeric GSTT1-1 molecule. The other five MAbs bind to both native and denatured GSTT1-1 enzyme in di rect and antigen capture ELISA or Western blot. The antibodies recogni ze at least four different epitopes on the GSTT1-1 molecule, Using MAb s 4G1 and 2D8, a sensitive ELISA assay for determination of GSTT1-1 in whole blood was developed.