Isolation of anti-glutathione antibodies from a phage display library

We have isolated anti-glutathione antibodies from a human synthetic phageantibody scFv library (Nissim,A., Hoogenboom,H.R., Tomlinson,I.M.,Flynn,G., Midgley,C., Lane,D. and Winter,G., 1994, EMBO J., 13, 692-698).Glutathione (GSH) conjugates with carrier proteins, such as bovine serumalbumin (BSA), keyhole limpet hemocyanin (KLH) and human lysozyme (LZM),were used as antigens. After four cycles of panning and affinitychromatography, clones that recognized GSH- conjugated proteins, but notBSA, KLH or LZM, were isolated. The isolated phage antibodies and thesoluble scFv fragments were characterized by immunoblotting, and thenucleotide sequences of the VH segments of selected clones were determined.The binding of several isolates to GSH-BSA was competitively inhibited byGSH in an ELISA. These observations have demonstrated that antibodiesagainst GSH, a tripeptide, can be isolated from the library. We constructedthe tertiary models of several scFv fragments and discussed the mechanismof antigen binding sites.