APPLICATIONS OF 6 MONOCLONAL-ANTIBODIES IN IN-VITRO AND IN-VIVO ASSAYS TO EVALUATE THE TISSUE-SPECIFICITY OF GAMMA-GLUTAMYL-TRANSFERASE

초록

We have purified gamma-glutamyltransferase (GGT) from rat liver which had been treated with 2-acetamidofluorene. Hybridomas were prepared against the purified GGT and six clones producing monoclonal antibodies (MAbs) were screened. All of these six MAbs were extensively subcloned and produced in large quantities by injecting these clones into the peritoneal cavity of BALB/c mice. From the ascitic fluid MAbs were then purified by ammonium sulfate precipitation followed by protein A-Sepharose affinity chromatography. In order to investigate specificity of these MAbs, we performed both in vivo assay of immunoscintigraphy and in vitro assays of enzyme-linked immunosorbent assay, radioimmunoassay, immunohistochemistry, and autoradiography. The result of these experiments indicated that GGT MAb-1, 2, 3 and 4 exhibit immunoreactivity against the rat Liver GGT induced by 2-acetamidofluorene, while GGT MAb-5 reacts with the normal rat liver GGT, and GGT MAb-6 with the normal kidney GGT. Therefore, these MAbs may be useful for detecting the distribution of GGT isozymes in different tissues and organs.

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