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KMID : 0352519940310010017
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Korea Univercity Medical Journal
1994 Volume.31 No. 1 p.17 ~ p.40
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Application of Monoclonal Anti- -Glutamyltransferase Antibodies in Enzyme
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Abstract
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γ-Glutamyltransferase (GGT : E. C. 2. 3. 2. 2.) is a glycoprotein enzyme which is
involved in glutathione metabolism and amino acid transport through the plasma membrane. it is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be
associated
with
hepatocarcinogenesis.
We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT. Liver GGT from AAF treated rats was
biochemically purified to a specific activity of 125.6 units per mg of protein. The overall purification was 551 folds and the final yield was 20.8%
Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and
protein A sepharose CS-4B column chromatography.
Using these mAbs we performed enzyme linked immunosorbent assay (ELISA),
radioimmunoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study
the distribution of GGT isozymes in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kidney GGT. These mAbs may be used to evaluate
the
distribution of GGT isozymes in different tissues.
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