A simplified procedure for the purification of human N-acetylglucosaminyltransferase-III from human hepatocellular carcinoma tissues

Human N-acetylglucosaminyltransferase-III (GnT-III) has not been hitherto p urified from either tissue or cell line. Although rat GnT-III has been puri fied from rat kidney tissue, the procedure involves laborious and complex s teps. We have developed a simplified procedure based on a QAE (diethyl[2-hy droxypropyl]aminoethyl)-Sepharose and an immunoaffinity chromatography tech nique. The immunoaffinity chromatography utilized a monoclonal antibody to human aglycosyl recombinant N-acetylglucosaminyltransferase-III. With the n ew procedure, human N-acetylglucosaminyltransferase-III was purified from h epatocellular carcinoma tissues with an increase in the specific enzyme act ivity of 378-fold.