title: Rapid purification of beta-glucuronidase from human liver by immunoaffinity chromatography employing specific murine monoclonal IgG binding to tresyl-activated agarose.
abstract: We have developed a simple, rapid method for purification of beta-glucuronidase from human liver in order to facilitate the study of its biochemical structure and pathophysiologic roles in both cholelithiasis and carcinogenesis. The procedure includes the following steps: (1) liver homogenization, (2) 25-45% saturated ammonium sulfate fractionation, (3) heat denaturation, and (4) immunoaffinity chromatography employing murine anti-human beta-glucuronidase monoclonal IgG binding to tresyl-activated agarose. beta-Glucuronidase constitutes 1.3 mg per 100 g of wet liver tissue. The enzyme can be purified with a 10% overall yield and overall purification of 5000-fold in a 2-day cycle on a fairly large scale by the method described. Polyacrylamide gel electrophoresis indicated minor contaminants in the final product which could be further purified by protein blotting.
authors.affiliation: Department of Pathology, University of Alabama, Birmingham.
authors.forename: K J
authors.initials: KJ
authors.lastname: Ho
mesh_terms: D000645:Ammonium Sulfate; D000911:Antibodies, Monoclonal; D005591:Chemical Fractionation; D002846:Chromatography, Affinity / Q000379:methods*; D005966:Glucuronidase / Q000276:immunology / Q000302:isolation & purification*; D006358:Hot Temperature; D006801:Humans; D007074:Immunoglobulin G; D008099:Liver / Q000201:enzymology*; D011489:Protein Denaturation; D012685:Sepharose
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D012685:Sepharose; D005966:Glucuronidase; D000645:Ammonium Sulfate