Isolation and characterization of a human liver and kidney-specific protein: the hepato-renal (H-R) antigen.
This paper reports the isolation and characterization of a soluble antigen shared by the liver and kidney of human and some other animal species. Homogenates of human liver in saline were centrifugated at 27,000 g and the supernatants were fractionated by preparative polyacrylamide gel electrophoresis. The gels were divided in sections and each was injected into rabbits; after absorption with polymerized normal human serum, the antiserum obtained by injecting one of the sections reacted only with saline extracts of human liver and kidney when tested against a variety of human tissue extracts. The absorbed antiserum, polymerized and insolubilized with glutaraldehyde, was used to purify the antigen by affinity chromatography. The purified antigen proved to be a glycoprotein containing 19 percent carbohydrate, had a molecular weight of 5.8-6.0 x 10(4) Daltons and a pI of 7.2-7.4. The antigen, relatively thermostable, was precipitated by 35-55 percent ammonium sulphate; its antigenic activity was not affected by extraction with 0.6 N perchloric acid or by incubation with ribonuclease, deoxyribonuclease or neuraminidase but was destroyed by incubation with ttypsin or chymotrypsin. Immunoperoxidase studies showed that the antigen appeared concentrated in the neclei of liver and kidney glomerular epithelial and tubular epithelial cells in humans and rats. The antigen could not be detected in human hepatomas or hypernephromas or in the rat Morris hepatoma 5123.
D000230:Adenocarcinoma / Q000276:immunology; D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000302:isolation & purification*; D006528:Carcinoma, Hepatocellular / Q000276:immunology; D002467:Cell Nucleus / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D007668:Kidney / Q000276:immunology*; D007680:Kidney Neoplasms / Q000276:immunology; D008099:Liver / Q000276:immunology*; D008113:Liver Neoplasms / Q000276:immunology; D008970:Molecular Weight; D051381:Rats
D000941:Antigens; D000939:Epitopes; D006023:Glycoproteins