Urinary bladder glycoproteins of the rabbit: extraction, biochemical and immunological studies

• Published in: Immunological investigations Vol. 14; no. 1; p. 41
• Main Authors: Callahan, H J, Byrne, D, Fritz, R, Mulholland, S G
• Format: Journal Article
• Language: English
• Published: England 01.01.1985
• Subjects: Animals; Enzyme-Linked Immunosorbent Assay; Female; Glycoproteins - immunology; Glycoproteins - isolation & purification; Glycoproteins - physiology; Mice; Mucins - analysis; Mucus - analysis; Rabbits; Radioimmunoassay; Urinary Bladder - analysis
• ISSN: 0882-0139; 1532-4311
• DOI: 10.3109/08820138509052445

The mucin layer of the bladder covering the transitional epithelium is thought to be an anti-adherence substance for bacteria. We have previously demonstrated that removal of this layer results in increased bacterial colonization. In this present communication we report our attempts to isolate and characterize the components of the mucin layer. Bladders were removed from female NZW rabbits and the mucosal layer was extracted with 0.1 M NaCl. Fractional centrifugation and column chromatography on AcA 34 and AcA 22 resulted in the separation of a partially purified glycoprotein, containing 30% carbohydrate, migrating as a single peak (4.5S) in the analytical ultracentrifuge. It was immunogenic in mice and the antisera were used to develop both a radioimmunoassay and an enzyme linked immunosorbent assay (ELISA). The antigenic activity of the glycoprotein was destroyed by protease or extremes of pH, but not by several glycosidases. In addition the murine antisera could not be inhibited by a panel of naturally occurring glycoproteins or glycosaminoglycans.