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Electrophoretic, chromatographic and immunological studies of human urinary proteins
Author(s) -
Büeler Martin R.,
Wiederkehr Felix,
Vonderschmitt Dieter J.
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150160122
Subject(s) - urine , chemistry , blood proteins , prostatic acid phosphatase , urinary system , secretory component , chromatography , tamm–horsfall protein , affinity chromatography , fractionation , biochemistry , protein subunit , antibody , biology , enzyme , acid phosphatase , endocrinology , immunology , gene
Urinary proteins from both sexes were analyzed by high resolution two‐dimensional gel electrophoresis (2‐DE). For well reproducible 2‐DE patterns, the samples were concentrated and desalted in one step by vacuum dialysis. A reference map for urine proteins was established by the analysis of urine from 10 healthy persons. Proteins in urine that share immunogenicity with serum proteins were identified by use of antibody to whole human‐serum protein in an affinity‐column fractionation of urine and differential analysis of the adsorbed (serum component) and unadsorbed (non‐serum component) fractions. For identification of individual proteins, coelectrophoresis, immunoblotting and affinity chromatography with corresponding antibodies were used. Proteins identified in the map, besides known serum proteins, included: the subunit of Tamm‐Horsefall protein, the secretory component of IgA, constant breakdown products of αl‐antitrypsin and retinol‐binding protein, the five isoforms of the β chain of human chorionic gonadotropin and the subunit of prostatic acid phosphatase. In addition, we could demonstrate three proteins which are markedly pronounced in female urine, especially pregnant women. To get more information about the native properties of various urinary proteins, they were separated into four main peaks according to their sizes using fast protein liquid chromatography equipment. Possible interpolypeptide disulfide bonds were studied using a nonreducing 2‐DE system. 2‐DE in combination with other methods seems to be a valuable tool for the characterization of urinary proteins in defined renal or extra‐renal diseases. An example is given by analyzing the immune complexes from seven patients with a urinary tract infection.