Urinary protein analysis.
The differentiation and analysis of urinary proteins has substantially contributed to our knowledge of physiological and pathophysiological processes during glomerular filtration and tubular catabolism of plasma constituents. By use of high-resolution biochemical separation techniques, several urinary polypeptides could be identified as plasma proteins, tissue antigens, tubular enzymes and protein breakdown products. With regard to clinical application, the separation results of conventional gel chromatography and agarose electrophoresis were surpassed by fast protein liquid chromatography and polyacrylamide electrophoresis in one- and two-dimensional systems. In contrast to early one-dimensional polyacrylamide gel electrophoresis (PAGE) methods using homogeneous gels on a macro scale, modern gradient slab gels achieve better resolution over the entire relative molecular mass range of urinary proteins. For clinical demands, the use of micro-scale gradient gels, either laboratory-made or pre-cast, together with an improved Coomassie Brillant Blue staining, offers several advantages, including rapidity, sensitivity and economy. Isoelectric focusing and two-dimensional PAGE, combined with sensitive silver staining and immunoblotting methods, have proved to be valuable tools for the identification and characterization of urinary proteins in defined renal and extra-renal diseases. The quantitative determination of urinary indicator proteins such as albumin and alpha 1- and beta 2-microglobulin can be regarded as a reasonable complement to the pattern diagnosis, especially in the long-term course of renal diseases.
Weber
M H
MH
Zentrum Innere Medizin der Universität Göttingen, F.R.G.
D000818:Animals; D002845:Chromatography; D004586:Electrophoresis; D006801:Humans; D011507:Proteinuria / Q000652:urine*