Diagnostic use of an analysis of urinary proteins by a practicable sodium dodecyl sulfate‐electrophoresis method and rapid two‐dimensional electrophoresis

Abstract

Two methods suitable for routine clinical analyses of urinary proteins are presented and compared. The first is a horizontal sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique, suitable for simultaneous analysis of 20 native urinary samples. This method uses polyacrylamide gradient gels, prepared with a laboratory-built gel casting device. The second method is a rapid two-dimensional electrophoresis procedure, combining cellulose acetate electrophoresis and sodium dodecyl sulfate-electrophoresis. The first step uses a routine system (Chemetron), the second separation step followed by staining with Coomassie Brilliant Blue R is performed on the PhastSystem. The resulting two-dimensional patterns reveal urinary proteins distributed according to the 5-zone pattern of native proteins (albumin, alpha-1, alpha-2, beta, gamma-globulin) as well as to the logarithm of their molecular weights. Examples of (routine) diagnoses with a special interest in the monitoring of kidney transplant patients are shown.

Microalbuminuria and tubular proteinuria as risk predictors of cardiovascular morbidity and mortality in essential hypertension: final results of a prospective long-term study (MARPLE Study)*

“…The following parameters were assessed: creatinine using the method of Jaffé , albumin, alpha 1-microglobulin and total protein using the nephelometric method; urine electrophoresis using sodiumdodecyl sulphate (SDS) was also performed [19][20][21].…”

Peptide mapping of proteins in human body fluids using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

“…The plasma proteins have many important functions in the circulation, for example, stabilizing the salt-water balance and transportation of low-Mw compounds (for instance, hydrophobic substances; e.g., fatty acids; or substances that are toxic in free form; e.g., bilirubin). There are also a number of smaller proteins bound to plasma proteins to avoid excretion by glomerular filtration in the kidney (e.g., retinol-binding protein bound to transthyretin (Lapin, Gabl, & Kopsa, 1989;Marshall & Williams, 1998), and globulin dimers bound to haptoglobin (Marshall & Williams, 1987)). Furthermore, many of these proteins act as protectors of various systems, such as extracellular protease inhibitors, immunoglobulins, complement factors, and coagulation factors.…”

The simplified technique of high resolution two‐dimensional polyacrylamide gel electrophoresis: Biomedical applications in health and disease

“…[66] investigated first-dimensional IEF in immobilized pH gradients for large volume (up to 1 mL) analysis. Alternative 2-D PAGE methods have also been used, including nondenaturing conditions [22,67] and cellulose acetate electrophoresis followed by SDS-PAGE [68]. For an excellent review of urinary proteins and their electrophoretic analysis see Weber t691.…”