Part I highlights the mechanisms of glomerular filtration and tubular reabsorption of plasma proteins, selected characteristics of urinary proteins based upon electrophoretic properties and recent advances in clinical laboratory analysis of proteinuria. Both structural characteristic of the glomerular capillary wall and molecular properties of plasma proteins are important determinants of glomerular filtration. Proteins filtered by the glomerulus subsequently appear in urine only after escaping the efficient mechanisms of tubular reabsorption. Albumin is one such protein and constitutes the major protein in normal urine although trace amounts of alpha, beta, and gamma globulins are also detectable. Several techniques of protein analysis have thus been developed to specifically measure albumin as well as other plasma proteins. Other methods have been adapted to measure total urinary protein content enabling the clinician to readily monitor renal function in health and disease. The second part of this review will consider conditions associated with proteinuria in both asymptomatic individuals and patients with renal disease. Asymptomatic proteinuria encompasses states of excess protein excretion during conditions of orthostasis, exercise, travel to high altitude of fever. Proteinuria during renal disease has received considerable interest as a means to monitor kidney function. It is therefore classified according to the type of damage incurred: (1) glomerular-type where large molecular weight proteins are excreted (2) tubular-type where small molecular weight proteins are excreted and (3) mixed-type characterized by both large and small molecular weight proteinuria.

Normal and abnormal aspects of proteinuria: Part I: Mechanisms, characteristics and analyses of urinary protein. Part II: Clinical considerations

We describe a simplified electroimmunoassay for quantification of human apolipoproteins A-I and B on prepared plates. A solution of agarose at 55 degrees C, containing hydroxyethylcellulose and antibodies, is poured onto a plastic film and allowed to gel. Wells are punched in the gels and the plates are dried for storage. Before use, they are rehydrated and buffered. We use succinylated Sudan Black to prestain lipoprotein fractions of plasma. After electrophoresing samples of plasma or standards for 3 h at 4 degrees C at 12.5 V/cm, we measure the peak heights and read the results from a standard curve prepared by using calibrated sera of known apolipoprotein B and A-I content as secondary standard. The within- and between-assay coefficients of variation were less than 4% in all cases. Results correlated well with those obtained by classic electroimmunodiffusion. Subjects with confirmed atherosclerotic lesions had significantly (p less than 10(-9)) lower ratios of apolipoprotein A-I to apolipoprotein B, compared with ratios in controls.

Simultaneous measurement of plasma apolipoproteins A-I and B by electroimmunoassay.

The application of isoelectric focusing (IEF), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional electrophoresis (2-DE) and capillary electrophoresis (CE) for high resolution electrophoretic analysis of human urinary proteins is reviewed. In each case, the information is tabulated chronologically with details of sample preparation, electrophoretic system, detection method and clinical application. The text includes an historical perspective of the use of each method for urinalysis and a detailed review of the application of the methods to the investigation of renal disease, renal transplantation, Bence Jones proteinuria (BJP), diabetes mellitus, cadmium toxicity, nephrolithiasis and cancers of the urogenital tract.

Clinical analysis of human urinary proteins using high resolution electrophoretic methods.

Urinary protein analysis.

The electrophoresis of Bence Jones proteinuria (BJP) by urinary protein electrophoresis (UPE), immunoelectrophoresis (IE), immunofixation electrophoresis (IFE), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF), two-dimensional electrophoresis (2-DE) and capillary electrophoresis (CE) is described. UPE, IE and IFE are briefly discussed as clinical laboratory methods for the detection and typing of free light chain (LC) whilst the high resolution electrophoretic methods (SDS-PAGE, IEF, 2-DE and CE) are considered in greater detail as research tools for molecular characterisation of free LC and its association with nephrotoxicity. Refinements of sample processing designed to improve the standardisation of analysis of BJP by high resolution electrophoretic methods are reported.

Electrophoretic analysis of bence jones proteinuria

We describe a procedure for the convenient separation of proteins by sodium dodecyl sulfate/polyacrylamide gel electrophoresis of urine from cases of renal disease. A precipitation method that requires no special apparatus was used to concentrate the urinary proteins; for electrophoretic separation we used a commercially supplied polyacrylamide/cellulose gel slab. This method seems to be valuable for investigation of proteinuria; we recommend it for routine use.

Rapid sodium dodecyl sulfat/polyacrylamide gel electrophoresis of urinary proteins.

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Rapid sodium dodecyl sulfat/polyacrylamide gel electrophoresis of urinary proteins.