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2 dimensional electrophoresis method for additional characterization of para proteins in serum clinical application of 2 dimensional electrophoresis 1



2 dimensional electrophoresis method for additional characterization of para proteins in serum clinical application of 2 dimensional electrophoresis 1



Journal of Clinical Chemistry & Clinical Biochemistry 22(1): 53-60



High resolution 2 dimensional electrophoresis, one of the most potent methods for the analytical separation of proteins, is still used only in specialized laboratories. A 2 dimensional electrophoresis method is described which permits characterization of abnormal [human] serum proteins (paraproteins) and is suitable for application in a clinical laboratory. Both steps of the electrophoresis are performed by the flat bed technique. The critical transfer from the isoelectric focusing step to the gel of the second dimension is facilitated by use of specially prepared foil stencils. The practicality of the procedure was considerably increased by using commercially available materials such as gels for isoelectric focusing and the silver stain kit. The analysis can be calibrated by measurement of the pH gradient and by using MW protein standards. Moreover, 2 different samples can be analyzed simultaneously in the same gels of the first and the second dimension. This permits a direct comparison of the sample of interest with the sample of reference. Serum samples showing an abnormal band (M gradient) in routine methods such as agarose gel electrophoresis, were analyzed by the described method. Some M gradients consisted of single abnormal proteins. Another sample showed a M gradient composed of a number of abnormal protein spots in this method. Questions concerning pathobiological and clinical evaluation of the findings justify further studies using this method.

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Accession: 004535205

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