Evaluation of commonly used electrophoretic methods for the analysis of proteins and peptides and their application to biotechnology

Abstract

This account gives an overview and evaluation of the current methods of most commonly used electrophoretic techniques in the analysis of various polypeptides. A general review of some background information and pitfalls associated with the application of these methods in the characterization of the size and charge properties of proteins or peptides is presented. It is intended to provide some guidelines to those researchers first exposed to this important and powerful methodology in modern life sciences. Electrophoretic characterizations of several major classes of proteins with vastly different molecular sizes and/or charges are taken as exemplars to highlight the versatility of electrophoretic protocols and their great potential for facilitating protein purification and structural analysis of some complex and scarce biological samples when coupled to other modern analytical methods. Major electrophoretic techniques reviewed herein include: 1. Discontinuous polyacrylamide gel electrophoresis (DISC-PAGE) 2. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 3. Pore gradient electrophoresis (PGE) 4. Isoelectric focusing (IEF) and 5. Two-dimensional gel electrophoresis (2D PAGE).