Abstract
Affinity electrophoresis is an important technique that is widely used to separate and analyze biomolecules in the fields of biology and medicine. Both quantitative and qualitative information can be gained through affinity electrophoresis. Affinity electrophoresis can be applied through a variety of strategies, such as mobility shift electrophoresis, charge shift electrophoresis or capillary affinity electrophoresis. These strategies are based on changes in the electrophoretic patterns of biological macromolecules that result from interactions or complex-formation processes that induce changes in the size or total charge of the molecules. Nucleic acid fragments can be characterized through their affinity to other molecules, for example transcriptional factor proteins. Hydrophobic membrane proteins can be identified by means of a shift in the mobility induced by a charged detergent. The various strategies have also been used in the estimation of association/disassociation constants. Some of these strategies have similarities to affinity chromatography, in that they use a probe or ligand immobilized on a supported matrix for electrophoresis. Such methods have recently contributed to profiling of major posttranslational modifications of proteins, such as glycosylation or phosphorylation. Here, we describe advances in analytical techniques involving affinity electrophoresis that have appeared during the last five years.
Highlights
Affinity electrophoresis is a separation technique that is based on the phenomenon whereby the mobilities of charged molecules in a direct-current electric field vary according to whether or not they interact with other molecules for which they have affinities
Affinity electrophoresis techniques based on the retardation of the migration of molecules that bind strongly to affinity probes immobilized on a supported matrix have been developed, and these have contributed to analyses of major posttranslational modifications of proteins [11,12,13,14]
Polyacrylamide gel electrophoresis (PAGE) has become the most-widely adopted method for the separation of proteins, because it has good separation abilities and because it can be used in combination with various other analytical methods, such as Western blotting, amino acid sequence analysis or mass spectrometry, once the proteins have been transferred to the PVDF membrane
Summary
Affinity electrophoresis is a separation technique that is based on the phenomenon whereby the mobilities of charged molecules in a direct-current electric field vary according to whether or not they interact with other molecules for which they have affinities. A wide variety of affinity electrophoresis techniques have been developed that provide good qualitative results or have good separation capabilities. One such pioneering technique is that of affinophoresis [10], which utilizes an affinophore, a charged hydrophilic polymer that binds to molecules with a suitable affinity. Affinity electrophoresis techniques based on the retardation of the migration of molecules that bind strongly to affinity probes immobilized on a supported matrix have been developed, and these have contributed to analyses of major posttranslational modifications of proteins [11,12,13,14]. It introduces a technique for the analysis of phosphorylated proteins that is based on the phosphate-affinity probe, Phos-tag [11,26,27,28,29,30,31,32,33], which we developed, and it describes the contributions made to phosphoproteomics by techniques based on Phos-tag
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
