Affinity electrophoresis
[1] The methods are based on changes in the electrophoretic pattern of molecules (mainly macromolecules) through biospecific interaction or complex formation.
Using a low voltage (~10 V/cm) to minimize the risk for heat damage, electricity is run across an agarose gel.
[citation needed] This technique utilizes a high voltage (≥ 20 V/cm) with a 0.5× Tris-borate buffer run across an agarose gel.
[10] Affinity capillary electrophoresis (ACE) refers to a number of techniques which rely on specific and nonspecific binding interactions to facilitate separation and detection through a formulary approach in accordance with the theory of electromigration.
[15] With ACE, scientists hope to develop strong binding drug candidates, understand and measure enzymatic activity, and characterize the charges on proteins.
[13] It is assumed for dynamic equilibrium affinity capillary electrophoresis that ligand-receptor binding occurs rapidly when the analyte and buffer are mixed.
Binding constants are generally derived from this technique based upon the peak migration shift of the receptor which is dependent upon the concentration of the analyte in the sample.
[18] CEC offers the highest separation efficacy of all three ACE techniques as non-matrixed sample components are washed away and the ligand then be released and analyzed.
[16] ACE is advantageous because it has a high separation efficiency, has a shorter analysis time, can be run at physiological pH, and involves low consumption of ligand/molecules.
[20] Affinity-trap polyacrylamide gel electrophoresis (PAGE) has become one of the most popular methods of protein separation.
This is not only due to its separation qualities, but also because it can be used in conjunction with a variety of other analytic methods, such as mass spectrometry, and western blotting.
