Hydrophobic Interaction Chromatography of Proteins on Neutral Adsorbents
The separation mechanisms of most of the high-resolution biochemical fractionation techniques are based mainly on one particular separation parameter: in nonsieving electrophoresis and ion exchange chromatography, the surface charge of the solute of interest; in ultracentrifugation, in chromatographic molecular sieving, and in pore-gradient electrophoresis, the molecular size; in isopycnic gradient centrifugation, the density; in hydroxylapatite chromatography, the number of available carboxylic (and phosphate) groups in the solute molecules (Bernardi and Kawasaki, 1968). Since the above methods utilize mainly one separation parameter, they often give reproducible results, and the separation patterns can in many cases be correlated to particular molecular properties of the fractionated substances. For this reason, these separation methods are used routinely in most biochemical research laboratories. However, two or more separation parameteres can with advantage be utilized simultaneously, provided that the contribution of each parameter can be kept constant from experiment to experiment, which is a prerequisite for reproducible results. In practice, it is often very difficult to fulfill this requirement. Polyacrylamide gel electrophoresis is an example of the few methods which utilize two separation parameters and still provide both high reproducibility and high resolution.
KeywordsGlycidyl Ether Hydrophobic Interaction Chromatography Cyanogen Bromide Separation Parameter Separation Pattern
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