The separation of basic proteins by capillary electrophoresis with carboxymethyl chitosan coated cap

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  A hydrophilic basic polysaccharide, carboxymethyl chitosan (CMC) as capillary coating, which has been proved to be simple, stable and low cost, was utilized to separate some typical basic proteins.The separation of these proteins on uncoated capillary always displays a severe tailing due to the electrostatic attraction between the cationic proteins and anionic silica surface.In our study, several experimental parameters, such as the concentration and pH of the running buffer were optimized for the separation of basic proteins, and also compared with that of uncoated capillary.The results demonstrated that three basic proteins including cytochrome C, lysozyme and ribonuclease A, can be separated by CMC-coated capillary within 10 min (Fig.l) by using a buffer of 100mM HAc-NaAc (pH=4.0), 25℃, and applied voltage +30kV.The RSD of the migration time and area were lower than 1.0% and 4.0%, respectively (n=5), indicating that satisfactory reproducibility can be obtained on CMC-coated capillary for analysis of basic proteins.
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