Simultaneous separation of cationic and anionic proteins using zwitterionic surfactants in capillary electrophoresis.
The zwitterionic surfactant Rewoteric AM CAS U forms a dynamic wall coating that prevents the adsorption of cationic proteins as well as suppresses the electroosmotic flow (EOF). Addition of polarizable anions to buffers containing this zwitterionic surfactant increases the once suppressed EOF to values nearing +3 x 10(-4) cm2/(V s). The retention of the EOF allows for the separation of analytes of widely different mobilities and is demonstrated by the simultaneous separation of cationic and anionic proteins. Using a buffer containing optimal amounts of the polarizable anion perchlorate and surfactant CAS U, the proteins lysozyme, ribonuclease A, alpha-chymotrypsinogen A, and myoglobin are separated in less than 15 min. Efficiencies as high as 1.5 million plates/m and recoveries greater than 91% are observed for proteins injected in distilled water. Migration time reproducibility is approximately 1% RSD within 1 day and approximately 3% RSD from day to day. The anionic and cationic proteins can be separated over a pH range of 5.5-9, all yielding good efficiencies.
Baryla
N E
NE
Department of Chemistry, University of Alberta, Edmonton, Canada.
Lucy
C A
CA
D000818:Animals; D000838:Anions; D001622:Betaine / Q000031:analogs & derivatives*; D002412:Cations; D002417:Cattle; D002919:Chymotrypsinogen / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods; D009113:Muramidase / Q000302:isolation & purification; D009211:Myoglobin / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic / Q000302:isolation & purification
D000838:Anions; D002412:Cations; D009211:Myoglobin; D011506:Proteins; C400714:amidopropylhydroxydimethylsulfobetaine; D001622:Betaine; D002919:Chymotrypsinogen; D012259:Ribonuclease, Pancreatic; D009113:Muramidase