Highly efficient protein separations in capillary electrophoresis using a supported bilayer/diblock copolymer coating.
A surfactant/polymer wall coating consisting of the doubly chained cationic surfactant dimethyldioctadecylammonium bromide (DODAB) and polyoxyethylene (POE) 40 stearate is investigated. The coating is formed by simply rinsing a capillary with a solution containing DODAB and POE 40 stearate. The resultant coating is semi-permanent--demonstrating stable electroosmotic flow (EOF) even after a 60 min high pressure rinse with buffer. The EOF (-0.45+/-(0.23) x 10(-4) cm(2) V(-1) s(-1) at pH 7.4) is suppressed by more than a factor of ten compared to that observed for DODAB alone. Model protein mixtures were separated over a pH range of 3-10 with efficiencies of up to greater than 1 million plates/m for the basic proteins cytochrome c, lysozyme, ribonuclease A and alpha-lactalbumin, and the acidic proteins insulin chain A, trypsin inhibitor, and alpha-chymotrypsinogen A. Migration time reproducibility was 0.5-4.0% from run to run and 0.6-4.3% from day to day. Protein recoveries with this coating ranged from 84% to 97%.
MacDonald
Amy M
AM
Department of Chemistry, University of Alberta, Gunning/Lemieux Chemistry Centre, Edmonton, Alta., Canada T6G 2G2.
Lucy
Charles A
CA
D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D015203:Reproducibility of Results
D011108:Polymers; D011506:Proteins; D000644:Quaternary Ammonium Compounds; C015831:dimethyldioctadecylammonium