Noncovalent poly(1-vinylpyrrolidone)-based copolymer coating for the separation of basic proteins and lipoproteins by CE.
A new simple and fast noncovalent coating method based on poly(1-vinylpyrrolidone-co-2-dimethylaminoethyl methacrylate) copolymer was developed for CE. Merely 2 min flushing of the capillary with the poly(1-vinylpyrrolidone-co-2-dimethylaminoethyl methacrylate) copolymer was required. The copolymer is adsorbed onto the fused-silica surface by hydrogen bonding and electrostatic interactions. EOF was almost totally suppressed over a wide pH range. The coating conditions (flushing time, copolymer concentration, and the concentration and pH of background electrolyte solution) and the stability of the coating were optimized, and the coated capillary was successfully applied to the fast separation of four basic proteins: lysozyme, cytochrome c, ribonuclease A, and alpha-chymotrypsinogen A. Separation efficiencies were high, ranging from 386 000 to 738 000 plates/m at 40 mM pH 4.0 acetate buffer being comparable to values obtained on classical covalent PVP-coated capillary. The RSD of migration times of basic proteins for 200 times successive runs were all below 1.0% (n=200, 3 days). A successful capillary performance was demonstrated also to the separation of low- and high-density lipoproteins at acidic pH.
Wang
Ai-Jun
AJ
Laboratory of Analytical Chemistry, Department of Chemistry, University of Helsinki, Finland.
Witos
Joanna
J
D'Ulivo
Lucia
L
Vainikka
Kati
K
Riekkola
Marja-Liisa
ML
D000327:Adsorption; D000818:Animals; D002919:Chymotrypsinogen / Q000302:isolation & purification; D045304:Cytochromes c / Q000302:isolation & purification; D004355:Drug Stability; D053841:Electroosmosis; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008074:Lipoproteins / Q000302:isolation & purification*; D009113:Muramidase / Q000302:isolation & purification; D011109:Polymethacrylic Acids / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D011760:Pyrrolidinones / Q000737:chemistry*; D015203:Reproducibility of Results; D012259:Ribonuclease, Pancreatic / Q000302:isolation & purification
D008074:Lipoproteins; D011109:Polymethacrylic Acids; D011506:Proteins; D011760:Pyrrolidinones; C547200:poly(1-vinylpyrrolidone-co-2-dimethylaminoethyl methacrylate); D045304:Cytochromes c; D002919:Chymotrypsinogen; D012259:Ribonuclease, Pancreatic; D009113:Muramidase