Noncovalent poly(1-vinylpyrrolidone)-based copolymer coating for the separation of basic proteins and lipoproteins by CE

✍ Scribed by Ai-Jun Wang; Joanna Witos; Lucia D'Ulivo; Kati Vainikka; Marja-Liisa Riekkola

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 532 KB
• Volume: 30
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.200900395

✦ Synopsis

Abstract

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 α‐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.