Oligomerized phospholipid bilayers as semipermanent coatings in capillary electrophoresis

Abstract

Double-chained surfactants form semipermanent coatings that prevent protein adsorption in capillary electrophoresis (CE). To make such coatings more permanent, vesicles of the unsaturated phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine were prepared and subjected to free-radical-initiated polymerization, both inside the capillary and in free solution. The latter generated oligomers of 2-5 units based on ESI-TOF MS, and formed the more stable coating in CE. Rinsing the capillary with a solution of the ex situ oligomerized DOPC suppressed EOF (0.8 x 10(-)(8) m(2)/V.s) for more than 20 h, whereas in situ oligomerized electroosmotic flow (EOF) suppressed the EOF for only 10 h. Mixtures of anionic and cationic proteins were separated under neutral pH and low ionic strength buffer with efficiencies of 480,000-930,000 plates/m and recoveries of 75-99%.