Phospholipid bilayer coatings for the separation of proteins in capillary electrophoresis.
The double-chained, zwitterionic phospholipid 1,2-dilauroyl-sn-phosphatidylcholine (DLPC, C12) was investigated for its use as a wall coating for the prevention of protein adsorption in capillary electrophoresis. DLPC forms a semipermanent coating at the capillary wall, which allows excess phospholipid to be removed from the capillary prior to electrophoretic separation. A DLPC-coated capillary allowed for the separation of both cationic and anionic proteins with efficiencies as high as 1.4 million plates/m. Migration time reproducibility was less than 1.3% RSD from run to run and less than 4.0% RSD from day to day. Protein recovery was as high as 93%. Cationic and anionic proteins could be separated over a pH range of 3-10, all yielding good efficiencies (N up to 1 million plates/m). The chain length of the phospholipid affected the performance of the wall coating. The C10 analogue of DLPC (DDPC) did not form a coating on the capillary wall while the C14 analogue of DLPC (DMPC) formed a stable coating that prevented protein adsorption to the same extent as its C12 counterpart.
Cunliffe
Jennifer M
JM
Department of Chemistry, University of Alberta, Edmonton, Canada.
Baryla
Nicole E
NE
Lucy
Charles A
CA
D004527:Egg Proteins / Q000302:isolation & purification; D019075:Electrophoresis, Capillary; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers; D010713:Phosphatidylcholines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D004527:Egg Proteins; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010743:Phospholipids; D011506:Proteins