Transformable capillary electrophoresis for oligosaccharide separations using phospholipid additives.
Phospholipids are used as an additive in capillary electrophoresis to enhance the separation of glycans derived from alpha1-acid glycoprotein, fetuin, and ribonuclease B. The properties of phospholipid preparations are dependent upon composition, hydration, and temperature. Separation performance is evaluated as a function of these variables. A preparation of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), with [DMPC]/[DHPC] = 2.5, in 10% lipid/aqueous buffer at 25 degrees C provides the best separation efficiency at an electric field strength of 400 V/cm. Resolution is enhanced with the additive. Concanavalin A, a lectin selective for high mannose and mannose branching glycans, and alpha1-2,3 mannosidase, an enzyme that cleaves 1-2 and 1-3 mannopyranosyl residues, are incorporated in the separation to provide additional selectivity and to expand the application of phospholipid additives for glycan separation.
Luo
Ruijuan
R
C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, USA.
Archer-Hartmann
Stephanie A
SA
Holland
Lisa A
LA
D000818:Animals; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry; D008361:Mannosidases / Q000378:metabolism; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry*; D015203:Reproducibility of Results; D013696:Temperature
D006023:Glycoproteins; D009844:Oligosaccharides; D010743:Phospholipids; D008361:Mannosidases