title: In situ delipidation of low-density lipoproteins in capillary electrochromatography yields apolipoprotein B-100-coated surfaces for interaction studies.
abstract: An electrochromatographic method was developed for the in situ delipidation of intact low-density lipoprotein (LDL) particles immobilized on the inner wall of a 50-microm inner diameter silica capillary. In this method, the immobilized LDL particles were delipidated with nonionic surfactant Nonidet P-40 at pH 7.4 and 25 degrees C, resulting in an apolipoprotein B-100 (apoB-100)-coated capillary surface. The mobility of the electroosmotic flow marker dimethyl sulfoxide gave information about the surface charge, and the retention factors of beta-estradiol, testosterone, and progesterone were informative of the surface hydrophobicity. The calculated distribution coefficients of the steroids produced specific information about the affinity interactions of the steroids, with capillary surfaces coated either with intact LDL particles or with apoB-100. Delipidation with Nonidet P-40 resulted in a strong decrease in the hydrophobicity of the LDL coating. Atomic force microscopy images confirmed the loss of lipids from the LDL particles and the presence of apoB-100 protein coating. The in situ delipidation of LDL particles in capillaries represents a novel approach for the isolation of immobilized apoB-100 and for the determination of its pI value. The technique requires extremely low quantities of LDL particles, and it is simple and fast.
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, University of Helsinki, P.O. Box 55, FIN-00014 Helsinki, Finland.
authors.forename: Lucia
authors.initials: L
authors.lastname: D'Ulivo
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Meinander
authors.forename: Katariina
authors.initials: K
authors.lastname: Oörni
authors.forename: Petri T
authors.initials: PT
authors.lastname: Kovanen
authors.forename: Marja-Liisa
authors.initials: ML
authors.lastname: Riekkola
mesh_terms: D053299:Apolipoprotein B-100 / Q000737:chemistry*; D053801:Capillary Electrochromatography / Q000379:methods*; D006801:Humans; D007526:Isoelectric Point; D008077:Lipoproteins, LDL / Q000032:analysis / Q000097:blood*
chemical_list: D053299:Apolipoprotein B-100; D008077:Lipoproteins, LDL