title: Multidimensional profiling of plasma lipoproteins by size exclusion chromatography followed by reverse-phase protein arrays.
abstract: The composition of lipoproteins and the association of proteins with various particles are of much interest in the context of cardiovascular disease. Here, we describe a technique for the multidimensional analysis of lipoproteins and their associated apolipoproteins. Plasma is separated by size exclusion chromatography (SEC), and fractions are analyzed by reverse-phase arrays. SEC fractions are spotted on nitrocellulose slides and incubated with different antibodies against individual apolipoproteins or antibodies against various apolipoproteins. In this way, tens of analytes can be measured simultaneously in 100 μl of plasma from a single SEC separation. This methodology is particularly suited to simultaneous analysis of multiple proteins that may change their distribution to lipoproteins or alter their conformation, depending on factors that influence circulating lipoprotein size or composition. We observed changes in the distribution of exchangeable apolipoproteins following addition of recombinant apolipoproteins or interaction with exogenous compounds. While the cholesteryl ester transfer protein (CETP)-dependent formation of pre-β-HDL was inhibited by the CETP inhibitors torcetrapib and anacetrapib, it was not reduced by the CETP modulator dalcetrapib. This finding was elucidated using this technique.
authors.affiliation: Discovery Technologies, F. Hoffmann-La Roche, Ltd, Basel, Switzerland and.
authors.forename: Gregor
authors.initials: G
authors.lastname: Dernick
authors.affiliation: Discovery Technologies, F. Hoffmann-La Roche, Ltd, Basel, Switzerland and.
authors.forename: Stefan
authors.initials: S
authors.lastname: Obermüller
authors.affiliation: Discovery Technologies, F. Hoffmann-La Roche, Ltd, Basel, Switzerland and.
authors.forename: Cyrill
authors.initials: C
authors.lastname: Mangold
authors.affiliation: Discovery Technologies, F. Hoffmann-La Roche, Ltd, Basel, Switzerland and. Electronic address: gregor.dernick@roche.com.
authors.forename: Christine
authors.initials: C
authors.lastname: Magg
authors.affiliation: Discovery Technologies, F. Hoffmann-La Roche, Ltd, Basel, Switzerland and.
authors.forename: Hugues
authors.initials: H
authors.lastname: Matile
authors.affiliation: Discovery Technologies, F. Hoffmann-La Roche, Ltd, Basel, Switzerland and.
authors.forename: Oliver
authors.initials: O
authors.lastname: Gutmann
authors.affiliation: Metabolic and Vascular Diseases, F. Hoffmann-La Roche, Ltd, Basel, Switzerland.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: von der Mark
authors.affiliation: Metabolic and Vascular Diseases, F. Hoffmann-La Roche, Ltd, Basel, Switzerland.
authors.forename: Corinne
authors.initials: C
authors.lastname: Handschin
authors.affiliation: Metabolic and Vascular Diseases, F. Hoffmann-La Roche, Ltd, Basel, Switzerland.
authors.forename: Cyrille
authors.initials: C
authors.lastname: Maugeais
authors.affiliation: Metabolic and Vascular Diseases, F. Hoffmann-La Roche, Ltd, Basel, Switzerland.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Niesor
mesh_terms: D000918:Antibody Specificity; D016477:Artifacts; D001774:Blood Chemical Analysis / Q000379:methods*; D053480:Cholesterol Ester Transfer Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D002850:Chromatography, Gel / Q000379:methods*; D053442:High-Density Lipoproteins, Pre-beta / Q000097:blood / Q000378:metabolism; D006801:Humans; D008074:Lipoproteins / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011804:Quinolines / Q000494:pharmacology; D015203:Reproducibility of Results
chemical_list: C506642:CETP protein, human; D053480:Cholesterol Ester Transfer Proteins; D053442:High-Density Lipoproteins, Pre-beta; D008074:Lipoproteins; D011804:Quinolines; C483909:torcetrapib