title: The effects of apolipoprotein B depletion on HDL subspecies composition and function.
abstract: HDL cholesterol (HDL-C) efflux function may be a more robust biomarker of coronary artery disease risk than HDL-C. To study HDL function, apoB-containing lipoproteins are precipitated from serum. Whether apoB precipitation affects HDL subspecies composition and function has not been thoroughly investigated. We studied the effects of four common apoB precipitation methods [polyethylene glycol (PEG), dextran sulfate/magnesium chloride (MgCl2), heparin sodium/manganese chloride (MnCl2), and LipoSep immunoprecipitation (IP)] on HDL subspecies composition, apolipoproteins, and function (cholesterol efflux and reduction of LDL oxidation). PEG dramatically shifted the size distribution of HDL and apolipoproteins (assessed by two independent methods), while leaving substantial amounts of reagent in the sample. PEG also changed the distribution of cholesterol efflux and LDL oxidation across size fractions, but not overall efflux across the HDL range. Dextran sulfate/MgCl2, heparin sodium/MnCl2, and LipoSep IP did not change the size distribution of HDL subspecies, but altered the quantity of a subset of apolipoproteins. Thus, each of the apoB precipitation methods affected HDL composition and/or size distribution. We conclude that careful evaluation is needed when selecting apoB depletion methods for existing and future bioassays of HDL function.
authors.affiliation: Center for Lipid and Arteriosclerosis Science, Department of Pathology and Laboratory Medicine, University of Cincinnati, Cincinnati, OH 45237.
authors.forename: W Sean
authors.initials: WS
authors.lastname: Davidson
authors.affiliation: Department of Pediatrics, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH 45229.
authors.forename: Anna
authors.initials: A
authors.lastname: Heink
authors.affiliation: Department of Pediatrics, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH 45229.
authors.forename: Hannah
authors.initials: H
authors.lastname: Sexmith
authors.affiliation: Center for Lipid and Arteriosclerosis Science, Department of Pathology and Laboratory Medicine, University of Cincinnati, Cincinnati, OH 45237.
authors.forename: John T
authors.initials: JT
authors.lastname: Melchior
authors.affiliation: National Heart, Lung, and Blood Institute, Lipoprotein Metabolism Section,, Bethesda, MD 20892.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Gordon
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Kuklenyik
authors.affiliation: Center for Lipid and Arteriosclerosis Science, Department of Pathology and Laboratory Medicine, University of Cincinnati, Cincinnati, OH 45237.
authors.forename: Laura
authors.initials: L
authors.lastname: Woollett
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341.
authors.forename: John R
authors.initials: JR
authors.lastname: Barr
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341.
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Jones
authors.affiliation: Battelle Memorial Institute, Analytical Services, Atlanta, GA 30329.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Toth
authors.affiliation: Department of Pediatrics, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH 45229 amy.shah@cchmc.org.
authors.forename: Amy S
authors.initials: AS
authors.lastname: Shah
mesh_terms: D000328:Adult; D016632:Apolipoprotein A-I / Q000378:metabolism; D001055:Apolipoproteins B / Q000172:deficiency* / Q000302:isolation & purification* / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D011232:Chemical Precipitation / Q000187:drug effects; D002712:Chlorides / Q000494:pharmacology; D008076:Cholesterol, HDL / Q000737:chemistry / Q000378:metabolism; D016264:Dextran Sulfate / Q000494:pharmacology; D005260:Female; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000378:metabolism*; D017895:Manganese Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D010316:Particle Size; D011092:Polyethylene Glycols / Q000494:pharmacology
chemical_list: D016632:Apolipoprotein A-I; D001055:Apolipoproteins B; D002712:Chlorides; D008076:Cholesterol, HDL; D008077:Lipoproteins, LDL; D017895:Manganese Compounds; D011092:Polyethylene Glycols; D006493:Heparin; D016264:Dextran Sulfate; C025340:manganese chloride
keywords: cholesterol efflux; dextran sulfate; heparin sodium; high density lipoprotein; lipoproteins; low density lipoprotein; polyethylene glycol; proteomics