title: Immunoprecipitation of apolipoprotein B-containing lipoproteins for isolation of HDL particles.
abstract: BACKGROUND
  Immunoprecipitation (IP) of non-HDL particles with antisera provides the simplest and most specific method available for the separation of HDL. We compared the LipoSep™ IP reagent with the dextran sulfate/MgCl2 precipitation method (DS).
  
  
  METHODS
  The IP reagent (200 μl) was added to an equal volume of serum, vortexed, incubated for 10 min at room temperature, and microcentrifuged at 12,000 rpm for 10 min.
  
  
  RESULTS
  Equal volumes of a sample and the IP reagent precipitated apoB to 3.0 g/l without the coprecipitation of HDL. HDL-C measured in the supernatant after IP (Y) gave excellent agreement to DS precipitation (X) with a slope of 1.01, an intercept of 0.070 mmol/l (2.7 mg/dl), and a correlation of 0.99 (n=118; apoB 0.16-2.11 g/l). However, DS failed in most samples with moderate to elevated triglycerides. At triglyceride concentrations from 2.86 to 23.63 mmol/l (253-2091 mg/dl) the initial success rate was 65.4% for IP, while DS successfully precipitated only 5.8%. Success rate on repeat with additional reagent and/or sample dilution gave a success rate of 86.5% for IP and 40.4% for DS.
  
  
  CONCLUSIONS
  The IP reagent and protocol is a simple, effective and highly specific tool for isolating HDL particles in human serum and is effective with high triglyceride samples.
authors.affiliation: Sun Diagnostics, LLC, 60 Pineland Drive, Brunswick Hall, Suite 322, New Gloucester, ME 04260, USA. Electronic address: jcontois@sundiagnostics.us.
authors.forename: John H
authors.initials: JH
authors.lastname: Contois
authors.affiliation: Sun Diagnostics, LLC, 60 Pineland Drive, Brunswick Hall, Suite 322, New Gloucester, ME 04260, USA.
authors.forename: Andre L
authors.initials: AL
authors.lastname: Albert
authors.affiliation: Sun Diagnostics, LLC, 60 Pineland Drive, Brunswick Hall, Suite 322, New Gloucester, ME 04260, USA.
authors.forename: Rae-Anne
authors.initials: RA
authors.lastname: Nguyen
mesh_terms: D001055:Apolipoproteins B / Q000737:chemistry*; D002498:Centrifugation; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D008075:Lipoproteins, HDL / Q000737:chemistry* / Q000302:isolation & purification*; D013997:Time Factors
chemical_list: D001055:Apolipoproteins B; D008075:Lipoproteins, HDL
keywords: Cholesterol; High density lipoprotein; Immunoprecipitation; Lipoproteins