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Clin Chim Acta. 2014 Sep 25;436:348-50. doi: 10.1016/j.cca.2014.06.017. Epub 2014 Jun 30.

Immunoprecipitation of apolipoprotein B-containing lipoproteins for isolation of HDL particles.

Author information

  • 1Sun Diagnostics, LLC, 60 Pineland Drive, Brunswick Hall, Suite 322, New Gloucester, ME 04260, USA. Electronic address: jcontois@sundiagnostics.us.
  • 2Sun Diagnostics, LLC, 60 Pineland Drive, Brunswick Hall, Suite 322, New Gloucester, ME 04260, USA.

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.

CONCLUSION:

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.

Copyright © 2014 Elsevier B.V. All rights reserved.

KEYWORDS:

Cholesterol; High density lipoprotein; Immunoprecipitation; Lipoproteins

PMID:
24992525
[PubMed - indexed for MEDLINE]
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