A rapid, economical, and reproducible method for human serum delipidation and albumin and IgG removal for proteomic analysis

Qin Fu, Diane E. Bovenkamp, Jennifer E. Van Eyk

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Serum is a readily available source for diagnostic assays, but the identification of disease-specific serum, biomarkers has been impeded by the dominance of human serum albumin. (HSA) and immunoglobulin G (IgG) in the serum, proteome. Therefore, in order to observe lower-abundance serum proteins, removal or depletion of at least these two proteins is required. However, the depletion method needs to be inexpensive and reproducible. We describe such a protocol that combines delipidation by centrifugation, IgG removal with Protein G Sepharose™, and HSA depletion with sodium chloride/ethanol precipitation. The protocol is streamlined to increase reproducibility and is compatible with many proteomie platforms, including two-dimensional gel electrophoresis, and high-performance liquid chromatography either offline or coupled online with a mass spectrometer. The reproducible depletion of lipids, IgG. and HSA permits a higher load of the remaining serum proteins, facilitating the identification of disease biomarkers.

Original languageEnglish (US)
Title of host publicationCardiovascular Proteomics
Subtitle of host publicationMethods and Protocols
EditorsFernando Vivanco
Pages365-371
Number of pages7
DOIs
StatePublished - Feb 5 2007

Publication series

NameMethods in Molecular Biology
Volume357
ISSN (Print)1064-3745

Keywords

  • Albumin
  • Biomarker
  • Depletion
  • Disease
  • HSA
  • IgG
  • Immunoglobulin
  • Lipid
  • Proteomics
  • Serum

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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