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Methods Mol Biol. 2007;357:365-71.

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

Author information

1
Department of Medicine, Johns Hopkins University, Baltimore, MD, USA.

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 proteomic 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.

PMID:
17172702
DOI:
10.1385/1-59745-214-9:365
[Indexed for MEDLINE]

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