A gel-based method for purification of apolipoprotein A-I from small volumes of plasma

Brace, Rachel J., Sorrenson, Brie, Sviridov, Dmitri and McCormick, Sally P. A. (2010) A gel-based method for purification of apolipoprotein A-I from small volumes of plasma. Journal of Lipid Research, 51 11: 3370-3376. doi:10.1194/jlr.D008300

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Author Brace, Rachel J.
Sorrenson, Brie
Sviridov, Dmitri
McCormick, Sally P. A.
Title A gel-based method for purification of apolipoprotein A-I from small volumes of plasma
Journal name Journal of Lipid Research   Check publisher's open access policy
ISSN 0022-2275
Publication date 2010-11-01
Sub-type Article (original research)
DOI 10.1194/jlr.D008300
Open Access Status File (Publisher version)
Volume 51
Issue 11
Start page 3370
End page 3376
Total pages 7
Place of publication Rockville, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Subject 1303 Biochemistry
1310 Endocrinology
1307 Cell Biology
Abstract We present here a gel-based method for rapid purification of apolipoprotein A-I (apoA-I) from small volumes of human plasma. After isolation of high density lipoprotein from plasma, the apoA-I protein was separated by electrophoresis and the apoA-I band excised from the gel. The apoA-I was then eluted from the gel strip, concentrated, and delipidated ready for use. The structure and function of the gel-purified apoA-I protein was compared against apoA-I purified by the traditional size-exclusion chromatography method. The α-helical content of the gel-purified apoA-I as determined by circular dichroism was similar to chromatography-purified apoA-I. The functional activity of gel-purified apoA-I, as determined by cholesterol efflux assays in primary human fibroblasts and RAW264.7 macrophages, was also comparable with chromatography-purified apoA-I. This method is a valid alternative for apoA-I purification with some advantages over traditional chromatography purification including a much reduced plasma volume requirement, less time and cost, and a higher percentage protein recovery. The method is particularly suitable for applications requiring the purification of apoA-I from multiple human or animal samples of interest.
Keyword Cholesterol efflux
Gel purification
High density lipoproteins
Size-exclusion chromatography
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Queensland Brain Institute Publications
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Created: Mon, 30 Nov 2015, 21:55:13 EST by Rachel Suetani on behalf of Queensland Brain Institute