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Immunofixation Electrophoresis of Serum Apolipoproteins A-I, A-II, and B, and Comparison with Cholesterol Fractions
Conference poster   Peer reviewed

Immunofixation Electrophoresis of Serum Apolipoproteins A-I, A-II, and B, and Comparison with Cholesterol Fractions

Rachel C James, F N Cornell and Mark A Holmes
Clinical Biochemist Reviews, Vol.25(Supplement), p.S102
10th Asian Pacific Congress of Clinical Biochemistry and the 42nd Australasian Association of Clinical Biochemists' Annual Scientific Conference, 2004 (Perth, Australia, 19-Sep-2004–23-Sep-2004)
Australasian Association of Clinical Biochemists
2004
url
http://www.aacb.asn.au/professionaldevelopment/the-clinical-biochemist-reviewsView
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Abstract

Medical Biochemistry and Metabolomics immunofixation electrophoresis cholesterol apolipoproteins
Introduction: Apolipoproteins (apo) A-I, A-II and B are important emerging biochemical markers of cardiovascular disease risk. The aim of this study was to develop an immunofixation electrophoresis (IFE) method for the analysis of serum apo A-I, A-II and B, and compare the staining patterns to cholesterol fractions. Methods: Venous blood was collected from 10 healthy adults following an overnight fast. Serum lipoprotein fractions were separated by agarose gel electrophoresis using a REP® VIS cholesterol kit (Helena Laboratories) for both quantitative cholesterol determination and apolipoprotein immunofixation. The apolipoproteins were located on the gels using goat polyclonal antisera (Chemicon), followed by staining. Optimum concentrations of serum and antisera were established. Results: IFE of serum apo A-1 resulted in a large and small band in the alpha region (HDL), and a faint unidentified band in the beta region. The apo A-II staining pattern was similar to that observed for apo A-I. However, the small, more cathodic apo A-I band in the alpha region did not react with apo A-II antisera. IFE of erum apo B resulted in a large slower moving band in the beta region (LDL), smaller bands in the pre-beta (VLDL) and fast pre-beta (Lpa; when elevated), and a small unidentified band in the alpha region. The location and density of the apolipoprotein bands were compared with the cholesterol profile. Discussion: The IFE method described is suitable for the qualitative and semi-quantitative analysis of serum apo A-I, A-II and B, and is sufficiently sensitive to detect apolipoprotein variations between individuals, with comparison to cholesterol fractions. Further refinement is required before the IFE method can be used quantitatively. Poster presented at the 10th Asian Pacific Congress of Clinical Biochemistry in conjunction with the AACB's 42nd Annual Scientific Conference.

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