Clinical Chemistry
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Clinical Chemistry 49: 1524-1526, 2003; 10.1373/49.9.1524
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(Clinical Chemistry. 2003;49:1524-1526.)
© 2003 American Association for Clinical Chemistry, Inc.

Technical Briefs

Antibody Phenotyping Test for the Human Apolipoprotein E2 Isoform

Robert L. Raffaï2,a, Ruth McPherson1, Karl H. Weisgraber2, Thomas L. Innerarity2, Eric Rassart3, Thomas P. Bersot2 and Ross W. Milne1

1 Lipoprotein and Atherosclerosis Group, University of Ottawa Heart Institute, Ottawa Civic Hospital, Ottawa, Ontario, K1Y 4E9 Canada

2 The Gladstone Institute of Cardiovascular Disease, Cardiovascular Research Institute, University of California, San Francisco, CA 94141-9100

3 Département des Sciences Biologiques, Université du Québec, Montréal, CP 8888, Québec, H3C 3P8 Canada

aauthor for correspondence: fax 415-285-5632, e-mail rraffai@gladstone.ucsf.edu

The first 20% of the full text of this article appears below.

Numerous methods have been described to determine the apolipoprotein E (apoE) phenotype or genotype of individuals (1)(2). These techniques are relatively time-consuming, and interpretation of the results can be difficult. Here, we report the development of a rapid and specific antibody-based test for the identification of the apoE2 isoform. Previously, we characterized the binding properties of a panel of anti-human apoE monoclonal antibodies (mAbs) (3)(4)(5). The locations of epitopes of selected mAbs from the panel are presented in Fig. 1A . Although neither mAb 6C5 nor 3H1 showed apoE isoform specificity, mAb 2E8 recognized an epitope that includes the apoE LDL receptor-binding site and resembles the LDL receptor in terms of its fine specificity.


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  		Figure 1. Immunometric assay for the E2 phenotypic isoform of human apoE.

(A), Epitope map of human apoE. Sandwich immunometric assays for human apoE were performed using mAb 6C5 as capture antibody, 3H1 for quantification, and 2E8 for determination of the apoE2 phenotypic isoform. (B and C), results obtained with the immunometric assay using the RIA version of the test, which is capable of unambiguously identifying the presence of the apoE2 isoform in homozygous as well as heterozygous plasma samples. (D), immunoreactivity of mAb 2E8 in mixtures of pure apoE2 and apoE3 isoforms, using the ELISA version of the test. The ratios in the legend are the . . . [Full Text of this Article]






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