A monoclonal antibody to assess oxidized cholesteryl esters associated with apoAI and apoB-100 lipoproteins in human plasma

• Published in: Journal of lipid research Vol. 60; no. 2; pp. 436 - 445
• Main Authors: Gonen, Ayelet, Choi, Soo-Ho, Miu, Phuong, Agatisa-Boyle, Colin, Acks, Daniel, Taylor, Angela M., McNamara, Coleen A., Tsimikas, Sotirios, Witztum, Joseph L., Miller, Yury I.
• Format: Journal Article
• Language: English
• Published: United States The American Society for Biochemistry and Molecular Biology 01.02.2019
• Subjects: Animals; Antibodies, Monoclonal - immunology; Apolipoprotein A-I - metabolism; Apolipoprotein B-100 - metabolism; Cholesterol Esters - blood; Cholesterol Esters - immunology; Cholesterol Esters - metabolism; Enzyme-Linked Immunosorbent Assay - methods; Humans; Methods; Mice; Oxidation-Reduction; oxidation-specific epitope; apolipoprotein B-100; apolipoprotein AI; biomarker
• ISSN: 0022-2275; 1539-7262; 1539-7262
• DOI: 10.1194/jlr.D090852

Atherosclerosis is associated with increased lipid peroxidation, leading to generation of multiple oxidation-specific epitopes (OSEs), contributing to the pathogenesis of atherosclerosis and its clinical manifestation. Oxidized cholesteryl esters (OxCEs) are a major class of OSEs found in human plasma and atherosclerotic tissue. To evaluate OxCEs as a candidate biomarker, we generated a novel mouse monoclonal Ab (mAb) specific to an OxCE modification of proteins. The mAb AG23 (IgG1) was raised in C57BL6 mice immunized with OxCE-modified keyhole limpet hemocyanin, and hybridomas were screened against OxCE-modified BSA. This method ensures mAb specificity to the OxCE modification, independent of a carrier protein. AG23 specifically stained human carotid artery atherosclerotic lesions. An ELISA method, with AG23 as a capture and either anti-apoAI or anti-apoB-100 as the detection Abs, was developed to assay apoAI and apoB-100 lipoproteins that have one or more OxCE epitopes. OxCE-apoA or OxCE-apoB did not correlate with the well-established oxidized phospholipid-apoB biomarker. In a cohort of subjects treated with atorvastatin, OxCE-apoA was significantly lower than in the placebo group, independent of the apoAI levels. These results suggest the potential diagnostic utility of a new biomarker assay to measure OxCE-modified lipoproteins in patients with CVD.