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

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.

doi:10.1194/jlr.d090852

Cited by 7 publications

(7 citation statements)

References 42 publications

(49 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The OxCE used for covalent modification of these proteins was a product of cholesteryl arachidonate oxidation with 2,2'-azobis (2,4-dimethylvaleronitrile) in an atmosphere of oxygen, which predominantly produced BEP-CE, but other complex oxidation products were present as well (44). The AG23 immunoreactivity was abundant in human carotid endarterectomy specimens, demonstrating the presence of OxCE epitopes in atherosclerotic lesions (Figure 1-Biomarkers) and suggesting relevance of OxCE to the pathogenesis of human CVD (61).…”

Section: Oxce In Human Atherosclerotic Lesionsmentioning

confidence: 99%

“…To ensure the independence of an OxCE epitope recognition from the protein carrier, mice were immunized with OxCE-KLH and the antibody was selected against OxCE-BSA. The resulting antibody, AG23, bound OxCE-modified KLH, BSA, apoA-I, and a 6-amino acid peptide (61). The OxCE used for covalent modification of these proteins was a product of cholesteryl arachidonate oxidation with 2,2'-azobis (2,4-dimethylvaleronitrile) in an atmosphere of oxygen, which predominantly produced BEP-CE, but other complex oxidation products were present as well (44).…”

Section: Oxce In Human Atherosclerotic Lesionsmentioning

confidence: 99%

“…The AG23 mAb against OxCE described in the preceding section has been used to develop a new ELISA method to measure OxCE associated with apoA-I or apoB-100 lipoproteins in human plasma (61). This assay measures levels of lipoproteins that have at least one OxCE epitope.…”

Section: Oxce In Human Plasma As a Biomarker For Cvdmentioning

confidence: 99%

See 2 more Smart Citations

From Inert Storage to Biological Activity—In Search of Identity for Oxidized Cholesteryl Esters

Gonen

Miller

2020

Front. Endocrinol.

Self Cite

View full text Add to dashboard Cite

Esterification of cholesterol is a universal mechanism to store and transport large quantities of cholesterol between organs and tissues and to avoid toxicity of the excess of cellular cholesterol. Intended for transport and storage and thus to be inert, cholesteryl esters (CEs) reside in hydrophobic cores of circulating lipoproteins and intracellular lipid droplets. However, the inert identity of CEs is dramatically changed if cholesterol is esterified to a polyunsaturated fatty acid and subjected to oxidative modification. Post-synthetic, or epilipidomic, oxidative modifications of CEs are mediated by specialized enzymes, chief among them are lipoxygenases, and by free radical oxidation. The complex repertoire of oxidized CE (OxCE) products exhibit various, context-dependent biological activities, surveyed in this review. Oxidized fatty acyl chains in OxCE can be hydrolyzed and re-esterified, thus seeding oxidized moieties into phospholipids (PLs), with OxPLs having different from OxCEs biological activities. Technological advances in mass spectrometry and the development of new anti-OxCE antibodies make it possible to validate the presence and quantify the levels of OxCEs in human atherosclerotic lesions and plasma. The article discusses the prospects of measuring OxCE levels in plasma as a novel biomarker assay to evaluate risk of developing cardiovascular disease and efficacy of treatment.

show abstract

Section: Oxce In Human Atherosclerotic Lesionsmentioning

confidence: 99%

Section: Oxce In Human Atherosclerotic Lesionsmentioning

confidence: 99%

See 1 more Smart Citation

From Inert Storage to Biological Activity—In Search of Identity for Oxidized Cholesteryl Esters

Gonen

Miller

2020

Front. Endocrinol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…A BALB/C mouse was immunized with a similar protocol as previously described (25), with a primary boost of 125 g of the 8K-IV apo(a) construct, followed at 2 week intervals with three intraperitoneal boosts with 75 g. A final antigen boost of 25 g of 8K-IV in PBS was administered intravenously 2 weeks after the third boost.…”

Section: Immunization Of Mice and Generation And Purification Of Monoclonal Antibodiesmentioning

confidence: 99%

“…Titrations of antibodies present in immunized mouse plasma, screening of hybridoma-conditioned media, and testing the purified monoclonal antibody were performed by chemiluminescent ELISA as previously described (25). In brief, 96-well microtiter plates (Brand, Germany) were coated directly with Lp(a), apo(a) KIV 2 construct, or plasminogen at 5 g/ml in PBS overnight at 4°C.…”

Section: Elisa With Direct Antigen Plating To Assess Binding Of Hybridoma-generated Antibodiesmentioning

confidence: 99%

Generation and characterization of LPA-KIV9, a murine monoclonal antibody binding a single site on apolipoprotein (a)

Gonen

Yang

Yeang

et al. 2020

Journal of Lipid Research

Self Cite

View full text Add to dashboard Cite

Lipoprotein(a) [Lp(a)] is a risk factor for cardiovascular disease and a target of therapy, but Lp(a) measurements are not globally standardized. Commercially-available assays generally use polyclonal antibodies that detect multiple sites within the kringle (K) IV2 repeat region of Lp(a) and may lead to inaccurate assessments of plasma levels. With increasing awareness of Lp(a) as a cardiovascular risk factor and the active clinical development of new potential therapeutic approaches, the broad availability of reagents capable of providing isoform-independence of Lp(a) measurements is paramount. To address this issue, we generated a murine monoclonal antibody that binds to only one site on apolipoprotein(a). A BALB/C mouse was immunized with a truncated version of apolipoprotein(a) that contained eight total KIV repeats, including only one copy of KIV2. We generated hybridomas, screened them and successfully produced a KIV2-independent monoclonal antibody, named LPA-KIV9. Using a variety of truncated apolipoprotein(a) constructs to map its binding site, we found that LPA-KIV9 binds to KIV9, without binding to plasminogen. Fine peptide mapping revealed that LPA-KIV9 bound to the sequence 4076LETPTVV4082 on KIV9. In conclusion, the generation of monoclonal antibody LPA-KIV9 may be a useful reagent in basic research studies and in the clinical application of Lp(a) measurements.

show abstract

An update on lipid oxidation and inflammation in cardiovascular diseases

Zhong

Shen

et al. 2019

Free Radical Biology and Medicine

257

156

View full text Add to dashboard Cite

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

Cited by 7 publications

References 42 publications

From Inert Storage to Biological Activity—In Search of Identity for Oxidized Cholesteryl Esters

From Inert Storage to Biological Activity—In Search of Identity for Oxidized Cholesteryl Esters

Generation and characterization of LPA-KIV9, a murine monoclonal antibody binding a single site on apolipoprotein (a)

An update on lipid oxidation and inflammation in cardiovascular diseases

Contact Info

Product

Resources

About