April 11, 2024
Journal Article

A novel assay to measure low-density lipoproteins binding to proteoglycans


Background: The binding of low-density lipoprotein (LDL) to proteoglycans (PGs) in the extracellular matrix (ECM) of the arterial intima is a key initial step in the development of atherosclerosis. Although many techniques have been developed to assess this binding, most of the methods are labor-intensive and technically challenging to standardize across research laboratories. Thus, sensitive, and reproducible assay to detect LDL binding to PGs is needed to screen clinical populations for atherosclerosis risk. Objectives: The aim of this study was to develop a quantitative, and reproducible assay to evaluate the affinity of LDL towards PGs and to replicate previously published results on LDL-PG binding. Methods: Immunofluorescence microscopy was performed to visualize the binding of LDL to PGs using mouse vascular smooth muscle (MOVAS) cells. An in-cell ELISA (ICE) was also developed and optimized to quantitatively measure LDL-PG binding using fixed MOVAS cells cultured in a 96-well format. Results: We used the ICE assay to show that, despite equal APOB concentrations, LDL isolated from adults with cardiovascular disease bound to PG to a greater extent than LDL isolated from adults without cardiovascular disease (p

Published: April 11, 2024


Geh E., D.K. Swertfeger, H. Sexmith, A. Heink, P. Tarapore, J.T. Melchior, and W. Davidson, et al. 2024. A novel assay to measure low-density lipoproteins binding to proteoglycans. PLoS One 19, no. 1:Art. No. e0291632. PNNL-SA-190079. doi:10.1371/journal.pone.0291632

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