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Fully automated immunoassay for cholesterol uptake capacity to assess high-density lipoprotein function and cardiovascular disease risk

Murakami, Katsuhiro Harada, Amane Toh, Ryuji Kubo, Takuya Miwa, Keiko Kim, Jeeeun Kiriyama, Maria Iino, Takuya Nishikawa, Youichi Uno, Shin-Nosuke Akatsuchi, Kohei Nagao, Manabu Ishida, Tatsuro Hirata, Ken-ichi 神戸大学

2023.02.02

概要

High-density lipoprotein (HDL) cholesterol efflux capacity (CEC), which is a conventional metric of HDL function, has been associated with coronary heart disease risk. However, the CEC assay requires cultured cells and takes several days to perform. We previously established a cell-free assay to evaluate cholesterol uptake capacity (CUC) as a novel measure of HDL functionality and demonstrated its utility in coronary risk stratification. To apply this concept clinically, we developed a rapid and sensitive assay system based on a chemiluminescent magnetic particle immunoassay. The system is fully automated, providing high reproducibility. Measurement of CUC in serum is completed within 20 min per sample without HDL isolation, a notably higher throughput than that of the conventional CEC assay. CUC decreased with myeloperoxidase-mediated oxidation of HDL or in the presence of N-ethylmaleimide, an inhibitor of lecithin: cholesterol acyltransferase (LCAT), whereas CUC was enhanced by the addition of recombinant LCAT. Furthermore, CUC correlated with CEC even after being normalized by ApoA1 concentration and was significantly associated with the requirement for revascularization due to the recurrence of coronary lesions. Therefore, our new assay system shows potential for the accurate measurement of CUC in serum and permits assessing cardiovascular health.

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Author contributions

A.H., R.T., and K.H. conceived the project. Ka.M., A.H., and R.T. designed the study concept and wrote the

manuscript. T.K., S.U., and Y.N. designed and synthesized Bio-PEG3-cholesterol. Ka.M., Ke.M., J.K., M.K., T.I.,

and K.A. established assays for CUC. R.T., M.N., and T.I. collected samples and interpreted the clinical data. All

authors read and approved the final version of the manuscript.

Competing interests The authors declare no competing interests.

Additional information

Supplementary Information The online version contains supplementary material available at https://​doi.​org/​

10.​1038/​s41598-​023-​28953-x.

Correspondence and requests for materials should be addressed to A.H. or R.T.

Reprints and permissions information is available at www.nature.com/reprints.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and

institutional affiliations.

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