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Analysis of vitamin D metabolites in biological samples using a nanoluc-based vitamin D receptor ligand sensing system: NLucVDR. | LitMetric

Analysis of vitamin D metabolites in biological samples using a nanoluc-based vitamin D receptor ligand sensing system: NLucVDR.

J Steroid Biochem Mol Biol

Department of Pharmaceutical Engineering, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan. Electronic address:

Published: October 2023

AI Article Synopsis

  • - The study evaluated a new assay system, NLucVDR, designed to measure vitamin D metabolites in serum and urine, using samples from 23 participants.
  • - Results showed strong correlations between the NLucVDR assay and standard methods for measuring 25(OH)D levels, particularly in serum, indicating the assay's reliability as a measurement tool.
  • - Interestingly, while the assay indicated much higher 25(OH)D levels in urine compared to standard methods, it still demonstrated a positive correlation with serum levels, suggesting it could help predict vitamin D status from urine samples.

Article Abstract

Many assays are currently being developed to measure the levels of vitamin D metabolites in various samples (such as blood, urine, and saliva). This study focused on the measurement of vitamin D metabolites in serum and urine using the NLucVDR assay system, which consists of a split-type nanoluciferase and ligand-binding domain (LBD) of the human vitamin D receptor. Blood and urine samples were collected from 23 participants to validate the NLucVDR assay. The 25(OH)D levels in the serum and urine determined by the NLucVDR assay showed good correlations with those determined by standard analytical methods (ECLIA for serum and LC-MS/MS for urine), with correlation coefficients of 0.923 and 0.844 for serum and urine samples, respectively. In the case of serum samples, 25(OH)D levels determined by the NLucVDR assay were in good agreement with those determined by ECLIA. Therefore, the NLucVDR assay is a useful tool for measuring serum 25(OH)D levels. The contribution of each vitamin D metabolite to the luminescence intensity obtained during the NLucVDR assay depends on its concentration and affinity for NLucVDR. Thus, the contribution of 25(OH)D in serum appears to be much higher than that of the other metabolites. In contrast, the 25(OH)D levels in the urine determined by the NLucVDR assay were more than 20-fold higher than those determined by a standard analytical method (LC-MS/MS), suggesting that some vitamin D metabolite(s) in the urine remarkably increased the luminescence intensity of the NLucVDR assay. Notably, the 25(OH)D concentration in the urine determined by the NLucVDR assay and the serum 25(OH)D concentration determined by standard analytical methods showed a significant positive correlation (r = 0.568). These results suggest that the analysis of a small amount of urine using the NLucVDR assay may be useful for predicting the serum 25(OH)D levels.

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Source
http://dx.doi.org/10.1016/j.jsbmb.2023.106367DOI Listing

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