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Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG

Bilirubin is a toxicological biomarker for hemolysis and liver diseases. The current automated diazo method used in clinical chemistry has limited applicability in rodent models and cannot be used in small animals relevant to toxicology, microphysiological systems, cell cultures, and kinetic studies...

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Detalles Bibliográficos
Autores principales: Sist, Paola, Tramer, Federica, Bandiera, Antonella, Urbani, Ranieri, Redenšek Trampuž, Sara, Dolžan, Vita, Passamonti, Sabina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10671013/
https://www.ncbi.nlm.nih.gov/pubmed/38003479
http://dx.doi.org/10.3390/ijms242216289
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author Sist, Paola
Tramer, Federica
Bandiera, Antonella
Urbani, Ranieri
Redenšek Trampuž, Sara
Dolžan, Vita
Passamonti, Sabina
author_facet Sist, Paola
Tramer, Federica
Bandiera, Antonella
Urbani, Ranieri
Redenšek Trampuž, Sara
Dolžan, Vita
Passamonti, Sabina
author_sort Sist, Paola
collection PubMed
description Bilirubin is a toxicological biomarker for hemolysis and liver diseases. The current automated diazo method used in clinical chemistry has limited applicability in rodent models and cannot be used in small animals relevant to toxicology, microphysiological systems, cell cultures, and kinetic studies. Here, we present a versatile fluorometric method for nanoscale analysis of bilirubin based on its highly specific binding to the recombinant bifunctional protein HELP–UnaG (HUG). The assay is sensitive (LoQ = 1.1 nM), accurate (4.5% relative standard error), and remarkably robust, allowing analysis at pH 7.4–9.5, T = 25–37 °C, in various buffers, and in the presence of 0.4–4 mg × L(−1) serum albumin or 30% DMSO. It allows repeated measurements of bilirubinemia in murine models and small animals, fostering the 3Rs principle. The assay determines bilirubin in human plasma with a relative standard error of 6.7% at values that correlate and agree with the standard diazo method. Furthermore, it detects differences in human bilirubinemia related to sex and UGT1A1 polymorphisms, thus demonstrating its suitability for the uniform assessment of bilirubin at the nanoscale in translational and precision medicine.
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spelling pubmed-106710132023-11-14 Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG Sist, Paola Tramer, Federica Bandiera, Antonella Urbani, Ranieri Redenšek Trampuž, Sara Dolžan, Vita Passamonti, Sabina Int J Mol Sci Article Bilirubin is a toxicological biomarker for hemolysis and liver diseases. The current automated diazo method used in clinical chemistry has limited applicability in rodent models and cannot be used in small animals relevant to toxicology, microphysiological systems, cell cultures, and kinetic studies. Here, we present a versatile fluorometric method for nanoscale analysis of bilirubin based on its highly specific binding to the recombinant bifunctional protein HELP–UnaG (HUG). The assay is sensitive (LoQ = 1.1 nM), accurate (4.5% relative standard error), and remarkably robust, allowing analysis at pH 7.4–9.5, T = 25–37 °C, in various buffers, and in the presence of 0.4–4 mg × L(−1) serum albumin or 30% DMSO. It allows repeated measurements of bilirubinemia in murine models and small animals, fostering the 3Rs principle. The assay determines bilirubin in human plasma with a relative standard error of 6.7% at values that correlate and agree with the standard diazo method. Furthermore, it detects differences in human bilirubinemia related to sex and UGT1A1 polymorphisms, thus demonstrating its suitability for the uniform assessment of bilirubin at the nanoscale in translational and precision medicine. MDPI 2023-11-14 /pmc/articles/PMC10671013/ /pubmed/38003479 http://dx.doi.org/10.3390/ijms242216289 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sist, Paola
Tramer, Federica
Bandiera, Antonella
Urbani, Ranieri
Redenšek Trampuž, Sara
Dolžan, Vita
Passamonti, Sabina
Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG
title Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG
title_full Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG
title_fullStr Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG
title_full_unstemmed Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG
title_short Nanoscale Bilirubin Analysis in Translational Research and Precision Medicine by the Recombinant Protein HUG
title_sort nanoscale bilirubin analysis in translational research and precision medicine by the recombinant protein hug
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10671013/
https://www.ncbi.nlm.nih.gov/pubmed/38003479
http://dx.doi.org/10.3390/ijms242216289
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