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Nanoscale bilirubin analysis in translational research and precision medicine by the recombinant protein HUG [Elektronski vir]Sist, Paola, kemija ...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.Source: International journal of molecular sciences [Elektronski vir]. - ISSN 1422-0067 (Vol. 24, iss. 22, 2023, str. 1-24)Type of material - e-articlePublish date - 2023Language - englishCOBISS.SI-ID - 173321475
Author
Sist, Paola, kemija |
Tramer, Federica |
Bandiera, Antonella |
Urbani, Ranieri |
Redenšek Trampuž, Sara |
Dolžan, Vita |
Passamonti, Sabina
Topics
bilirubin |
fluorescent method |
calibration |
high-throughput assay |
biomedical diagnostics |
biomarker |
bilirubin |
fluorescentna metoda |
kalibracija |
test visoke zmogljivosti |
biomedicinska diagnostika |
biološki označevalec
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Sist, Paola, kemija | ![]() |
Tramer, Federica | ![]() |
Bandiera, Antonella | ![]() |
Urbani, Ranieri | ![]() |
Redenšek Trampuž, Sara | 38239 |
Dolžan, Vita | 11711 |
Passamonti, Sabina | ![]() |
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