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Pre-analytical sample handling effects on tear fluid protein levels

Tear fluid is emerging as a source of non-invasive biomarkers, both for ocular and systemic conditions. Accurate quantification of tear proteins can be improved by standardizing methods to collect and process tear fluid. The aim of this study was to determine sample handling factors that may influen...

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Autores principales: Gijs, Marlies, Arumugam, Sinthuja, van de Sande, Nienke, Webers, Carroll A. B., Sethu, Swaminathan, Ghosh, Arkasubhra, Shetty, Rohit, Vehof, Jelle, Nuijts, Rudy M. M. A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9873914/
https://www.ncbi.nlm.nih.gov/pubmed/36693949
http://dx.doi.org/10.1038/s41598-023-28363-z
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author Gijs, Marlies
Arumugam, Sinthuja
van de Sande, Nienke
Webers, Carroll A. B.
Sethu, Swaminathan
Ghosh, Arkasubhra
Shetty, Rohit
Vehof, Jelle
Nuijts, Rudy M. M. A.
author_facet Gijs, Marlies
Arumugam, Sinthuja
van de Sande, Nienke
Webers, Carroll A. B.
Sethu, Swaminathan
Ghosh, Arkasubhra
Shetty, Rohit
Vehof, Jelle
Nuijts, Rudy M. M. A.
author_sort Gijs, Marlies
collection PubMed
description Tear fluid is emerging as a source of non-invasive biomarkers, both for ocular and systemic conditions. Accurate quantification of tear proteins can be improved by standardizing methods to collect and process tear fluid. The aim of this study was to determine sample handling factors that may influence the tear protein biomarker profile. Tear fluid was collected using Schirmer’s strips. Tear proteins were extracted by elution through centrifugation. Total protein content was determined using the bicinchoninic acid assay. Key concepts that apply to the entire sample processing cycle are tear sampling, tear storage, protein extraction and data normalization. Differences in wetting or migration length were observed between Schirmer’s strips from different manufacturers, and between protein-free and protein-rich solutions. One unit of migration length (mm) did not correspond to one unit of volume (µL). A positive correlation (r = 0.6671, p < 0.0001) was observed between migration length and total tear protein content. The most beneficial storage conditions were strips that were not stored (+ 21.8%), or underwent ‘wet’ storage (+ 11.1%). Protein recovery was the highest in 400 µL extraction buffer and independent of protein molecular weight. This study helps to explain inter- and intra-variability that is often seen with tear biomarker research. This information is critical to ensure accuracy of test results, as tear biomarkers will be used for patient management and in clinical trials in the near future. This study also highlights the need for standardization of Schirmer’s strip manufacturing, tear fluid processing and analyte concentration normalization.
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spelling pubmed-98739142023-01-26 Pre-analytical sample handling effects on tear fluid protein levels Gijs, Marlies Arumugam, Sinthuja van de Sande, Nienke Webers, Carroll A. B. Sethu, Swaminathan Ghosh, Arkasubhra Shetty, Rohit Vehof, Jelle Nuijts, Rudy M. M. A. Sci Rep Article Tear fluid is emerging as a source of non-invasive biomarkers, both for ocular and systemic conditions. Accurate quantification of tear proteins can be improved by standardizing methods to collect and process tear fluid. The aim of this study was to determine sample handling factors that may influence the tear protein biomarker profile. Tear fluid was collected using Schirmer’s strips. Tear proteins were extracted by elution through centrifugation. Total protein content was determined using the bicinchoninic acid assay. Key concepts that apply to the entire sample processing cycle are tear sampling, tear storage, protein extraction and data normalization. Differences in wetting or migration length were observed between Schirmer’s strips from different manufacturers, and between protein-free and protein-rich solutions. One unit of migration length (mm) did not correspond to one unit of volume (µL). A positive correlation (r = 0.6671, p < 0.0001) was observed between migration length and total tear protein content. The most beneficial storage conditions were strips that were not stored (+ 21.8%), or underwent ‘wet’ storage (+ 11.1%). Protein recovery was the highest in 400 µL extraction buffer and independent of protein molecular weight. This study helps to explain inter- and intra-variability that is often seen with tear biomarker research. This information is critical to ensure accuracy of test results, as tear biomarkers will be used for patient management and in clinical trials in the near future. This study also highlights the need for standardization of Schirmer’s strip manufacturing, tear fluid processing and analyte concentration normalization. Nature Publishing Group UK 2023-01-24 /pmc/articles/PMC9873914/ /pubmed/36693949 http://dx.doi.org/10.1038/s41598-023-28363-z Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Gijs, Marlies
Arumugam, Sinthuja
van de Sande, Nienke
Webers, Carroll A. B.
Sethu, Swaminathan
Ghosh, Arkasubhra
Shetty, Rohit
Vehof, Jelle
Nuijts, Rudy M. M. A.
Pre-analytical sample handling effects on tear fluid protein levels
title Pre-analytical sample handling effects on tear fluid protein levels
title_full Pre-analytical sample handling effects on tear fluid protein levels
title_fullStr Pre-analytical sample handling effects on tear fluid protein levels
title_full_unstemmed Pre-analytical sample handling effects on tear fluid protein levels
title_short Pre-analytical sample handling effects on tear fluid protein levels
title_sort pre-analytical sample handling effects on tear fluid protein levels
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9873914/
https://www.ncbi.nlm.nih.gov/pubmed/36693949
http://dx.doi.org/10.1038/s41598-023-28363-z
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