Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health

Abstract Respiratory health of children is a health priority. Club cell protein (CC16) is an interesting biomarker of lung diseases and adverse effects towards the airway epithelium integrity. Osteopontin (OPN) and nuclear factor-kappa B (NF-κB) also play a role in respiratory health. The use of uri...

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Autores principales:	Sarah J. D. Nauwelaerts, Nancy H. C. Roosens, Alfred Bernard, Sigrid C. J. De Keersmaecker, Koen De Cremer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/73f24a3249654d2798c6178373c628d8
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spelling	oai:doaj.org-article:73f24a3249654d2798c6178373c628d82021-12-02T16:50:27ZDevelopment of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health10.1038/s41598-021-89068-92045-2322https://doaj.org/article/73f24a3249654d2798c6178373c628d82021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89068-9https://doaj.org/toc/2045-2322Abstract Respiratory health of children is a health priority. Club cell protein (CC16) is an interesting biomarker of lung diseases and adverse effects towards the airway epithelium integrity. Osteopontin (OPN) and nuclear factor-kappa B (NF-κB) also play a role in respiratory health. The use of urine as biomarker source is useful in studies involving children but necessitates proper adjustment for physiological confounders influencing the urinary excretion, potentially characterized with beta-2-microglobulin (β2M), retinol binding protein 4 (RBP4) or myoglobin (MYO), as well as adjustment for possible renal dysfunction, characterized by human serum albumin (HSA). The simultaneous quantification of all these proteins in urine could facilitate children’s health monitoring. A multiple reaction monitoring method (MRM) was developed and validated for the relative quantification of the seven mentioned urinary proteins. A total of nine proteotypic peptides were selected and used for the relative quantification of the seven proteins. The MRM method was completely validated for all proteins and partially for OPN. LOQ’s ranged from 0.3 to 42.8 ng/ml, a good reproducibility and a good linearity were obtained across the analytical measurement range (r2 > 0.98). The method yielded varying correlations (r2 of 0.78, 0.71, 0.34 and 0.15 for CC16, β2M, RBP4 and HSA respectively) with available immunoassay data. It also allowed the identification and successful quantification of β2M and RBP4 as a protein candidate for adjustment of renal handling and dysfunction. All proteins were detected in the urine samples except for MYO and NF-κB. Our validated MRM-method is able to simultaneously quantify in urine biomarkers of airway epithelium integrity and biomarkers of variation in renal function and urinary dilution. This will allow to investigate further in future studies if urine can be used as a good surrogate source for biomarkers of airway epithelium integrity, and to understand the complex relationship between cause and effect in children’s respiratory health monitoring.Sarah J. D. NauwelaertsNancy H. C. RoosensAlfred BernardSigrid C. J. De KeersmaeckerKoen De CremerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sarah J. D. Nauwelaerts Nancy H. C. Roosens Alfred Bernard Sigrid C. J. De Keersmaecker Koen De Cremer Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
description	Abstract Respiratory health of children is a health priority. Club cell protein (CC16) is an interesting biomarker of lung diseases and adverse effects towards the airway epithelium integrity. Osteopontin (OPN) and nuclear factor-kappa B (NF-κB) also play a role in respiratory health. The use of urine as biomarker source is useful in studies involving children but necessitates proper adjustment for physiological confounders influencing the urinary excretion, potentially characterized with beta-2-microglobulin (β2M), retinol binding protein 4 (RBP4) or myoglobin (MYO), as well as adjustment for possible renal dysfunction, characterized by human serum albumin (HSA). The simultaneous quantification of all these proteins in urine could facilitate children’s health monitoring. A multiple reaction monitoring method (MRM) was developed and validated for the relative quantification of the seven mentioned urinary proteins. A total of nine proteotypic peptides were selected and used for the relative quantification of the seven proteins. The MRM method was completely validated for all proteins and partially for OPN. LOQ’s ranged from 0.3 to 42.8 ng/ml, a good reproducibility and a good linearity were obtained across the analytical measurement range (r2 > 0.98). The method yielded varying correlations (r2 of 0.78, 0.71, 0.34 and 0.15 for CC16, β2M, RBP4 and HSA respectively) with available immunoassay data. It also allowed the identification and successful quantification of β2M and RBP4 as a protein candidate for adjustment of renal handling and dysfunction. All proteins were detected in the urine samples except for MYO and NF-κB. Our validated MRM-method is able to simultaneously quantify in urine biomarkers of airway epithelium integrity and biomarkers of variation in renal function and urinary dilution. This will allow to investigate further in future studies if urine can be used as a good surrogate source for biomarkers of airway epithelium integrity, and to understand the complex relationship between cause and effect in children’s respiratory health monitoring.
format	article
author	Sarah J. D. Nauwelaerts Nancy H. C. Roosens Alfred Bernard Sigrid C. J. De Keersmaecker Koen De Cremer
author_facet	Sarah J. D. Nauwelaerts Nancy H. C. Roosens Alfred Bernard Sigrid C. J. De Keersmaecker Koen De Cremer
author_sort	Sarah J. D. Nauwelaerts
title	Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
title_short	Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
title_full	Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
title_fullStr	Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
title_full_unstemmed	Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
title_sort	development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/73f24a3249654d2798c6178373c628d8
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Development of a multiplex mass spectrometry method for simultaneous quantification of urinary proteins related to respiratory health

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