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A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients

Osteopontin (OPN) is a secreted glycophosphoprotein that derives its name from its high abundance in bone and secretion by osteoblasts. It is also secreted by a number of immune cells and, therefore, is present in human plasma at nanogram per millilitre levels where it affects cell adhesion and moti...

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Detalles Bibliográficos
Autores principales: Leslie, Andrew, Teh, Evelyn, Druker, Arik, Pinto, Devanand M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10309610/
https://www.ncbi.nlm.nih.gov/pubmed/37384615
http://dx.doi.org/10.1371/journal.pone.0281491
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author Leslie, Andrew
Teh, Evelyn
Druker, Arik
Pinto, Devanand M.
author_facet Leslie, Andrew
Teh, Evelyn
Druker, Arik
Pinto, Devanand M.
author_sort Leslie, Andrew
collection PubMed
description Osteopontin (OPN) is a secreted glycophosphoprotein that derives its name from its high abundance in bone and secretion by osteoblasts. It is also secreted by a number of immune cells and, therefore, is present in human plasma at nanogram per millilitre levels where it affects cell adhesion and motility. OPN is involved in several normal physiological processes; however, OPN dyregulation leads to overexpression by tumor cells leading to immune evasion and increased metastasis. Plasma OPN is primarily measured by enzyme-linked immunosorbent assay (ELISA). However, due to the complexity of the various OPN isoforms, conflicting results have been obtained on the use of OPN as a biomarker even in the same disease condition. These discrepant results may result from the difficulty in comparing ELISA results obtained with different antibodies that target unique OPN epitopes. Mass spectrometry can be used to quantify proteins in plasma and, by targeting OPN regions that do not bear post-translational modifications, may provide more consistent quantification. However, the low (ng/mL) levels in plasma present a significant analytical challenge. In order to develop a sensitive assay for plasma OPN, we explored a single-step precipitation method using a recently developed spin-tube format. Quantification was performed using isotope-dilution mass spectrometry. The concentration detection limit of this assay was 39 ± 15 ng/mL. The assay was applied to the analysis of plasma OPN in metastatic breast cancer patients, where levels from 17 to 53 ng/mL were detected. The sensitivity of the method is higher than previously published methods and sufficient for OPN detection in large, high grade tumors but still requires improvement in sensitivity to be widely applicable.
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spelling pubmed-103096102023-06-30 A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients Leslie, Andrew Teh, Evelyn Druker, Arik Pinto, Devanand M. PLoS One Research Article Osteopontin (OPN) is a secreted glycophosphoprotein that derives its name from its high abundance in bone and secretion by osteoblasts. It is also secreted by a number of immune cells and, therefore, is present in human plasma at nanogram per millilitre levels where it affects cell adhesion and motility. OPN is involved in several normal physiological processes; however, OPN dyregulation leads to overexpression by tumor cells leading to immune evasion and increased metastasis. Plasma OPN is primarily measured by enzyme-linked immunosorbent assay (ELISA). However, due to the complexity of the various OPN isoforms, conflicting results have been obtained on the use of OPN as a biomarker even in the same disease condition. These discrepant results may result from the difficulty in comparing ELISA results obtained with different antibodies that target unique OPN epitopes. Mass spectrometry can be used to quantify proteins in plasma and, by targeting OPN regions that do not bear post-translational modifications, may provide more consistent quantification. However, the low (ng/mL) levels in plasma present a significant analytical challenge. In order to develop a sensitive assay for plasma OPN, we explored a single-step precipitation method using a recently developed spin-tube format. Quantification was performed using isotope-dilution mass spectrometry. The concentration detection limit of this assay was 39 ± 15 ng/mL. The assay was applied to the analysis of plasma OPN in metastatic breast cancer patients, where levels from 17 to 53 ng/mL were detected. The sensitivity of the method is higher than previously published methods and sufficient for OPN detection in large, high grade tumors but still requires improvement in sensitivity to be widely applicable. Public Library of Science 2023-06-29 /pmc/articles/PMC10309610/ /pubmed/37384615 http://dx.doi.org/10.1371/journal.pone.0281491 Text en © 2023 Leslie et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Leslie, Andrew
Teh, Evelyn
Druker, Arik
Pinto, Devanand M.
A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
title A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
title_full A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
title_fullStr A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
title_full_unstemmed A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
title_short A targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
title_sort targeted isotope dilution mass spectrometry assay for osteopontin quantification in plasma of metastatic breast cancer patients
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10309610/
https://www.ncbi.nlm.nih.gov/pubmed/37384615
http://dx.doi.org/10.1371/journal.pone.0281491
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