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ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab

Circulating HER2 extracellular domain (HER2 ECD) levels were proposed as a surrogate for HER2 tissue expression to monitor breast cancer patients for early relapse or responses to standard or HER2-targeted therapies, such as the monoclonal antibody (mAb) trastuzumab. Currently, available commercial...

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Autores principales: Agnolon, Valentina, Contato, Anna, Meneghello, Anna, Tagliabue, Elda, Toffoli, Giuseppe, Gion, Massimo, Polo, Federico, Fabricio, Aline S. C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7033231/
https://www.ncbi.nlm.nih.gov/pubmed/32080226
http://dx.doi.org/10.1038/s41598-020-59630-y
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author Agnolon, Valentina
Contato, Anna
Meneghello, Anna
Tagliabue, Elda
Toffoli, Giuseppe
Gion, Massimo
Polo, Federico
Fabricio, Aline S. C.
author_facet Agnolon, Valentina
Contato, Anna
Meneghello, Anna
Tagliabue, Elda
Toffoli, Giuseppe
Gion, Massimo
Polo, Federico
Fabricio, Aline S. C.
author_sort Agnolon, Valentina
collection PubMed
description Circulating HER2 extracellular domain (HER2 ECD) levels were proposed as a surrogate for HER2 tissue expression to monitor breast cancer patients for early relapse or responses to standard or HER2-targeted therapies, such as the monoclonal antibody (mAb) trastuzumab. Currently, available commercial ELISA assays for HER2 ECD rely on antibodies recognizing undisclosed or unknown epitopes. In this work, two ELISA assays employing MGR2 and MGR3 epitope-specific mAbs for HER2 ECD were developed and validated, showing good assay precision and linearity of the dose-response signal within the dynamic range of 0.19–12.50 ng mL(−1) and detection limits of 0.76 and 0.75 ng mL(−1) for the MGR2 and MGR3 assays, respectively. The developed assay showed a good agreement with two widely used commercial kits for HER2 ECD quantification in serum samples from breast cancer patients. A complete characterization of mAb-HER2 ECD interaction was performed by means of surface plasmon resonance using trastuzumab as control for both epitope mapping and kinetics analysis. The epitopes recognized by the two mAbs showed no overlap with trastuzumab, which was confirmed by trastuzumab interference analysis in serum samples. The method showed to be a practical approach to determine HER2 ECD with a high degree of sensitivity, reliability and recovery in samples containing mAbs-based therapies.
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spelling pubmed-70332312020-02-28 ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab Agnolon, Valentina Contato, Anna Meneghello, Anna Tagliabue, Elda Toffoli, Giuseppe Gion, Massimo Polo, Federico Fabricio, Aline S. C. Sci Rep Article Circulating HER2 extracellular domain (HER2 ECD) levels were proposed as a surrogate for HER2 tissue expression to monitor breast cancer patients for early relapse or responses to standard or HER2-targeted therapies, such as the monoclonal antibody (mAb) trastuzumab. Currently, available commercial ELISA assays for HER2 ECD rely on antibodies recognizing undisclosed or unknown epitopes. In this work, two ELISA assays employing MGR2 and MGR3 epitope-specific mAbs for HER2 ECD were developed and validated, showing good assay precision and linearity of the dose-response signal within the dynamic range of 0.19–12.50 ng mL(−1) and detection limits of 0.76 and 0.75 ng mL(−1) for the MGR2 and MGR3 assays, respectively. The developed assay showed a good agreement with two widely used commercial kits for HER2 ECD quantification in serum samples from breast cancer patients. A complete characterization of mAb-HER2 ECD interaction was performed by means of surface plasmon resonance using trastuzumab as control for both epitope mapping and kinetics analysis. The epitopes recognized by the two mAbs showed no overlap with trastuzumab, which was confirmed by trastuzumab interference analysis in serum samples. The method showed to be a practical approach to determine HER2 ECD with a high degree of sensitivity, reliability and recovery in samples containing mAbs-based therapies. Nature Publishing Group UK 2020-02-20 /pmc/articles/PMC7033231/ /pubmed/32080226 http://dx.doi.org/10.1038/s41598-020-59630-y Text en © The Author(s) 2020 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Agnolon, Valentina
Contato, Anna
Meneghello, Anna
Tagliabue, Elda
Toffoli, Giuseppe
Gion, Massimo
Polo, Federico
Fabricio, Aline S. C.
ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
title ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
title_full ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
title_fullStr ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
title_full_unstemmed ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
title_short ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
title_sort elisa assay employing epitope-specific monoclonal antibodies to quantify circulating her2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7033231/
https://www.ncbi.nlm.nih.gov/pubmed/32080226
http://dx.doi.org/10.1038/s41598-020-59630-y
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