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Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers

The expression level of the HER family is unreliable as a predictive marker for targeted therapies in cancer. Thus, there is a need to develop other biomarkers, which can be used to accurately select responsive patients for targeted therapies. The HER dimerization status may be more important than H...

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Detalles Bibliográficos
Autores principales: Waterhouse, Benjamin R., Gijsen, Merel, Barber, Paul R., Tullis, Iain D.C., Vojnovic, Borivoj, Kong, Anthony
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3248220/
https://www.ncbi.nlm.nih.gov/pubmed/21908901
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author Waterhouse, Benjamin R.
Gijsen, Merel
Barber, Paul R.
Tullis, Iain D.C.
Vojnovic, Borivoj
Kong, Anthony
author_facet Waterhouse, Benjamin R.
Gijsen, Merel
Barber, Paul R.
Tullis, Iain D.C.
Vojnovic, Borivoj
Kong, Anthony
author_sort Waterhouse, Benjamin R.
collection PubMed
description The expression level of the HER family is unreliable as a predictive marker for targeted therapies in cancer. Thus, there is a need to develop other biomarkers, which can be used to accurately select responsive patients for targeted therapies. The HER dimerization status may be more important than HER receptor expression per se in determining sensitivity or resistance to a given therapeutic agent. The aim of the study is to develop a FRET assay using dye conjugated secondary antibodies to assess HER receptor dimerization. Using primary antibodies from different species in conjunction with Alexa488 and Alexa546 conjugated secondary antibodies, we validated our EGFR/HER2 dimerization assay in three cell lines, EGFR positive A431 cells as well as HER2 positive breast cell lines BT474 and SKBR3 cells. Finally, we applied our assay to assess EGFR/HER2 dimerization in paraffin embedded cell pellets. Our results show promise for the assay to be applied to tumor samples in order to assess the prognostic significance and predictive value of HER receptor dimerization in various cancers.
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spelling pubmed-32482202012-01-18 Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers Waterhouse, Benjamin R. Gijsen, Merel Barber, Paul R. Tullis, Iain D.C. Vojnovic, Borivoj Kong, Anthony Oncotarget Methodological Reports The expression level of the HER family is unreliable as a predictive marker for targeted therapies in cancer. Thus, there is a need to develop other biomarkers, which can be used to accurately select responsive patients for targeted therapies. The HER dimerization status may be more important than HER receptor expression per se in determining sensitivity or resistance to a given therapeutic agent. The aim of the study is to develop a FRET assay using dye conjugated secondary antibodies to assess HER receptor dimerization. Using primary antibodies from different species in conjunction with Alexa488 and Alexa546 conjugated secondary antibodies, we validated our EGFR/HER2 dimerization assay in three cell lines, EGFR positive A431 cells as well as HER2 positive breast cell lines BT474 and SKBR3 cells. Finally, we applied our assay to assess EGFR/HER2 dimerization in paraffin embedded cell pellets. Our results show promise for the assay to be applied to tumor samples in order to assess the prognostic significance and predictive value of HER receptor dimerization in various cancers. Impact Journals LLC 2011-09-08 /pmc/articles/PMC3248220/ /pubmed/21908901 Text en Copyright: © 2011 Waterhouse et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
spellingShingle Methodological Reports
Waterhouse, Benjamin R.
Gijsen, Merel
Barber, Paul R.
Tullis, Iain D.C.
Vojnovic, Borivoj
Kong, Anthony
Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
title Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
title_full Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
title_fullStr Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
title_full_unstemmed Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
title_short Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
title_sort assessment of egfr/her2 dimerization by fret-flim utilizing alexa-conjugated secondary antibodies in relation to targeted therapies in cancers
topic Methodological Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3248220/
https://www.ncbi.nlm.nih.gov/pubmed/21908901
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