Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples

We describe an ex vivo EGF ligand internalization assay using fresh patient tumor biopsies to determine how antigen targets will be trafficked before patients receive mAb treatment. This protocol facilitates a sensitive and reproducible indication as to mAbs surface retention times during treatment....

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Detalles Bibliográficos
Autores principales: Joseph, Shannon R., Lum, Benedict, Banushi, Blerida, Barry, Rachael, Panizza, Benedict, Walpole, Euan, Simpson, Fiona
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580087/
https://www.ncbi.nlm.nih.gov/pubmed/33111120
http://dx.doi.org/10.1016/j.xpro.2020.100087
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author Joseph, Shannon R.
Lum, Benedict
Banushi, Blerida
Barry, Rachael
Panizza, Benedict
Walpole, Euan
Simpson, Fiona
author_facet Joseph, Shannon R.
Lum, Benedict
Banushi, Blerida
Barry, Rachael
Panizza, Benedict
Walpole, Euan
Simpson, Fiona
author_sort Joseph, Shannon R.
collection PubMed
description We describe an ex vivo EGF ligand internalization assay using fresh patient tumor biopsies to determine how antigen targets will be trafficked before patients receive mAb treatment. This protocol facilitates a sensitive and reproducible indication as to mAbs surface retention times during treatment. EGF uptake protocols can also be used to analyze EGFR heterogeneity and localization of EGFR in both tumor and xenograft tissue. The technology can be adapted to analyze other receptors such as PD-L1 for which methods are provided. For complete details on the use and execution of this protocol, please refer to Joseph et al. (2019) and Chew et al. (2020).
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spelling pubmed-75800872020-10-26 Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples Joseph, Shannon R. Lum, Benedict Banushi, Blerida Barry, Rachael Panizza, Benedict Walpole, Euan Simpson, Fiona STAR Protoc Protocol We describe an ex vivo EGF ligand internalization assay using fresh patient tumor biopsies to determine how antigen targets will be trafficked before patients receive mAb treatment. This protocol facilitates a sensitive and reproducible indication as to mAbs surface retention times during treatment. EGF uptake protocols can also be used to analyze EGFR heterogeneity and localization of EGFR in both tumor and xenograft tissue. The technology can be adapted to analyze other receptors such as PD-L1 for which methods are provided. For complete details on the use and execution of this protocol, please refer to Joseph et al. (2019) and Chew et al. (2020). Elsevier 2020-08-13 /pmc/articles/PMC7580087/ /pubmed/33111120 http://dx.doi.org/10.1016/j.xpro.2020.100087 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Joseph, Shannon R.
Lum, Benedict
Banushi, Blerida
Barry, Rachael
Panizza, Benedict
Walpole, Euan
Simpson, Fiona
Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples
title Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples
title_full Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples
title_fullStr Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples
title_full_unstemmed Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples
title_short Antibody/Ligand-Target Receptor Internalization Assay Protocol Using Fresh Human or Murine Tumor Ex Vivo Samples
title_sort antibody/ligand-target receptor internalization assay protocol using fresh human or murine tumor ex vivo samples
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580087/
https://www.ncbi.nlm.nih.gov/pubmed/33111120
http://dx.doi.org/10.1016/j.xpro.2020.100087
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