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

Shannon R. Joseph; Benedict Lum; Blerida Banushi; Rachael Barry; Benedict Panizza; Euan Walpole; Fiona Simpson

STAR Protocols (Sep 2020)

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

Shannon R. Joseph,
Benedict Lum,
Blerida Banushi,
Rachael Barry,
Benedict Panizza,
Euan Walpole,
Fiona Simpson

Affiliations

Shannon R. Joseph: The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia
Benedict Lum: The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia
Blerida Banushi: The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia
Rachael Barry: The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia
Benedict Panizza: Princess Alexandra Hospital, Brisbane, QLD 4102, Australia; Faculty of Medicine, University of Queensland, Woolloongabba, QLD 4102, Australia
Euan Walpole: Princess Alexandra Hospital, Brisbane, QLD 4102, Australia; Faculty of Medicine, University of Queensland, Woolloongabba, QLD 4102, Australia
Fiona Simpson: The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia; Corresponding author

Journal volume & issue: Vol. 1, no. 2
p. 100087

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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