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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.
Affiliation
  • Joseph SR; The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia.
  • Lum B; The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia.
  • Banushi B; The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia.
  • Barry R; The University of Queensland Diamantina Institute, University of Queensland, Woolloongabba, QLD 4102, Australia.
  • Panizza B; Princess Alexandra Hospital, Brisbane, QLD 4102, Australia.
  • Walpole E; Faculty of Medicine, University of Queensland, Woolloongabba, QLD 4102, Australia.
  • Simpson F; Princess Alexandra Hospital, Brisbane, QLD 4102, Australia.
STAR Protoc ; 1(2): 100087, 2020 09 18.
Article in En | MEDLINE | ID: mdl-33111120
ABSTRACT
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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Full text: 1 Database: MEDLINE Main subject: Immunohistochemistry / ErbB Receptors / EGF Family of Proteins Type of study: Guideline / Prognostic_studies Limits: Animals / Humans Language: En Journal: STAR Protoc Year: 2020 Type: Article Affiliation country: Australia
Fulltext
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Full text: 1 Database: MEDLINE Main subject: Immunohistochemistry / ErbB Receptors / EGF Family of Proteins Type of study: Guideline / Prognostic_studies Limits: Animals / Humans Language: En Journal: STAR Protoc Year: 2020 Type: Article Affiliation country: Australia