Analysis of intracellular trafficking of dendritic cell receptors for antigen targeting

Haiyin Liu, Claire Dumont, Angus P R Johnston, Justine D. Mintern

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

5 Citations (Scopus)

Abstract

Antibody-targeted vaccination aims to efficiently deliver antigen to dendritic cells by targeting specific receptors at the cell surface. The choice of receptor depends on different factors, including their capacity to induce internalization of the delivered antigen/adjuvant cargo. Assays currently used to monitor internalization in dendritic cells have several limitations. We have developed a novel DNA-based probe that allows for simple and robust high-throughput analysis of internalization. Designed for flow cytometry, the probe can also be used for fluorescence microscopy to clearly distinguish internalized from surface-bound material. Here, we describe the steps for modifying material (antibodies, proteins) with the probe, undertaking the assay, and analyzing the data obtained from flow cytometry.

Original languageEnglish
Title of host publicationDendritic Cell Protocols
EditorsElodie Segura, Nobuyuki Onai
Place of PublicationNew York NY USA
PublisherHumana Press
Chapter15
Pages199-209
Number of pages11
Volume1423
Edition3rd
ISBN (Electronic)9781493936069
ISBN (Print)9781493936045
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
PublisherSpringer Nature
Volume1423
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Antibodies
  • Binding
  • Dendritic cell
  • Endocytosis
  • Flow cytometry
  • Fluorescence
  • Internalization
  • Receptor
  • Sensor

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