Measurement of Antibody Binding Affinity on Bacterial Surfaces Using Flow Cytometry

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Antibody binding to bacterial surfaces plays a crucial role in immunity, and a key characteristic of this protein-protein interaction is the binding affinity. Determining the affinity of an antibody binding to its antigen is the first step in predicting the function in a physiological environment where other competing protein interactions may be present. Antibody-antigen affinity is often evaluated with isolated proteins. It is informative to also be able to assess antibody binding to a bacterial surface where many antigens might be present, including multiple copies of the specific antigen the antibody recognizes, and in a context where the antigen might be in a more natural conformation. In this chapter, we present a flow cytometry-based assay to measure and calculate the cell surface binding affinity or avidity of any mono- or polyclonal antibody solution.

Original languageEnglish
Title of host publicationBacterial Pathogenesis
Subtitle of host publicationMethods and Protocols
EditorsPontus Nordenfelt, Mattias Collin
Place of PublicationNew York
Number of pages9
Publication statusPublished - 2023

Publication series

NameMethods in molecular biology (Clifton, N.J.)
ISSN (Print)1940-6029

Subject classification (UKÄ)

  • Immunology in the medical area
  • Biochemistry and Molecular Biology

Free keywords

  • Flow Cytometry
  • Antibody Affinity
  • Antigens
  • Antibodies


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