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Measurement of Antibody Binding Affinity on Bacterial Surfaces Using Flow Cytometry

Kumra Ahnlide, Vibha LU and Nordenfelt, Pontus LU orcid (2023) In Methods in molecular biology (Clifton, N.J.) p.251-259
Abstract

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... (More)

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.

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Please use this url to cite or link to this publication:
author
and
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Flow Cytometry, Antibody Affinity, Antigens, Antibodies
host publication
Bacterial Pathogenesis : Methods and Protocols - Methods and Protocols
series title
Methods in molecular biology (Clifton, N.J.)
editor
Nordenfelt, Pontus and Collin, Mattias
edition
2
pages
9 pages
external identifiers
  • pmid:37258973
  • scopus:85160790177
ISSN
1940-6029
ISBN
978-1-0716-3242-0
DOI
10.1007/978-1-0716-3243-7_17
language
English
LU publication?
yes
additional info
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
id
292e7844-c305-4233-b263-83f04251ec8d
date added to LUP
2023-06-09 14:16:24
date last changed
2025-04-20 09:08:24
@inbook{292e7844-c305-4233-b263-83f04251ec8d,
  abstract     = {{<p>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.</p>}},
  author       = {{Kumra Ahnlide, Vibha and Nordenfelt, Pontus}},
  booktitle    = {{Bacterial Pathogenesis : Methods and Protocols}},
  editor       = {{Nordenfelt, Pontus and Collin, Mattias}},
  isbn         = {{978-1-0716-3242-0}},
  issn         = {{1940-6029}},
  keywords     = {{Flow Cytometry; Antibody Affinity; Antigens; Antibodies}},
  language     = {{eng}},
  pages        = {{251--259}},
  series       = {{Methods in molecular biology (Clifton, N.J.)}},
  title        = {{Measurement of Antibody Binding Affinity on Bacterial Surfaces Using Flow Cytometry}},
  url          = {{http://dx.doi.org/10.1007/978-1-0716-3243-7_17}},
  doi          = {{10.1007/978-1-0716-3243-7_17}},
  year         = {{2023}},
}