Measuring Antibody Orientation at the Bacterial Surface
Shannon, Oonagh LU and Nordenfelt, Pontus LU
(2017)
In Methods in Molecular Biology
1535.
p.331-337
- Abstract
Many bacteria have the ability to interact with antibodies as a means to circumvent the immune response. This includes binding to the Fc portion of antibodies, effectively reversing the antibody orientation and thus decreasing the Fc-mediated immune signaling. Since antibody orientation at the bacterial surface has been shown to be important in human disease, it is valuable to be able to assess how antibodies are interacting with bacterial pathogens. Here, we describe a method to measure the proportion of human IgG that are bound via their Fc or Fabs to a bacterial surface. This is achieved by treating antibody-coated bacteria with the bacterial enzyme IdeS - which will cleave IgG into Fc and Fab fragments - and subsequently detect... (More)
Many bacteria have the ability to interact with antibodies as a means to circumvent the immune response. This includes binding to the Fc portion of antibodies, effectively reversing the antibody orientation and thus decreasing the Fc-mediated immune signaling. Since antibody orientation at the bacterial surface has been shown to be important in human disease, it is valuable to be able to assess how antibodies are interacting with bacterial pathogens. Here, we describe a method to measure the proportion of human IgG that are bound via their Fc or Fabs to a bacterial surface. This is achieved by treating antibody-coated bacteria with the bacterial enzyme IdeS - which will cleave IgG into Fc and Fab fragments - and subsequently detect remaining fragments with fluorescent Fabs. The method is easy and fast, and the principle is most likely also applicable to other systems where distinguishing between antibody Fc and Fab binding is important.
(Less)
- author
- Shannon, Oonagh
LU
and Nordenfelt, Pontus
LU
- organization
- publishing date
- 2017
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- host publication
- Bacterial Pathogenesis : Methods and Protocols - Methods and Protocols
- series title
- Methods in Molecular Biology
- editor
- Nordenfelt, Pontus and Collin, Matthias
- volume
- 1535
- pages
- 7 pages
- publisher
- Springer
- external identifiers
-
- scopus:85005976037
- pmid:27914090
- ISSN
- 1064-3745
- ISBN
- 978-1-4939-6671-4
- 978-1-4939-6673-8
- DOI
- 10.1007/978-1-4939-6673-8_22
- language
- English
- LU publication?
- yes
- id
- 98f16d5f-d0fa-4855-a1e7-3bf9c1ff8091
- date added to LUP
- 2017-02-16 08:02:48
- date last changed
- 2025-01-07 07:28:29
@inbook{98f16d5f-d0fa-4855-a1e7-3bf9c1ff8091,
abstract = {{<p>Many bacteria have the ability to interact with antibodies as a means to circumvent the immune response. This includes binding to the Fc portion of antibodies, effectively reversing the antibody orientation and thus decreasing the Fc-mediated immune signaling. Since antibody orientation at the bacterial surface has been shown to be important in human disease, it is valuable to be able to assess how antibodies are interacting with bacterial pathogens. Here, we describe a method to measure the proportion of human IgG that are bound via their Fc or Fabs to a bacterial surface. This is achieved by treating antibody-coated bacteria with the bacterial enzyme IdeS - which will cleave IgG into Fc and Fab fragments - and subsequently detect remaining fragments with fluorescent Fabs. The method is easy and fast, and the principle is most likely also applicable to other systems where distinguishing between antibody Fc and Fab binding is important.</p>}},
author = {{Shannon, Oonagh and Nordenfelt, Pontus}},
booktitle = {{Bacterial Pathogenesis : Methods and Protocols}},
editor = {{Nordenfelt, Pontus and Collin, Matthias}},
isbn = {{978-1-4939-6671-4}},
issn = {{1064-3745}},
language = {{eng}},
pages = {{331--337}},
publisher = {{Springer}},
series = {{Methods in Molecular Biology}},
title = {{Measuring Antibody Orientation at the Bacterial Surface}},
url = {{http://dx.doi.org/10.1007/978-1-4939-6673-8_22}},
doi = {{10.1007/978-1-4939-6673-8_22}},
volume = {{1535}},
year = {{2017}},
}