Advanced

Measuring Antibody Orientation at the Bacterial Surface

Shannon, Oonagh LU and Nordenfelt, Pontus LU (2017) In Bacterial Pathogenesis 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)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
in
Bacterial Pathogenesis
editor
Nordenfelt, Pontus; Collin, Matthias; and
volume
1535
pages
7 pages
publisher
Springer New York
external identifiers
  • scopus:85005976037
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
2018-01-07 11:50:50
@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},
  editor       = {Nordenfelt, Pontus and Collin, Matthias},
  isbn         = {978-1-4939-6671-4},
  issn         = {1064-3745},
  language     = {eng},
  pages        = {331--337},
  publisher    = {Springer New York},
  series       = {Bacterial Pathogenesis},
  title        = {Measuring Antibody Orientation at the Bacterial Surface},
  url          = {http://dx.doi.org/10.1007/978-1-4939-6673-8_22},
  volume       = {1535},
  year         = {2017},
}