| Chapter title |
Measuring Antibody Orientation at the Bacterial Surface.
|
|---|---|
| Chapter number | 22 |
| Book title |
Bacterial Pathogenesis
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6673-8_22 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6671-4, 978-1-4939-6673-8
|
| Authors |
Oonagh Shannon, Pontus Nordenfelt |
| Editors |
Pontus Nordenfelt, Mattias Collin |
| 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 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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Professor > Associate Professor | 1 | 17% |
| Student > Bachelor | 1 | 17% |
| Lecturer | 1 | 17% |
| Student > Doctoral Student | 1 | 17% |
| Unknown | 2 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 2 | 33% |
| Immunology and Microbiology | 1 | 17% |
| Biochemistry, Genetics and Molecular Biology | 1 | 17% |
| Unknown | 2 | 33% |