Chapter title |
ORFeome Phage Display.
|
---|---|
Chapter number | 27 |
Book title |
Phage Display
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7447-4_27 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7446-7, 978-1-4939-7447-4
|
Authors |
Zantow, Jonas, Moreira, Gustavo Marçal Schmidt Garcia, Dübel, Stefan, Hust, Michael, Jonas Zantow, Gustavo Marçal Schmidt Garcia Moreira, Stefan Dübel, Michael Hust |
Abstract |
ORFeome phage display allows the efficient functional screening of entire proteomes or even metaproteomes to identify immunogenic proteins. For this purpose, randomly fragmented, whole genomes or metagenomes are cloned into a phage-display vector allowing positive selection for open reading frames (ORF) to improve the library quality. These libraries display all possible proteins encoded by a pathogen or a microbiome on the phage surface. Consequently, immunogenic proteins can be selected from these libraries using disease-related immunoglobulins from patient serum. ORFeome phage display in particular allows the identification of immunogenic proteins that are only expressed in the host-pathogen interaction but not in cultivation, as well as the detection of very low expressed and very small immunogens and immunogenic proteins of non-cultivable organisms. The identified immunogenic proteins are potential biomarkers for the development of diagnostic assays or vaccines. These articles will give an introduction to ORFeome phage-display technology and give detailed protocols to identify immunogenic proteins by phage display. |
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