Search: id:"swepub:oai:DiVA.org:kth-238351" >
Discontinuous epito...
Discontinuous epitope mapping of antibodies using bacterial cell surface display of folded domains
-
- Volk, Anna-Luisa (author)
- KTH,Skolan för bioteknologi (BIO)
-
- Rockberg, Johan (author)
- KTH,Proteinteknologi
-
(creator_code:org_t)
- 2018-05-01
- 2018
- English.
-
In: Epitope Mapping Protocols. - New York, NY : Humana Press. - 9781493978397 ; , s. 159-183
- Related links:
-
https://urn.kb.se/re...
-
show more...
-
https://doi.org/10.1...
-
show less...
Abstract
Subject headings
Close
- Knowledge of the exquisite-binding surface of an antibody on its target protein is of great value, in particular for therapeutic antibodies for understanding method of action and for stratification of patients carrying the necessary epitope for desired drug efficacy, but also for capture assays under native conditions. Several epitope mapping methodologies have been described for this purpose, with the laborious X-ray crystallography method being the ideal method for mapping of discontinuous epitopes in antibody-antigen crystal complexes and high-throughput peptide-based methods for mapping of linear epitopes. We here report on the usage of a bacterial surface display-based method for mapping of structural epitopes by display of folded domains on the surface of Gram positive bacteria, followed by domain-targeted mutagenesis and library analysis for the identification of key-residues by flow sorting and sequencing. Identified clones with reduced affinity are validated by single clone FACS and subsequent full-length expression in mammalian cells for validation.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology (hsv//eng)
Keyword
- Antibody
- Cell surface display
- CHO
- Discontinuous epitope mapping
- FACS
- Gram-positive
- S. carnosus
Publication and Content Type
- ref (subject category)
- kap (subject category)
Find in a library
To the university's database