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Epitope mapping of antibodies using bacterial cell surface display of gene fragment libraries
KTH, School of Biotechnology (BIO).ORCID iD: 0000-0003-1096-9061
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH).ORCID iD: 0000-0002-1389-5371
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH).ORCID iD: 0000-0002-9977-5724
2018 (English)In: Epitope Mapping Protocols, Humana Press, 2018, p. 141-157Chapter in book (Refereed)
Abstract [en]

The unique property of specific high affinity binding to more or less any target of interest has made antibodies tremendously useful in numerous applications. Hence, knowledge of the precise binding site (epitope) of antibodies on the target protein is one of the most important features for understanding its performance and determining its reliability in immunoassays. Here, we describe an updated protocol for high-resolution method for mapping epitopes of antibodies based on bacterial surface expression of antigen fragments followed by antibody-based flow cytometric analysis. Epitopes are determined by DNA sequencing of the sorted antibody-binding cells followed by sequence alignment back to the antigen sequence. The method described here has been useful for the mapping of both monoclonal and polyclonal antibodies with varying sizes of epitopes.

Place, publisher, year, edition, pages
Humana Press, 2018. p. 141-157
Series
Methods in Molecular Biology, ISSN 1064-3745 ; 1785
Keywords [en]
Antibody, Cell surface display, Epitope mapping, FACS, Gram-positive, S. carnosus
National Category
Other Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-238341DOI: 10.1007/978-1-4939-7841-0_10Scopus ID: 2-s2.0-85046366274OAI: oai:DiVA.org:kth-238341DiVA, id: diva2:1263304
Funder
Knut and Alice Wallenberg Foundation
Note

QC 20181115

Available from: 2018-11-15 Created: 2018-11-15 Last updated: 2018-11-15Bibliographically approved

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Volk, Anna-LuisaHu, Francis JingxinRockberg, Johan

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CiteExportLink to record
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Citation style
  • apa
  • harvard1
  • ieee
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  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
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  • Other locale
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Output format
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