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Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
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Published on: October 15, 2016

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Epitope Mapping of Antibodies Using Bacterial Cell Surface Display of Gene Fragment Libraries

Anna-Luisa Volk1, Francis Jingxin Hu1, Johan Rockberg2

  • 1School of Biotechnology, AlbaNova University Center, KTH-Royal Institute of Technology, Stockholm, Sweden.

Methods in Molecular Biology (Clifton, N.J.)
|May 2, 2018
PubMed

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.

Keywords:
AntibodyCell surface displayEpitope mappingFACSGram-positiveS. carnosus

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