Multiplex epitope mapping using bacterial surface display reveals both linear and conformational epitopes
2012 (English)In: Scientific Reports, ISSN 2045-2322, Vol. 2, 706- p.Article in journal (Refereed) Published
As antibody-based diagnosis and therapy grow at an increased pace, there is a need for methods which rapidly and accurately determine antibody-antigen interactions. Here, we report a method for the multiplex determination of antibody epitopes using bacterial cell-surface display. A protein-fragment library with 107 cell clones, covering 60 clinically-relevant protein targets, was created and characterized with massively parallel sequencing. Using this multi-target fragment library we determined simultaneously epitopes of commercial monoclonal and polyclonal antibodies targeting PSMA, EGFR, and VEGF. Off-target binding was observed for one of the antibodies, which demonstrates the method's ability to reveal cross-reactivity. We exemplify the detection of structural epitopes by mapping the therapeutic antibody Avastin. Based on our findings we suggest this method to be suitable for mapping linear and structural epitopes of monoclonal and polyclonal antibodies in a multiplex fashion and could find applicability in serum profiling as well as other protein-protein interaction studies.
Place, publisher, year, edition, pages
2012. Vol. 2, 706- p.
Growth-Factor Receptor, Cell-Surface, Staphylococcus-Carnosus, Recombinant Proteins, Crystal-Structure, Flow-Cytometry, Binding Site, Antibodies, Fab, Antigen
IdentifiersURN: urn:nbn:se:kth:diva-104361DOI: 10.1038/srep00706ISI: 000309524900004ScopusID: 2-s2.0-84874551743OAI: oai:DiVA.org:kth-104361DiVA: diva2:564789
FunderScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201211052012-11-052012-11-012012-11-05Bibliographically approved