A conformation-based phage-display panning to screen neutralizing anti-VEGF VHHs with VEGFR2 mimicry behavior

International Journal of Biological Macromolecules
S Shirin ShahangianReyhaneh Sariri

Abstract

The potency of VEGF-based anti-angiogenic strategies in cancer therapy and the brilliant characteristics of VHHs motivated us to directly block VEGF binding to its receptor with neutralizing single domain antibodies, thereby fading away the VEGF signaling pathway. Considering with high resolution crystal structure of VEGF-RBD/VEGFR2 complex, we could adopt a combinatorial screening strategy: stringent panning and competition ELISA, to direct the panning procedure to dominantly screen the favorable binders that bind and block the key functional regions of VEGF. Based on competition assay, the majority of the screened clones (82%) showed the VEGFR2 mimicry behavior for binding to VEGF molecule. The phage pool gets enriched in favor of sequences that bind the receptor binding sites of VEGF. Different immunoassays and molecular docking simulation verified that all selected VHHs could bind and cover the receptor binding sites of VEGF. Consequently, some modifications in panning procedure with considering the structural features and detailed information of functional regions of a protein antigen, led us to successfully trap the high-affinity specific binders against its hot functional regions. Since the selected VHHs could cover the ...Continue Reading

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Citations

Sep 3, 2020·Analytical and Bioanalytical Chemistry·Zahra S RezaeiReza H Sajedi
Mar 19, 2020·PeerJ·Akhila Melarkode VattekatteAlexandre G de Brevern
Feb 25, 2021·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Seyede Atefeh HosseiniRoghaye Arezumand
May 1, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sedigheh Bagheri-ZiariKamran Pooshang Bagheri

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