Any molecular recognition reaction supported by a solid-phase drives a specific change of the solid-solution interfacial tension. Sessile Contact Angle (CA) experiments can be readily used to track this thermodynamic parameter, prompting this well-known technique to be reinvented as an alternative, easy-access and label-free way to probe and study molecular recognition events. Here we deploy this technique, renamed for this application CONAMORE (CONtact Angle MOlecular REcognition), to study the interaction of the tumor-derived pro-angiogenic vascular endothelial growth factor-A (VEGF-A) with the extracellular domain of its receptor VEGFR2. We show that CONAMORE recognizes the high affinity binding of VEGF-A at nanomolar concentrations to surface-immobilized VEGFR2 regardless of the presence of a ten folds excess of a non specific interacting protein, and that it further proofs its specificity and reliability on competitive binding experiments involving neutralizing anti-VEGF-A antibodies. Finally, CONAMORE shows the outstanding capability to detect the specific interaction between VEGFR2 and low molecular weight ligands, such as Cyclo-VEGI, a VEGFR2 antagonist cyclo-peptide, that weights about 2 kDa.

Nanoliter contact angle probes tumor angiogenic ligand-receptor protein interactions

OLIVIERO, Giulio;MAIOLO, Daniele;LEALI, Daria;FEDERICI, Stefania;DEPERO, Laura Eleonora;PRESTA, Marco;MITOLA, Stefania Maria Filomena;BERGESE, Paolo
2010-01-01

Abstract

Any molecular recognition reaction supported by a solid-phase drives a specific change of the solid-solution interfacial tension. Sessile Contact Angle (CA) experiments can be readily used to track this thermodynamic parameter, prompting this well-known technique to be reinvented as an alternative, easy-access and label-free way to probe and study molecular recognition events. Here we deploy this technique, renamed for this application CONAMORE (CONtact Angle MOlecular REcognition), to study the interaction of the tumor-derived pro-angiogenic vascular endothelial growth factor-A (VEGF-A) with the extracellular domain of its receptor VEGFR2. We show that CONAMORE recognizes the high affinity binding of VEGF-A at nanomolar concentrations to surface-immobilized VEGFR2 regardless of the presence of a ten folds excess of a non specific interacting protein, and that it further proofs its specificity and reliability on competitive binding experiments involving neutralizing anti-VEGF-A antibodies. Finally, CONAMORE shows the outstanding capability to detect the specific interaction between VEGFR2 and low molecular weight ligands, such as Cyclo-VEGI, a VEGFR2 antagonist cyclo-peptide, that weights about 2 kDa.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11379/39628
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