The Binding of EGFR to GM1(3) Hosted in Lipid Raft-Like Biomembranes Insighted by Plasmonic Resonance Techniques

We exploit Au/SiO2 plasmonic structures to check the effective binding activity of GM1(3) gangliosides hosted in physiological-like biomembranes, in presence of the Epidermal Growth Factor Receptor (EGFR). To this aim, we used bilayers that support the propagation of optical surface plasmon modes (plasmonic transducers, PTs) or guided modes (Plasmon Waveguide Resonators, PWRs). First, we measured the binding of EGFR to GM1(3) by using PTs. Indeed, effective interactions were evidenced, but with faint signals that prevented resolving dissociation kinetics. In order to enhance the optical responses, we turned our attention to PWRs. We first refined the design of a previously adopted Au/SiO2 PWR, finding that the nominal sensitivity is independent on SiO2 thickness but strongly dependent on its residual losses, due typically to a nonoptimal deposition process. We fabricated an improved Au/SiO2 resonator and tested the predicted signal enhancement by monitoring the binding of EGFR to GM3-enriched biomembranes. The measured signal was ~12-fold higher than that one measured using a PT, close to the maximum theoretical enhancement. The higher PWR response enabled us to detect the dissociation of EGFR from GM3, and the value of the apparent dissociation constant of the GM3-EGFR complex could be obtained.