Optical Detection of Integrin AvB3 Using a Nanostructured Lipid Bilayer Membrane Biosensor
O. Worsfold and T. Nishiya
Fujirebio Inc., JP
Keywords: integrin, optical, bionanosensor, lipid membrane
An ultra-sensitive optical bionanosensor platform for integrin avb3 loaded human umbilical vein endothelial cells (HUVECs) was fabricated. The bionanosensor design involved the incorporation of an RGD (arginine-glycineaspartate) containing peptide conjugated BODIPY lipid soluble dye (donor) into a substrate supported bilayer lipid membrane (SBLM). The RGD peptide binds with the integrin avb3 via a multimerisation mechanism thus reducing the energy transfer from the donor dye moiety to a second higher excitation wavelength BODIPY lipid soluble dye (acceptor) moiety. The reduction in energy transfer is visualised by an increase in the fluorescence intensity of the donor and a decrease in the fluorescence intensity of the acceptor. The surface chemistry of the substrate has a significant effect on the fluidity of the lipid bilayer (measured by Fluorescence Recovery after Photobleaching (FRAP)) and on the subsequent biosensing response. Using this novel bionanosensor architecture a sensitivity of 25 HUVEC cells/ml was recorded.
Nanotech 2004 Conference Technical Program Abstract