UV-light-assisted functionalization for sensing of light molecules

Riccardo Funari; Bartolomeo Della Ventura; Antonio Ambrosio; Stefano Lettieri; Pasqualino Maddalena; Carlo Altucci; Raffaele Velotta

doi:10.1117/12.2018510

3 May 2013 UV-light-assisted functionalization for sensing of light molecules

Riccardo Funari, Bartolomeo Della Ventura, Antonio Ambrosio, Stefano Lettieri, Pasqualino Maddalena, Carlo Altucci, Raffaele Velotta

Author Affiliations +

Proceedings Volume 8774, Optical Sensors 2013; 87740K (2013) https://doi.org/10.1117/12.2018510
Event: SPIE Optics + Optoelectronics, 2013, Prague, Czech Republic

Abstract

An antibody immobilization technique based on the formation of thiol groups after UV irradiation of the proteins is shown to be able to orient upside antibodies on a gold electrode of a Quartz Crystal Microbalance (QCM). This greatly affects the aptitude of antibodies in recognizing small antigens thereby increasing the sensitivity of the QCM. The capability of such a procedure to orient antibodies is confirmed by the Atomic Force Microscopy (AFM) of the surface that shows different statistical distributions for the height of the detected peaks, whether the irradiation is performed or not. In particular, the distributions are Gaussian with a standard deviation smaller when irradiated antibodies are used compared to that obtained with no treated antibodies. The standard deviation reduction is explained in terms of higher order induced on the host surface resulting from the trend of irradiated antibodies to be anchored upside on the surface with their antigen binding sites free to catch recognized analytes. As a result the sensitivity of the realized biosensor is increased by even more than one order of magnitude.

Citation Download Citation

Riccardo Funari, Bartolomeo Della Ventura, Antonio Ambrosio, Stefano Lettieri, Pasqualino Maddalena, Carlo Altucci, and Raffaele Velotta "UV-light-assisted functionalization for sensing of light molecules", Proc. SPIE 8774, Optical Sensors 2013, 87740K (3 May 2013); https://doi.org/10.1117/12.2018510

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KEYWORDS

Proteins

Gold

Electrodes

Ultraviolet radiation

Luminescence

Molecules

Atomic force microscopy

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