Enhancing Antibodies’ Binding Capacity through Oriented Functionalization of Plasmonic Surfaces

Coluccio, Maria Laura; Grillo, Fabiana; Onesto, Valentina; Garo, Virginia; Scala, Cinzia; Cuzzola, Paola; Calfa, Michela; Candeloro, Patrizio; Gentile, Francesco; Piletsky, Sergey; Malara, Natalia

Protein A has long been used in different research fields due to its ability to specifically recognize immunoglobulins (Ig). The protein derived from Staphylococcus aureus binds Ig through the Fc region of the antibody, showing its strongest binding in immunoglobulin G (IgG), making it the most used protein in its purification and detection. The research presented here integrates, for the first time, protein A to a silicon surface patterned with gold nanoparticles for the oriented binding of IgG. The signal detection is conveyed through a metal enhanced fluorescence (MEF) system. Orienting immunoglobulins allows the exposition of the fragment antigen-binding (Fab) region for the binding to its antigen, substantially increasing the binding capacity per antibody immobilized. Antibodies orientation is of crucial importance in many diagnostics devices, particularly when either component is in limited quantities.