title: Succinic anhydride functionalized microcantilevers for protein immobilization and quantification.
abstract: Microcantilever-based systems have been proposed as sensing platforms owing to their high sensitivity when used as mass sensors. The controlled immobilization on a surface of biomolecules used as recognition elements is fundamental in order to realize a highly specific and sensitive biosensor. Here, we introduce for the first time the application to a microcantilever-based system of a reliable chemical functionalization consisting of silanization with an aminosilane followed by a modification resulting in a carboxylated thin film. This chemical functionalization was tested for reproducibility of molecule deposition and for its protein grafting ability. Finally, this system was employed for the quantification of grafted proteins on the microcantilever surface. Moreover, a theoretical surface density of immobilized proteins estimated with bioinformatics tools was compared with the experimental surface density data, providing information about the orientation that the biomolecules assumed with respect to the sensing surface.
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129, Torino, Italy. gianluca.palmara@polito.it.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Palmara
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129, Torino, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Chiadò
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129, Torino, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Calmo
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129, Torino, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Ricciardi
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D019204:GTP-Binding Proteins / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry*; D008962:Models, Theoretical; D049329:Nanostructures / Q000737:chemistry*; D011437:Propylamines / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012821:Silanes / Q000737:chemistry*; D013387:Succinic Anhydrides / Q000737:chemistry*
chemical_list: D055767:Immobilized Proteins; D011437:Propylamines; D011994:Recombinant Proteins; D012821:Silanes; D013387:Succinic Anhydrides; D012710:Serum Albumin, Bovine; C031801:succinic anhydride; D019204:GTP-Binding Proteins; C477625:amino-propyl-triethoxysilane
keywords: Biosensor; Chemical functionalization; Microcantilever; Protein immobilization; Protein orientation; Surface density