title: Impedance-derived electrochemical capacitance spectroscopy for the evaluation of lectin-glycoprotein binding affinity.
abstract: Characterization of lectin-carbohydrate binding using label-free methods such as impedance-derived electrochemical capacitance spectroscopy (ECS) is desirable to evaluate specific interactions, for example, ArtinM lectin and horseradish peroxidase (HRP) glycoprotein, used here as a model for protein-carbohydrate binding affinity. An electroactive molecular film comprising alkyl ferrocene as a redox probe and ArtinM as a carbohydrate receptive center to target HRP was successfully used to determine the binding affinity between ArtinM and HRP. The redox capacitance, a transducer signal associated with the alkyl ferrocene centers, was obtained by ECS and used in the Langmuir adsorption model to obtain the affinity constant (1.6±0.6)×10(8) L mol(-1). The results shown herein suggest the feasibility of ECS application for lectin glycoarray characterization.
authors.affiliation: Department of Physical Chemistry, Chemistry Institute, São Paulo State University (Unesp), 55 Prof. Francisco Degni Street, ZIP Code: 14800-060, São Paulo, Brazil.
authors.forename: Adriano
authors.initials: A
authors.lastname: Santos
authors.affiliation: Department of Physical Chemistry, Chemistry Institute, São Paulo State University (Unesp), 55 Prof. Francisco Degni Street, ZIP Code: 14800-060, São Paulo, Brazil.
authors.forename: Fernanda C
authors.initials: FC
authors.lastname: Carvalho
authors.affiliation: Department of Cellular and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School. University of São Paulo (USP), 3900 Bandeirantes Avenue. ZIP Code: 14049-900, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Maria-Cristina
authors.initials: MC
authors.lastname: Roque-Barreira
authors.affiliation: Department of Physical Chemistry, Chemistry Institute, São Paulo State University (Unesp), 55 Prof. Francisco Degni Street, ZIP Code: 14800-060, São Paulo, Brazil. Electronic address: prbueno@iq.unesp.br.
authors.forename: Paulo R
authors.initials: PR
authors.lastname: Bueno
mesh_terms: D031622:Artocarpus / Q000737:chemistry; D015374:Biosensing Techniques; D058266:Dielectric Spectroscopy; D017097:Electric Impedance; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000032:analysis / Q000378:metabolism*; D055767:Immobilized Proteins / Q000302:isolation & purification / Q000378:metabolism; D037102:Lectins / Q000302:isolation & purification / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding
chemical_list: D006023:Glycoproteins; D055767:Immobilized Proteins; D037102:Lectins; D006735:Horseradish Peroxidase
keywords: ArtinM; Binding affinity constant; HRP; Impedance-derived electrochemical capacitance spectroscopy; Langmuir isotherm