title: Surface Plasmon Resonance Method to Evaluate Anti-citrullinated Protein/Peptide Antibody Affinity to Citrullinated Peptides.
abstract: Surface plasmon resonance (SPR) technique is extremely interesting in immunology because it has the potential to directly visualize biomolecular interactions in real-time monitoring antibody affinity, one of the parameters affecting pathogenicity in autoimmune diseases. Herein we describe the affinity evaluation of anti-citrullinated peptide antibodies (ACPA) to a peptide-based biosensor by SPR. The method describes the purification of ACPA isolated from rheumatoid arthritis (RA) patients using affinity columns, the strategy employed for the immobilization of citrullinated peptides onto a sensor chip, and the evaluation of the specific binding of purified ACPA to immobilized peptides.
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Division of Pharmaceutical Sciences and Nutraceutic, Department of NeuroFarBa, University of Florence, Sesto Fiorentino, Italy.
authors.forename: Feliciana
authors.initials: F
authors.lastname: Real-Fernández
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Division of Pharmaceutical Sciences and Nutraceutic, Department of NeuroFarBa, University of Florence, Sesto Fiorentino, Italy.
authors.forename: Giada
authors.initials: G
authors.lastname: Rossi
authors.affiliation: Clinical Immunology and Allergy Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Filomena
authors.initials: F
authors.lastname: Panza
authors.affiliation: Clinical Immunology and Allergy Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Pratesi
authors.affiliation: Clinical Immunology and Allergy Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Migliorini
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Division of Pharmaceutical Sciences and Nutraceutic, Department of NeuroFarBa, University of Florence, Sesto Fiorentino, Italy. paolo.rovero@unifi.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Rovero
mesh_terms: D000906:Antibodies / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity / Q000276:immunology*; D002846:Chromatography, Affinity; D002956:Citrulline / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010455:Peptides / Q000737:chemistry* / Q000276:immunology*; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D000906:Antibodies; D010455:Peptides; D002956:Citrulline
keywords: Antibody affinity; Biacore; Citrullinated peptides; Kinetic; Surface plasmon resonance (SPR)