title: Surface plasmon resonance-based methodology for anti-adalimumab antibody identification and kinetic characterization.
abstract: Adalimumab (ADA) is a TNF-α blocker drug antibody fully humanized and thus indistinguishable in structure and function from natural human IgG1, used in the juvenile idiopathic arthritis (JIA) treatment. Immunogenicity against the drug has been frequently detected in treated patients, and the presence of anti-ADA antibodies is correlated to treatment failure or lower clinical remission. Herein, we measured by surface plasmon resonance (SPR) both the binding and the affinity of anti-ADA antibodies to the ADA-immobilized biosensor. The binding of anti-ADA antibodies was evaluated by testing sera from ADA-treated patients (n = 30), untreated patients (n = 9), and healthy donors (n = 20) in the SPR biosensor. The optimal cut-off point was defined using the receiver operating characteristic curve (ROC-curve) analysis with 79 % (60.28 to 92.01 %, 95 % CI) sensitivity, 99 % (88.06 to 100.0 %, 95 % CI) specificity, and a positive likelihood ratio of 23. The area under the curve was 0.9298 (p < 0.0001). The apparent affinity of anti-ADA antibodies from pediatric patients' sera was measured, analyzing the interaction of anti-drug antibodies using whole sera, enriched IgG fractions, and isolated anti-ADA antibodies. The immobilized drug ADA interacted with purified antibodies at low affinities (10(-6) M > K D > 10(-9) M). Graphical Abstract Adalimumab immobilized on the biosensor chip surface detects specific anti-drug antibodies in treated patients' sera.
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Florence, Italy.|Department of Neurosciences, Psychology, Drug Research and Child Health - Section of Pharmaceutical Sciences and Nutraceutics, University of Florence, Via Ugo Schiff 6, 50019, Sesto Fiorentino, Italy.
authors.forename: Feliciana
authors.initials: F
authors.lastname: Real-Fernández
authors.affiliation: Pediatric Rheumatology, AOU Meyer, Viale Gaetano Pieraccini, 24, 50141, Florence, Italy.|Department of Neurosciences, Psychology, Drug Research and Child Health - Sezione Età Evolutiva, University of Florence, Viale Pieraccini 24, 50139, Florence, Italy.
authors.forename: Rolando
authors.initials: R
authors.lastname: Cimaz
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Florence, Italy.|Department of Neurosciences, Psychology, Drug Research and Child Health - Section of Pharmaceutical Sciences and Nutraceutics, University of Florence, Via Ugo Schiff 6, 50019, Sesto Fiorentino, Italy.
authors.forename: Giada
authors.initials: G
authors.lastname: Rossi
authors.affiliation: Pediatric Rheumatology, AOU Meyer, Viale Gaetano Pieraccini, 24, 50141, Florence, Italy.|Department of Neurosciences, Psychology, Drug Research and Child Health - Sezione Età Evolutiva, University of Florence, Viale Pieraccini 24, 50139, Florence, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Simonini
authors.affiliation: Pediatric Rheumatology, AOU Meyer, Viale Gaetano Pieraccini, 24, 50141, Florence, Italy.
authors.forename: Teresa
authors.initials: T
authors.lastname: Giani
authors.affiliation: Pediatric Rheumatology, AOU Meyer, Viale Gaetano Pieraccini, 24, 50141, Florence, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Pagnini
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Florence, Italy.|Department of Chemistry "Ugo Schiff", University of Florence, Via della Lastruccia 13, 50019, Sesto Fiorentino, Italy.
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Papini
authors.affiliation: Laboratory of Peptide and Protein Chemistry and Biology, Florence, Italy. paolo.rovero@unifi.it.|Department of Neurosciences, Psychology, Drug Research and Child Health - Section of Pharmaceutical Sciences and Nutraceutics, University of Florence, Via Ugo Schiff 6, 50019, Sesto Fiorentino, Italy. paolo.rovero@unifi.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Rovero
mesh_terms: D000068879:Adalimumab / Q000276:immunology*; D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007700:Kinetics; D008297:Male; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D000068879:Adalimumab
keywords: Adalimumab; Affinity; Anti-drug antibodies; Surface plasmon resonance