title: Disposable amperometric magnetoimmunosensor for the sensitive detection of the cardiac biomarker amino-terminal pro-B-type natriuretic peptide in human serum.
abstract: A novel amperometric magnetoimmunosensor using an indirect competitive format is developed for the sensitive detection of the amino-terminal pro-B-type natriuretic peptide (NT-proBNP). The immunosensor design involves the covalent immobilization of the antigen onto carboxylic-modified magnetic beads (HOOC-MBs) activated with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), and further incubation in a mixture solution containing variable concentrations of the antigen and a fixed concentration of an HRP-labeled detection antibody. Accordingly, the target NT-proBNP in the sample and that immobilized on the MBs compete for binding to a fixed amount of the specific HRP-labeled secondary antibody. The immunoconjugate-bearing MBs are captured by a magnet placed under the surface of a disposable gold screen-printed electrode (Au/SPE). The amperometric responses measured at -0.10 V (vs. a Ag pseudo-reference electrode), upon addition of 3,3',5,5'-tetramethylbenzidine (TMB) as electron transfer mediator and H2O2 as the enzyme substrate, are used to monitor the affinity reaction. The developed magnetoimmunosensor provides attractive analytical characteristics in 10-times diluted human serum samples, exhibiting a linear range of clinical usefulness (0.12-42.9 ng mL(-1)) and a detection limit of 0.02 ng mL(-1), which can be used in clinical diagnosis of chronic heart failure in the elderly and for classifying patients at risk of death after heart transplantation. The magnetoimmunosensor was successfully applied to the analysis of spiked human serum samples.
authors.affiliation: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, Madrid, Spain. berta.efa@quim.ucm.es
authors.forename: Berta Esteban-Fernández
authors.initials: BE
authors.lastname: de Ávila
authors.forename: Vanessa
authors.initials: V
authors.lastname: Escamilla-Gómez
authors.forename: Susana
authors.initials: S
authors.lastname: Campuzano
authors.forename: María
authors.initials: M
authors.lastname: Pedrero
authors.forename: José M
authors.initials: JM
authors.lastname: Pingarrón
mesh_terms: D015415:Biomarkers / Q000097:blood; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods; D006333:Heart Failure / Q000097:blood*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D057230:Limit of Detection; D059346:Magnets; D020097:Natriuretic Peptide, Brain / Q000097:blood*; D010446:Peptide Fragments / Q000097:blood*; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D010446:Peptide Fragments; C109794:pro-brain natriuretic peptide (1-76); D020097:Natriuretic Peptide, Brain