title: β-Conglutin dual aptamers binding distinct aptatopes.
abstract: An aptamer was previously selected against the anaphylactic allergen β-conglutin (β-CBA I), which was subsequently truncated to an 11-mer and the affinity improved by two orders of magnitude. The work reported here details the selection and characterisation of a second aptamer (β-CBA II) selected against a second aptatope on the β-conglutin target. The affinity of this second aptamer was similar to that of the 11-mer, and its affinity was confirmed by three different techniques at three independent laboratories. This β-CBA II aptamer in combination with the previously selected β-CBA I was then exploited to a dual-aptamer approach. The specific and simultaneous binding of the dual aptamer (β-CBA I and β-CBA II) to different sites of β-conglutin was confirmed using both microscale thermophoresis and surface plasmon resonance where β-CBA II serves as the primary capturing aptamer and β-CBA I or the truncated β-CBA I (11-mer) as the secondary signalling aptamer, which can be further exploited in enzyme-linked aptamer assays and aptasensors.
authors.affiliation: Nanobiotechnology and Bioanalysis Group, Department of Chemical Engineering, Universitat Rovira I Virgili, 43007, Tarragona, Spain.
authors.forename: Miriam
authors.initials: M
authors.lastname: Jauset Rubio
authors.affiliation: Nanobiotechnology and Bioanalysis Group, Department of Chemical Engineering, Universitat Rovira I Virgili, 43007, Tarragona, Spain.
authors.forename: Markéta
authors.initials: M
authors.lastname: Svobodová
authors.affiliation: Nanobiotechnology and Bioanalysis Group, Department of Chemical Engineering, Universitat Rovira I Virgili, 43007, Tarragona, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Mairal
authors.affiliation: 2bind GmbH, Josef Engert Strasse 13, 93053, Regensburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schubert
authors.affiliation: Life and Medical Sciences Institute, University of Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Künne
authors.affiliation: Life and Medical Sciences Institute, University of Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany.
authors.forename: Günter
authors.initials: G
authors.lastname: Mayer
authors.affiliation: Nanobiotechnology and Bioanalysis Group, Department of Chemical Engineering, Universitat Rovira I Virgili, 43007, Tarragona, Spain. ckosulli@etse.urv.es.|Institució Catalana de Recerca I Estudis Avancats, Passeig Lluís Companys 23, 08010, Barcelona, Spain. ckosulli@etse.urv.es.
authors.forename: Ciara K
authors.initials: CK
authors.lastname: O'Sullivan
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001665:Binding Sites; D007700:Kinetics; D031297:Lupinus / Q000737:chemistry*; D052156:SELEX Aptamer Technique / Q000379:methods; D055314:Seed Storage Proteins / Q000737:chemistry*
chemical_list: D052157:Aptamers, Nucleotide; D055314:Seed Storage Proteins
keywords: Dual aptamer; SELEX; Sandwich assay; Truncation studies; β-Conglutin