title: Magnetic beads-based electrochemical immunosensor for monitoring allergenic food proteins.
abstract: Screen-printed platinum electrodes as transducer and magnetic beads as solid phase were combined to develop a particle-based electrochemical immunosensor for monitoring the serious food allergen ovalbumin. The standard arrangement of enzyme-linked immunosorbent assay became the basis for designing the immunosensor. A sandwich-type immunocomplex was formed between magnetic particles functionalized with specific anti-ovalbumin immunoglobulin G and captured ovalbumin molecules, and secondary anti-ovalbumin antibodies conjugated with the enzyme horseradish peroxidase were subsequently added as label tag. The electrochemical signal proportional to the enzymatic reaction of horseradish peroxidase during the reduction of hydrogen peroxide with thionine as electron mediator was measured by linear sweep voltammetry. The newly established method of ovalbumin detection exhibits high sensitivity suitable for quantification in the range of 11 to 222nM and a detection limit of 5nM. Magnetic beads-based assay format using external magnets for rapid and simple separation has been proven to be an excellent basis for electrochemical detection and quantification of food allergens in highly complex sample matrices.
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Čadková
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic.
authors.forename: Radovan
authors.initials: R
authors.lastname: Metelka
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Holubová
authors.affiliation: Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, 162 06 Prague, Czech Republic.
authors.forename: Daniel
authors.initials: D
authors.lastname: Horák
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic.
authors.forename: Veronika
authors.initials: V
authors.lastname: Dvořáková
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Bílková
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic. Electronic address: lucie.korecka@upce.cz.
authors.forename: Lucie
authors.initials: L
authors.lastname: Korecká
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004553:Electric Conductivity; D004563:Electrochemistry; D004566:Electrodes; D004579:Electron Transport; D005512:Food Hypersensitivity / Q000378:metabolism*; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D057230:Limit of Detection; D059346:Magnets / Q000737:chemistry*; D008863:Microspheres; D010047:Ovalbumin / Q000009:adverse effects / Q000032:analysis*; D010984:Platinum / Q000737:chemistry
chemical_list: D055910:Antibodies, Immobilized; D010984:Platinum; D010047:Ovalbumin
keywords: Electrochemical immunosensor; Magnetic particles; Ovalbumin