Magnetic beads-based electrochemical immunosensor for monitoring allergenic food proteins |
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Authors: | Michaela Čadková ,Radovan Metelka,Lucie Holubová ,Daniel Horá k,Veronika Dvořá ková ,Zuzana Bí lková ,Lucie Korecká |
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Affiliation: | 1. Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic;2. Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech Republic;3. Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, 162 06 Prague, Czech Republic |
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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 222 nM and a detection limit of 5 nM. 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. |
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Keywords: | Electrochemical immunosensor Magnetic particles Ovalbumin |
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