Biosensor with improved interference characteristics
First Claim
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1. A device for detecting a target analyte, the device comprising:
- an electrochemical cell;
an electron redox mediating molecule; and
a recombinant modified enzyme engineered to;
prevent transfer of electrons to a natural electron acceptor;
transfer electrons to the electron redox mediating molecule; and
provide enzymatic activity against the target analyte to generate an electrochemical signal dependent on a concentration of the target analyte in a sample.
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Abstract
This invention relates to mutagenized redox oxidase enzymes used to design enzyme electrodes with improved interference characteristics in the presence of mediator and oxygen in the assay. This recombinant modified enzyme has enhanced capability to transfer electrons to redox mediator instead of its natural electron acceptors such as O2, NAD, NADP, etc., which will improve the assay performance with less interference and higher sensitivity.
14 Citations
20 Claims
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1. A device for detecting a target analyte, the device comprising:
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an electrochemical cell; an electron redox mediating molecule; and a recombinant modified enzyme engineered to; prevent transfer of electrons to a natural electron acceptor; transfer electrons to the electron redox mediating molecule; and provide enzymatic activity against the target analyte to generate an electrochemical signal dependent on a concentration of the target analyte in a sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A device for detecting a target analyte, the device comprising:
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an electron redox mediating molecule; and a recombinant modified enzyme comprising a mutation of a protein coding sequence for a starting enzyme, wherein the mutation is engineered to modify an active site of the starting enzyme for oxygen reduction and electron transfer to a natural electron acceptor of the starting enzyme permitting transfer of electrons to the electron redox mediating molecule, and the recombinant modified enzyme maintains enzymatic activity against the target analyte.
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20. An electrochemical biosensor, comprising:
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a silver/silver chloride electrode connected to one end of a circuit; a conductive metal surface connected to another end of a circuit; and an immobilized enzyme layer comprising; an electron mediator molecule; and a recombinant modified enzyme lacking an ability to transfer electrons to a natural electron acceptor and instead transfers electrons to the electron mediator molecule such that the biosensor performs in a presence of oxygen.
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Specification