METHODS FOR GENERATING pH/IONIC CONCENTRATION GRADIENT NEAR ELECTRODE SURFACES FOR MODULATING BIOMOLECULAR INTERACTIONS
First Claim
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1. A method of modulating the pH or ionic concentration in a biosensor, the method comprising:
- a) providing a biosensor comprising a multisite array of test sites in which the conditions for interacting with a biomolecule analyte can be varied independently, each test site comprising a support in an aqueous solution, the support comprising one or more electrodes or an electromagnet, and a biomolecule interface layer having one or more immobilized probes thereon;
b) adding an electrochemically active agent, an enzyme, an enzyme substrate, a buffer inhibitor, or a combination thereof to the aqueous solution; and
c) reacting the electrochemically active agent, the enzyme, the enzyme substrate, or combination thereof in the aqueous solution to produce H+ ion or OH−
ions, or increasing the diffusion of H+ ions or OH−
ions with the buffer inhibitor, or inhibiting the interaction between H+ ions or OH−
ions and buffering salts with the buffer inhibitor.
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Abstract
Methods of modulating the binding interactions between a (biomolecular) probe or detection agent and an analyte of interest from a biological sample in a biosensor having a multisite array of test sites. In particular, the methods modulate the pH or ionic concentration gradient near the electrodes in such biosensor. The methods of modulating the binding interactions provide a biosensor and analytic methods for more accurately measuring an analyte of interest in a biological sample.
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Citations
34 Claims
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1. A method of modulating the pH or ionic concentration in a biosensor, the method comprising:
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a) providing a biosensor comprising a multisite array of test sites in which the conditions for interacting with a biomolecule analyte can be varied independently, each test site comprising a support in an aqueous solution, the support comprising one or more electrodes or an electromagnet, and a biomolecule interface layer having one or more immobilized probes thereon; b) adding an electrochemically active agent, an enzyme, an enzyme substrate, a buffer inhibitor, or a combination thereof to the aqueous solution; and c) reacting the electrochemically active agent, the enzyme, the enzyme substrate, or combination thereof in the aqueous solution to produce H+ ion or OH−
ions, or increasing the diffusion of H+ ions or OH−
ions with the buffer inhibitor, or inhibiting the interaction between H+ ions or OH−
ions and buffering salts with the buffer inhibitor. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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26. A biosensor for use in detecting a biomolecule analyte comprising a multisite array of test sites in which the conditions for interacting with the biomolecule analyte can be varied independently, each test site comprising:
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a) a support in an aqueous environment; b) one or more electrodes; and c) a biomolecule interface layer having one or more immobilized detection agents and one or more immobilized enzymes. - View Dependent Claims (27, 28, 29, 30, 31)
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32. A method for detecting a biomolecule analyte in a biological sample, the method comprising:
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a) providing a biosensor comprising a multisite array of test sites in which the conditions for interacting with a biomolecule analyte can be varied independently, each test site comprising a support in an aqueous solution, the support comprising one or more electrodes or an electromagnet, and a biomolecule interface layer having one or more immobilized probes thereon; b) adding in each test site an electrochemically active agent, an enzyme, an enzyme substrate, a buffer inhibitor, or a combination thereof to the aqueous solution; c) reacting the electrochemically active agent, the enzyme, the enzyme substrate, or a combination thereof in the aqueous solution to produce H+ ion or OH−
ions, or increasing the diffusion of H+ ions or OH−
ions with a buffer inhibitor, or inhibiting the interaction between H+ ions or OH−
ions and buffering salts with the buffer inhibitor;d) adding a biological sample to each test site; and e) detecting the biomolecule analyte in each test site, wherein the amounts added in step b) and the reaction in step c) are varied between test sites in a subset array of test sites in order to obtain sets of test sites in which the pH or ionic concentration near electrode surfaces in the test sites varies. - View Dependent Claims (33, 34)
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Specification