Process to detect binding events on an electrode microarray using enzymes
First Claim
1. A process to detect binding events on an electrode microarray using enzymes comprising:
- (a) providing a microarray having a plurality of electrodes and a plurality of capture complexes of at least two different types and attached at sites corresponding to the electrodes, wherein a first capture complex type has affinity for a first analyte type and subsequent capture complex types have affinity for subsequent analyte types, wherein the electrodes are electronically addressable;
(b) administering a plurality of analytes of at least one type to the capture complexes to form bound complexes of types corresponding to each type of the analytes captured by the capture complexes;
(c) attaching enzymes to the bound complexes to form reporter complexes, wherein the reporter complexes are of a type corresponding to each type of the analytes captured by the capture complexes;
(d) sequentially contacting a plurality of substrate solutions to the microarray and measuring for the presence or absence of an enzymatic product at the sites using an electrical signal, wherein each substrate solution corresponds to the reporter complex type, wherein the presence of the electrical signal is a measure of the binding event.
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Abstract
The present invention provides a process to detect binding events on an electrode microarray. A microarray is provided having addressable electrodes and two or more different types of capture complexes at sites corresponding to the electrodes. The capture complexes capture analytes. Enzymes are attached to form a reporter complex. Substrate solutions are sequentially contacted to make enzyme products that are detectable at the electrodes by a difference in the electrical response at electrodes having the enzyme product and those not having the enzyme product. The enzyme product may be a solid deposition product. The electrical properties of electrodes on the microarray are read for the presence of the enzyme product by sequentially switching each electrode held at a constant voltage to ground and then back to the constant voltage.
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Citations
33 Claims
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1. A process to detect binding events on an electrode microarray using enzymes comprising:
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(a) providing a microarray having a plurality of electrodes and a plurality of capture complexes of at least two different types and attached at sites corresponding to the electrodes, wherein a first capture complex type has affinity for a first analyte type and subsequent capture complex types have affinity for subsequent analyte types, wherein the electrodes are electronically addressable;
(b) administering a plurality of analytes of at least one type to the capture complexes to form bound complexes of types corresponding to each type of the analytes captured by the capture complexes;
(c) attaching enzymes to the bound complexes to form reporter complexes, wherein the reporter complexes are of a type corresponding to each type of the analytes captured by the capture complexes;
(d) sequentially contacting a plurality of substrate solutions to the microarray and measuring for the presence or absence of an enzymatic product at the sites using an electrical signal, wherein each substrate solution corresponds to the reporter complex type, wherein the presence of the electrical signal is a measure of the binding event. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A process to detect binding events on an electrode microarray using enzymes comprising:
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(a) providing a microarray having a plurality of electrodes and a plurality of capture complexes attached at sites corresponding to the electrodes, wherein the electrodes are electronically addressable;
(b) administering a plurality of analytes to the capture complexes to form bound complexes, wherein the capture complexes have affinity to the analytes;
(c) attaching enzymes to the bound complexes to form reporter complexes;
(d) contacting to the microarray a substrate solution having substrates reactive to the enzymes, wherein a solid enzyme product deposits on the sites having enzymes; and
(e) measuring for the presence or absence of the solid enzyme product at the sites using a measurement solution and an electrical signal, wherein the presence of the electrical signal is a measure of the binding event. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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31. A process to detect binding events on an electrode microarray using enzymes comprising:
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(a) providing a microarray having a plurality of electrodes, wherein the electrodes are electronically addressable;
(b) applying a constant initial voltage to the electrodes while maintaining an open circuit to the electrodes; and
(c) measuring an electrical response of each electrode by sequentially switching each electrode to approximately ground or a voltage substantially different from the constant initial voltage for a measurement time, recording current flow at each electrode during the measurement time, and returning each electrode to the constant initial voltage before setting the next electrode to ground or the voltage substantially different from the constant initial voltage. - View Dependent Claims (32, 33)
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Specification