Dielectrophoretic separation and immunoassay methods on active electronic matrix devices
First Claim
1. A method for isolating and immobilizing at least one bioparticle of interest on an active electronic matrix chip device, wherein the device comprises:
- a substrate, individually addressable electrodes on the substrate, and a permeation layer overlying a plurality of the electrodes on the substrate, further wherein portions of the permeation layer over the electrodes form microlocations of the active electronic matrix chip device, further wherein at least one capture immunoreagent specific for the bioparticle of interest is attached to the permeation layer of the device at or between a plurality of microlocations, the method comprising;
a) introducing onto the active electronic matrix device a sample solution containing the bioparticle of interest, wherein the sample solution is of a conductivity suitable for dielectrophoretic isolation of the bioparticle of interest;
b) passing an alternating current through selected electrodes on the active electronic matrix chip device, wherein the electrodes are selected to produce areas of relatively high alternating current field strength and relatively low alternating current field strength at positions on the active electronic matrix chip device, wherein the alternating current is supplied at a suitable voltage and frequency for dielectrophoretic isolation of the bioparticle of interest, and further wherein the at least one capture immunoreagent specific for the bioparticle of interest are located at one or more positions of alternating current field strength at which the bioparticle of interest is predicted to aggregate; and
c) maintaining the alternating current in (b) for a sufficient length of time to allow the at least one capture immunoreagent to bind to the bioparticle of interest, thereby immobilizing the bioparticle.
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Abstract
This invention relates to devices and methods for performing active, multi-step molecular and biological sample preparation and diagnostic analyses employing immunochemical techniques. It relates generally to bioparticle separation, bioparticle enrichment, and electric field-mediated immunochemical detection on active electronic matrix devices utilizing AC and DC electric fields. More specifically, the invention relates to devices and methods for sample preparation/manipulation, immunoimmobilization, and immunoassays, all of which can be conducted on one or more active electronic chip devices within a single system. These manipulations are useful in a variety of applications, including, for example, detection of pathogenic bacteria and biological warfare agents, point-of-care diagnostics, food or medical product quality control assays, and other biological assays.
127 Citations
14 Claims
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1. A method for isolating and immobilizing at least one bioparticle of interest on an active electronic matrix chip device, wherein the device comprises:
- a substrate, individually addressable electrodes on the substrate, and a permeation layer overlying a plurality of the electrodes on the substrate, further wherein portions of the permeation layer over the electrodes form microlocations of the active electronic matrix chip device, further wherein at least one capture immunoreagent specific for the bioparticle of interest is attached to the permeation layer of the device at or between a plurality of microlocations, the method comprising;
a) introducing onto the active electronic matrix device a sample solution containing the bioparticle of interest, wherein the sample solution is of a conductivity suitable for dielectrophoretic isolation of the bioparticle of interest;
b) passing an alternating current through selected electrodes on the active electronic matrix chip device, wherein the electrodes are selected to produce areas of relatively high alternating current field strength and relatively low alternating current field strength at positions on the active electronic matrix chip device, wherein the alternating current is supplied at a suitable voltage and frequency for dielectrophoretic isolation of the bioparticle of interest, and further wherein the at least one capture immunoreagent specific for the bioparticle of interest are located at one or more positions of alternating current field strength at which the bioparticle of interest is predicted to aggregate; and
c) maintaining the alternating current in (b) for a sufficient length of time to allow the at least one capture immunoreagent to bind to the bioparticle of interest, thereby immobilizing the bioparticle. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
- a substrate, individually addressable electrodes on the substrate, and a permeation layer overlying a plurality of the electrodes on the substrate, further wherein portions of the permeation layer over the electrodes form microlocations of the active electronic matrix chip device, further wherein at least one capture immunoreagent specific for the bioparticle of interest is attached to the permeation layer of the device at or between a plurality of microlocations, the method comprising;
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9. A method for isolating and detectably labeling at least one bioparticle of interest on an active electronic matrix chip device, wherein the device comprises:
- a substrate, individually addressable electrodes on the substrate, and a permeation layer overlying a plurality of the electrodes on the substrate, further wherein portions of the permeation layer over the electrodes form microlocations of the active electronic matrix chip device, the method comprising;
a) introducing onto the active electronic matrix device a sample solution containing the bioparticle of interest, wherein the sample solution is of a conductivity suitable for dielectrophoretic isolation of the bioparticle of interest;
b) passing an alternating current through selected electrodes on the active electronic matrix chip device, wherein the electrodes are selected to produce areas of relatively high alternating current field strength and relatively low alternating current field strength at positions on the active electronic matrix chip device, wherein the alternating current is supplied at a suitable voltage and frequency for dielectrophoretic isolation of the bioparticle of interest, and further wherein one or more positions of alternating current field strength at which the bioparticle of interest is predicted to aggregate are at one or more aggregate microlocations of the active electronic matrix chip device;
c) maintaining the alternating current in (b) for a sufficient length of time to allow the bioparticle of interest to aggregate at the aggregate microlocations;
d) introducing onto the active electronic matrix chip device a solution comprising a detection immunoreagent specific for the bioparticle of interest;
e) passing a direct current through one or more aggregate microlocations, wherein the electrodes under the aggregate microlocations are biased so as to attract the detection immunoreagent to the aggregate microlocations from the solution; and
f) maintaining the direct current in (e) for a sufficient time to allow the detection immunoreagent to bind to the bioparticle of interest at the aggregate microlocation, thereby detectably labeling the bioparticle. - View Dependent Claims (10, 11, 12, 13, 14)
- a substrate, individually addressable electrodes on the substrate, and a permeation layer overlying a plurality of the electrodes on the substrate, further wherein portions of the permeation layer over the electrodes form microlocations of the active electronic matrix chip device, the method comprising;
Specification