Analyte injection system
First Claim
1. A method of separating a first component of interest from at least one second component in a sample comprising:
- stacking the first and second components in a first channel segment;
flowing the stacked second component through a second channel segment fluidly coupled to the first channel segment at an intersection;
detecting a preselected electrical signal at or near the intersection which corresponds to the first and/or second stacked component; and
, applying an electric field or a pressure differential along a third channel segment which is fluidly coupled to said first channel segment at the intersection when the preselected electrical signal is detected, thereby introducing the stacked first component into the third channel segment.
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Abstract
This invention provides methods and devices for spatially separating at least first and second components in a sample which in one exemplary embodiment comprises introducing the first and second components into a first microfluidic channel of a microfluidic device in a carrier fluid comprising a spacer electrolyte solution and stacking the first and second components by isotachophoresis between a leading electrolyte solution and a trailing electrolyte solution, wherein the spacer electrolyte solution comprises ions which have an intermediate mobility in an electric field between the mobility of the ions present in the leading and trailing electrolyte solutions and wherein the spacer electrolyte solution comprises at least one of the following spacer ions MOPS, MES, Nonanoic acid, D-Glucuronic acid, Acetylsalicyclic acid, 4-Ethoxybenzoic acid, Glutaric acid, 3-Phenylpropionic acid, Phenoxyacetic acid, Cysteine, hippuric acid, p-hydroxyphenylacetic acid, isopropylmalonic acid, itaconic acid, citraconic acid, 3,5-dimethylbenzoic acid, 2,3-dimethylbenzoic acid, p-hydroxycinnamic acid, and 5-br-2,4-dihydroxybenzoic acid, and wherein the first component comprises a DNA-antibody conjugate and the second component comprises a complex of the DNA-antibody conjugate and an analyte.
148 Citations
58 Claims
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1. A method of separating a first component of interest from at least one second component in a sample comprising:
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stacking the first and second components in a first channel segment;
flowing the stacked second component through a second channel segment fluidly coupled to the first channel segment at an intersection;
detecting a preselected electrical signal at or near the intersection which corresponds to the first and/or second stacked component; and
,applying an electric field or a pressure differential along a third channel segment which is fluidly coupled to said first channel segment at the intersection when the preselected electrical signal is detected, thereby introducing the stacked first component into the third channel segment. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 23)
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22. A method of separating a first component of interest in a sample into separated components and detecting the separated components while minimizing interference during detecting from at least one second component in the sample, the method comprising introducing the sample into a separation channel and applying an electric field along a length of the separation channel to separate the first component of interest into separated components according to their electrophoretic mobilities while concomitantly stacking the second component in the separation channel between a leading electrolyte and a trailing electrolyte solution, and detecting the separated components.
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24. A microfluidic device comprising a main channel comprising an ITP stacking channel region and a separation channel region;
- and at least first and second side channels which are fluidly coupled to the main channel at a common fluid junction at the intersection of the ITP stacking channel region with the separation channel region, the first and second side channels terminating in first and second fluid reservoirs, respectively.
- View Dependent Claims (25, 26, 27, 28, 29)
- 30. A method of spatially separating at least first and second components in a sample in a microfluidic device comprising introducing the first and second components into a first microfluidic channel of the device in a carrier fluid comprising a spacer electrolyte solution and stacking the first and second components by isotachophoresis between a leading electrolyte solution and a trailing electrolyte solution, wherein the spacer electrolyte solution comprises ions which have an intermediate mobility in an electric field between the mobility of the ions present in the leading and trailing electrolyte solutions and wherein the spacer electrolyte solution comprises at least one of the following spacer ions MOPS, MES, Nonanoic acid, D-Glucuronic acid, Acetylsalicyclic acid, 4-Ethoxybenzoic acid, Glutaric acid, 3-Phenylpropionic acid, Phenoxyacetic acid, Cysteine, hippuric acid, p-hydroxyphenylacetic acid, isopropylmalonic acid, itaconic acid, citraconic acid, 3,5-dimethylbenzoic acid, 2,3-dimethylbenzoic acid, p-hydroxycinnamic acid, and 5-br-2,4-dihydroxybenzoic acid, and wherein the first component comprises a DNA-antibody conjugate and the second component comprises a complex of the DNA-antibody conjugate and an analyte.
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43. A method of separating a first component of interest from at least one second component in a sample comprising:
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stacking the first and second components in a first channel segment;
flowing the stacked second component through a second channel segment fluidly coupled to the first channel segment at an intersection;
measuring a voltage signal profile at or near the intersection which corresponds to the first and/or second stacked components, wherein the voltage signal profile includes at least three distinct voltage slope transitions which are separated in time; and
,applying an electric field or a pressure differential along a third channel segment which is fluidly coupled to said first channel segment at the intersection when the last in time voltage slope transition is detected, thereby introducing the stacked first component into the third channel segment. - View Dependent Claims (44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58)
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Specification