Method for detecting multiple predetermined compounds in a sample
First Claim
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1. A method for detecting interaction of at least two predetermined compounds which comprises:
- (a) contacting at least a first predetermined compound present in a sample with a detectable tag moiety, wherein the detectable tag moiety comprises at least one detectable component, under conditions permitting the tag moiety to be conjugated to the predetermined compound present in the sample to form a detectably tagged predetermined compound;
(b) treating the sample from step (a) to form an electrolyte solution containing the detectably tagged predetermined compounds;
(c) contacting the electrolyte solution from step (b) with at least one nanopore, wherein at least a second predetermined compound is immobilized on the top of said at least one nanopore, under conditions permitting the binding of the detectably tagged predetermined compound with the second predetermined compound;
(d) treating the detectably tagged predetermined compound bound to said at least one nanopore so as to separate the tag moiety from the predetermined compound; and
(e) detecting said at least one detectable component of the tag moiety so released as the tag moiety passes through said at least one nanopore so as to thereby detect the interaction of the predetermined compounds.
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Abstract
This invention provides methods for detecting the presence of a plurality of predetermined compounds in a sample using a plurality of tag moieties and at least one nanopore. This invention also provides methods for determining the quantity of each of a plurality of predetermined compounds in a sample using a plurality of tag moieties and at least one nanopore. This invention further provides methods for detecting interaction of at least two predetermined compounds using at least one tag moiety and at least one nanopore.
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25 Claims
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1. A method for detecting interaction of at least two predetermined compounds which comprises:
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(a) contacting at least a first predetermined compound present in a sample with a detectable tag moiety, wherein the detectable tag moiety comprises at least one detectable component, under conditions permitting the tag moiety to be conjugated to the predetermined compound present in the sample to form a detectably tagged predetermined compound; (b) treating the sample from step (a) to form an electrolyte solution containing the detectably tagged predetermined compounds; (c) contacting the electrolyte solution from step (b) with at least one nanopore, wherein at least a second predetermined compound is immobilized on the top of said at least one nanopore, under conditions permitting the binding of the detectably tagged predetermined compound with the second predetermined compound; (d) treating the detectably tagged predetermined compound bound to said at least one nanopore so as to separate the tag moiety from the predetermined compound; and (e) detecting said at least one detectable component of the tag moiety so released as the tag moiety passes through said at least one nanopore so as to thereby detect the interaction of the predetermined compounds.
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2. A method for detecting interaction of at least two predetermined compounds which comprises:
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(a) contacting at least a first predetermined compound present in a sample with a detectable tag moiety, wherein the detectable tag moiety comprises at least one detectable component, under conditions permitting the tag moiety to be conjugated to the predetermined compound present in the sample to form a detectably tagged predetermined compound; (b) treating the sample from step (a) to form an electrolyte solution containing the detectably tagged predetermined compound; (c) contacting the electrolyte solution from step (b) with at least one nanopore, wherein at least a second predetermined compound is immobilized on the top of said at least one nanopore, under conditions permitting the binding of the detectably tagged predetermined compound with the second predetermined compound, and wherein the tag moiety of said detectably tagged predetermined compound bound to the second predetermined compound localizes within said at least one nanopore; (d) detecting said at least one detectable component of the tag moiety with said at least one nanopore so as to thereby detect the interaction of the predetermined compounds.
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3. A method for detecting the presence and quantity of a plurality of predetermined compounds in a plurality of samples, comprising:
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(a) contacting each sample with a nanopore-detectable tag moiety wherein the detectable tad moiety comprises at least one detectable component, under conditions permitting the tag moiety to be conjugated to the plurality of predetermined compounds present in the sample to form detectably tagged predetermined compounds for that sample, which are distinguishable from any nanopore-detectable tag moiety attached to similar predetermined compounds from any other sample; (b) mixing the resulting tagged compounds from the plurality of samples from step (a) and treating them in such a way as to form an electrolyte solution containing the set of detectably and distinguishably tagged predetermined compounds; (c) contacting the electrolyte solution from step (b) with at least one nanopore, wherein said at least one nanopore comprises a means on the top of said at least one nanopore for specific analyte moiety binding to each of the detectably tagged predetermined compounds in the electrolyte solution, under conditions such that the detectably tagged predetermined compounds bind to said at least one nanopore, thereby immobilizing the detectably tagged predetermined compounds on the top of the at least one nanopore; (d) treating the detectably tagged predetermined compounds bound to said at least one nanopore so as to separate the tag moieties from the predetermined compounds; and (e) detecting said at least one detectable component of each of the tag moieties so released as the tag moieties pass through said at least one nanopore so as to thereby detect the presence of the plurality of predetermined compounds in the plurality of samples (f) determining the amount of said at least one detectable component of each of the tag moieties so released as the tag moieties pass through said at least one nanopore; and (g) determining, from the amounts of said detectable components determined in step (e), the quantity of each of the plurality of predetermined compounds present in the sample. - View Dependent Claims (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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Specification