Process for specific binding assay for measuring the amount of analyte in a liquid test sample
First Claim
1. A specific binding assay process for measuring the amount of a substance to be assayed in a liquid test sample, comprising the steps of:
- (a) providing a device comprising;
a matrix in which a distribution of a labeled specific binding substance forms, a sample introduction site connected to said matrix, a detection site in contact with substantially the most upstream side or most downstream side of said matrix for detecting a signal which is generated when a signal substance contacts said detection site, and an absorption site in which said liquid test sample flows from said sample introduction site through said matrix;
(b) introducing said labeled specific binding substance and said liquid test sample into said matrix through said sample introduction site simultaneously or sequentially, wherein said labeled specific binding substance comprises a specific binding substance and a label, and wherein said labeled specific binding substance specifically competes with said substance to be assayed for a second specific binding substance and generates said signal substance in the presence of at least one substance related to generation of a signal substance, wherein said second specific binding substance preliminarily exists in the matrix or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, causing said labeled specific binding substance to flow with the liquid test sample through said matrix, and causing said labeled specific binding substance to compete with said substance to be assayed in the matrix for said second specific binding substance, wherein binding to the second specific binding substance causes a difference in the mobility of said labeled specific binding substance in a bound state from the mobility of said labeled specific binding substance in an unbound state, thereby forming a distribution of bound and unbound labeled specific binding substance, wherein the distribution depends upon the amount of said substance to be assayed in said liquid test sample, causing said label of said labeled specific binding substance to directly or indirectly produce said signal substance with the reaction with at least one substance related to generation of a signal substance, wherein said substance related to generation of a signal substance preliminarily exists in the matrix or the absorption site, or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, and allowing said signal substance to diffuse independently of the flow direction of said labeled specific binding substance to reach said detection site;
(c) measuring the intensity of said signal generated from said signal substance which reaches the detection site, wherein said intensity reflects the distribution of said labeled specific binding substance upon signal generation; and
(d) determining the amount of said substance to be assayed by comparing said intensity to reference values;
wherein said signal is an electrochemical signal and wherein said labeled specific binding substance is not immobilized in the matrix.
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Abstract
A specific binding assay process which is useful for quick qualitative or quantitative measurement and which can be used for various purposes, and a specific binding assay device suitable for the practice of the process,
in which a substance to be assayed in a liquid test sample is determined qualitatively or quantitatively by developing the substance in a liquid test sample in a matrix, which comprises the steps of, for example:
allowing the substance to be assayed to reach with a specific binding substance which has specific affinity for the substance to be assayed;
allowing a signal substance generator (a substance which competes with the substance to be assayed for the specific binding substance and which generates a signal) to change its distribution in the matrix in response to the reaction of the above step, and;
detecting the resulting distributional changes at a detection means as changes of signals which are rate-limited by the mass transfer of a signal substance generated from the signal substance generator.
87 Citations
4 Claims
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1. A specific binding assay process for measuring the amount of a substance to be assayed in a liquid test sample, comprising the steps of:
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(a) providing a device comprising;
a matrix in which a distribution of a labeled specific binding substance forms, a sample introduction site connected to said matrix, a detection site in contact with substantially the most upstream side or most downstream side of said matrix for detecting a signal which is generated when a signal substance contacts said detection site, and an absorption site in which said liquid test sample flows from said sample introduction site through said matrix;
(b) introducing said labeled specific binding substance and said liquid test sample into said matrix through said sample introduction site simultaneously or sequentially, wherein said labeled specific binding substance comprises a specific binding substance and a label, and wherein said labeled specific binding substance specifically competes with said substance to be assayed for a second specific binding substance and generates said signal substance in the presence of at least one substance related to generation of a signal substance, wherein said second specific binding substance preliminarily exists in the matrix or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, causing said labeled specific binding substance to flow with the liquid test sample through said matrix, and causing said labeled specific binding substance to compete with said substance to be assayed in the matrix for said second specific binding substance, wherein binding to the second specific binding substance causes a difference in the mobility of said labeled specific binding substance in a bound state from the mobility of said labeled specific binding substance in an unbound state, thereby forming a distribution of bound and unbound labeled specific binding substance, wherein the distribution depends upon the amount of said substance to be assayed in said liquid test sample, causing said label of said labeled specific binding substance to directly or indirectly produce said signal substance with the reaction with at least one substance related to generation of a signal substance, wherein said substance related to generation of a signal substance preliminarily exists in the matrix or the absorption site, or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, and allowing said signal substance to diffuse independently of the flow direction of said labeled specific binding substance to reach said detection site;
(c) measuring the intensity of said signal generated from said signal substance which reaches the detection site, wherein said intensity reflects the distribution of said labeled specific binding substance upon signal generation; and
(d) determining the amount of said substance to be assayed by comparing said intensity to reference values;
wherein said signal is an electrochemical signal and wherein said labeled specific binding substance is not immobilized in the matrix.
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2. A specific binding assay process for measuring the amount of a substance to be assayed in a liquid test sample, comprising the steps of:
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(a) providing a device comprising;
a matrix in which a distribution of a labeled specific binding substance forms, a sample introduction site connected to said matrix, and a detection site in contact with substantially the most upstream side or most downstream side of said matrix for detecting a signal, which is generated when a signal substance contacts said detection site;
(b) introducing said labeled specific binding substance and said liquid test sample into said matrix through said sample introduction site simultaneously or sequentially, wherein said labeled specific binding substance comprises a specific binding substance and a label, and wherein said labeled specific binding substance specifically competes with said substance to be assayed for a second specific binding substance and generates said signal substance in the presence of at least one substance related to generation of a signal substance, wherein said second specific binding substance preliminarily exists in the matrix or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, causing said labeled specific binding substance to flow with the liquid test sample through said matrix, and causing said labeled specific binding substance to compete with said substance to be assayed in the matrix for said second specific binding substance, wherein binding to the second specific binding substance causes a difference in the mobility of said labeled specific binding substance in a bound state from the mobility of said labeled specific binding substance in an unbound state, thereby forming a distribution of bound and unbound labeled specific binding substance, wherein the distribution depends upon the amount of said substance to be assayed in said liquid test sample, causing said label of said labeled specific binding substance to directly or indirectly produce said signal substance, with the reaction with at least one substance related to generation of a signal substance, wherein said substance related to generation of a signal substance preliminarily exists in the matrix or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, and allowing said signal substance to diffuse independently of the flow direction of said labeled specific binding substance to reach said detection site;
(c) measuring the intensity of said signal generated from said signal substance which reaches the detection site, wherein said intensity reflects the distribution of said labeled specific binding substance upon signal generation; and
(d) determining the amount of said substance to be assayed by comparing said intensity to reference values;
wherein said signal is an electrochemical signal and wherein said labeled specific binding substance is not immobilized in the matrix.
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3. A specific binding assay process for measuring the amount of a substance to be assayed in a liquid test sample, comprising the steps of:
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(a) providing a device comprising;
a matrix in which a distribution of a labeled specific binding substance forms, a sample introduction site connected to said matrix, and a detection site in contact with substantially the most upstream side or most downstream side of said matrix for detecting a signal which is generated when a signal substance contacts said detection site;
(b) introducing said labeled specific binding substance and said liquid test sample into said matrix through said sample introduction site simultaneously or sequentially, wherein said labeled specific binding substance comprises a specific binding substance and a label, and wherein said labeled specific binding substance directly or indirectly binds to said substance to be assayed and generates said signal substance in the presence of at least one substance related to generation of a signal substance, causing said labeled specific binding substance to flow with the liquid test sample through said matrix, and causing said labeled specific binding substance to bind to said substance to be assayed in the matrix, wherein the mobility of said labeled specific binding substance in a bound state differs from the mobility of said labeled specific binding substance in an unbound state, thereby forming a distribution of bound and unbound labeled specific binding substance, wherein the distribution depends upon the amount of said substance to be assayed in said liquid test sample, causing said label of said labeled specific binding substance to directly or indirectly produce said signal substance with the reaction with at least one substance related to generation of a signal substance, wherein said substance related to generation of a signal substance, wherein said substance related to the generation of a signal substance preliminarily exists in the matrix or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, and allowing said signal substance to diffuse independently of the flow direction of said labeled specific binding substance to reach said detection site;
(c) measuring the intensity of said signal generated from said signal substance which reaches the detection site, wherein said intensity reflects the distribution of said labeled specific binding substance upon signal generation; and
(d) determining the amount of said substance to be assayed by comparing said intensity to reference values;
wherein said signal is an electrochemical signal and wherein said labeled specific binding substance is not immobilized in the matrix.
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4. A specific binding assay process for measuring the amount of a substance to be assayed in a liquid test sample, comprising the steps of:
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(a) providing a device comprising;
a matrix in which a distribution of a labeled specific binding substance forms, a sample introduction site connected to said matrix, a detection site in contact with substantially the most upstream side or the most downstream side of said matrix for detecting a signal which is generated when a signal substance contacts said detection site, and an absorption site in which said liquid test sample flows from said sample introduction site through said matrix;
(b) introducing said labeled specific binding substance and said liquid test sample into said matrix through said sample introduction site simultaneously or sequentially, wherein said labeled specific binding substance comprises a specific binding substance and a label, and wherein said labeled specific binding substance directly or indirectly binds to said substance to be assayed and generates said signal substance in the presence of at least one substance related to generation of a signal substance wherein said substance related to generation of a signal substance preliminarily exists in the matrix or absorption site, or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, causing said labeled specific binding substance to flow with the liquid test sample through said matrix, and causing said labeled specific binding substance to directly or indirectly bind to said substance to be assayed in the matrix, wherein the mobility of said labeled specific binding substance in a bound state differs from the mobility of said labeled specific binding substance in an unbound state, thereby forming a distribution of bound and unbound labeled specific binding substance, wherein the distribution depends upon the amount of said substance to be assayed in said liquid test sample, causing said label of said labeled specific binding substance to directly or indirectly produce said signal substance with the reaction with at least one substance related to generation of a signal substance, wherein said substance related to generation of a signal substance preliminarily exists in the matrix or is introduced into the matrix through said sample introduction site with the simultaneous or sequential addition of the test sample, and allowing said signal substance to diffuse independently of the flow direction of said labeled specific binding substance to reach said detection site;
(c) measuring the intensity of said signal generated from said signal substance which reaches the detection site, wherein said intensity reflects the distribution of said labeled specific binding substance upon signal generation; and
(d) determining the amount of said substance to be assayed by comparing said intensity to reference values;
wherein said Anal is an electrochemical signal and wherein said labeled specific binding substance is not immobilized in the matrix.
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Specification