Particle based electrochemiluminescent assays

A method for the detection and quantitation of an analyte of interest in a sample, which method comprises the steps of:

• (1) forming a composition comprising:
• (a) a sample, which may comprise a first quantity of the analyte of interest;
• (b) a known amount of at least one reagent substance selected from the group consisting of:
• (i) a second quantity of the analyte of interest or an analog of the analyte of interest, and
• (ii) a binding partner of the analyte of interest or its said analog,
• wherein one of said reagent substances is linked to a label compound capable of being induced to electrochemiluminesce, and
• (c) a known amount of a second reagent comprising particles which are:
• (i) capable of specifically binding with the analyte of interest or (ii) attached to the second quantity of the analyte of interest or its analog, wherein said particles and said analyte are different;
• (2) inducing the label compound to electrochemiluminesce;
• and
• (3) comparing the luminescence emitted by the label compound to the luminescence of a calibration standard to determine whether or not, or in what amount, the analyte of interest is present in the sample.

An assay method for determining the presence or amount of an analyte of interest based upon a specific binding reaction and the measurement of an electrochemiluminescent phenomenon comprising the steps of:

• (a) forming an assay mixture containing:
• (i) a sample containing a first quantity of the analyte of interest,
• (ii) at least one reagent substance selected from the group consisting of
• (A) a second quantity of the analyte of interest or an analog of the analyte of interest, and
• (B) a binding partner of the analyte of interest or its said analog,
• wherein one of said reagent substances is linked to a label compound having a chemical moiety capable of being induced to electrochemiluminesce, and
• (iii) a second reagent comprising a plurality of inanimate particles which are:
• (1) capable of specifically binding with the analyte of interest and/or (2) attached to the second quantity of the analyte or its analog;
• and
• (b) incubating said assay mixture to permit binding;
• (c) causing the label compound to electrochemiluminesce;
• and
• (d) measuring electrochemiluminescence emitted by said label compound and correlating said measurement to a calibration standard to determine whether or not, or in what amount the analyte of interest is present in the sample.

A homogeneous assay method for determining the presence or amount of an analyte of interest based upon a specific binding reaction and the measurement of an electrochemiluminescent phenomenon comprising the steps of:

• (a) forming an assay mixture containing:
• (i) a sample containing a first quantity of the analyte of interest,
• (ii) at least one reagent substance selected from the group consisting of:
• (A) a second quantity of the analyte of interest or an analog of the analyte of interest, and
• (B) a binding partner of the analyte of interest or its said analog,
• wherein one of said reagent substances is linked to a label compound having a chemical moiety capable of being induced to electrochemiluminesce;
• and
• (iii) a second reagent comprising a plurality of particles which are:
• (A) capable of specifically binding with the analyte and/or (B) attached to the second quantity of the analyte of interest or its analog, wherein said particles and said analyte are different;
• and
• (b) introducing said assay mixture into an electrochemiluminescence cell;
• (c) causing the label compound to electrochemiluminesce in said electrochemiluminescence cell;
• and
• (d) measuring electrochemiluminescence emitted by said label compound in the presence of said particles in said electrochemiluminescence cell and correlating said measurement to a calibration standard to determine whether or not, or in what amount, the analyte of interest is present in the sample.

An assay method for determining the presence or amount of an analyte of interest based upon a specific binding reaction and the measurement of an electrochemiluminescent phenomenon comprising the steps of:

• (a) forming an assay mixture containing:
• (i) a sample containing a first quantity of the analyte of interest,
• (ii) at least one reagent substance selected from the group consisting of:
• (A) a second quantity of the analyte of interest or an analog of the analyte of interest, and
• (B) a binding partner of the analyte of interest or its said analog,
• wherein one of said reagent substances is linked to a label compound having a chemical moiety capable of being induced to electrochemiluminesce;
• and
• (iii) a second reagent comprising a plurality of inanimate particles which are:
• (A) capable of specifically binding with the analyte or (B) attached to the second quantity of the analyte of interest or its analog;
• (b) introducing said assay mixture into an electrochemiluminescence cell;
• (c) causing the label compound to electrochemiluminesce in said electrochemiluminescence cell;
• and
• (d) measuring electrochemiluminescence emitted by said label compound in said electrochemiluminescence cell and correlating said measurement to a calibration standard to determine whether or not, or in what amount, the analyte of interest is present in the sample.

An assay composition for use in detecting an analyte of interest in a sample, comprising:

• (a) a sample containing the analyte of interest,
• (b) at least one reagent substance selected from the group consisting of
• (i) added analyte of interest or an analog of the analyte of interest,
• (ii) a binding partner of the analyte of interest or its said analog, and
• (iii) a reactive component capable of binding with (i) or (ii), wherein one of said reagent substances is linked to a label compound having a chemical moiety capable of being induced to electrochemiluminesce,
• (c) a second reagent comprising a plurality of inanimate particles capable of specifically binding with the analyte and/or at least one reagent substance defined in (b)(i), (b)(ii), or (b)(iii), and
• (d) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence.

A composition of matter for use as a reagent in a microparticulate-based electrochemiluminescence binding assay for determining the presence or amount of an analyte of interest, comprising (i) a reagent comprising microparticles which are capable of binding with said analyte of interest or linked to said analyte of interest or its analog, (ii) a label containing an ECL moiety, (iii) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence and (iv) at least one other assay component selected from the group consisting of:

• (a) electrolyte,
• (b) analyte of interest or an analog of the analyte of interest,
• (c) a binding partner of the analyte of interest or its analog,
• (d) a reactive component capable of reacting with (b) or (c), and
• (e) an electrochemiluminescent-reaction enhancer,
• provided, however, that no two components contained within said composition are reactive with one another during storage so as to impair their function in the intended assay and said composition is suitable for use in said microparticulate-based electrochemiluminescence binding assay.

A kit containing in one or more vessels reagents for use in a microparticulate-based electrochemiluminescence binding assay for determining the presence or amount of an analyte of interest comprising:

• (1) a label compound reagent containing an ECL moiety,
• (2) a reagent comprising microparticles which are:
• (i) capable of binding with said analyte of interest or a reactive component capable of binding with said analyte of interest or (ii) linked to said analyte of interest or its analog, and
• (3) at least one other assay reagent component selected from the group consisting of:
• (a) electrolyte,
• (b) analyte of interest or an analog of the analyte of interest,
• (c) a binding partner of the analyte of interest or its analog,
• (d) a reactive component capable of reacting with (b) or (c),
• (e) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence, and
• (f) an electrochemiluminescent reaction enhancer;
• provided, however, that no two of said reagents react with one another under storage conditions so as to impair the function of the reagents in the intended assay, and wherein said kit is suitable for use in said microparticle-based electrochemiluminescence binding assay.

A system for detecting or quantitating an analyte of interest in a sample based upon an electrochemiluminescent phenomenon comprising:

• (a) a sample;
• (b) at least one reagent substance selected from the group consisting of
• (i) added analyte of interest or an analog of the analyte of interest,
• (ii) a binding partner of the analyte of interest or its said analog, and
• (iii) a reactive component capable of binding with (i) or (ii), wherein one of said reagent substances is linked to a label compound having a chemical moiety capable of being induced to electrochemiluminesce;
• (c) a reagent comprising (i) a plurality of particles capable of specifically binding to the analyte or (ii) a plurality of particles linked to said analyte or its analog;
• (d) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence;
• (e) a voltage source for inducing the label compound to electrochemiluminesce;
• and
• (f) a light detector for measuring luminescence emitted by said system to determine the presence or quantity of the analyte of interest in the sample.

A heterogeneous assay method based upon a specific binding reaction and an electrochemiluminescent phenomenon comprising the steps of:

• (a) forming an assay mixture containing:
• (i) a sample containing a first quantity of an analyte of interest having binding properties;
• (ii) a reagent comprising a plurality of inanimate particles which (A) have a surface capable of specifically binding to said analyte of interest or (B) are attached to a second quantity of the analyte of interest or an analog of the analyte of interest;
• and
• (iii) a label reagent substance selected from the group consisting of (A) the second quantity of the analyte of interest or said analog of the analyte of interest, and (B) a binding partner of the analyte of interest or its said analog, said label reagent substance including a chemical moiety having electrochemiluminescent properties;
• (b) incubating said assay mixture to permit binding;
• (c) separating said particles from said assay mixture;
• (d) causing the label reagent substance to electrochemiluminesce in a electrochemiluminescence cell;
• and
• (e) measuring luminescence emitted by said label reagent substance and correlating said measurement to a standard to determine whether or not, or in what amount, the analyte of interest is present in the sample.

An assay composition for an assay based upon a binding reaction and an electrochemiluminescent phenomenon comprising:

• (a) an electrolyte,
• (b) a sample containing an analyte of interest having binding properties,
• (c) a reagent comprising a plurality of particles having a surface which is:
• (i) capable of binding to a component of said assay composition or (ii) is linked to said analyte of interest or its analog, wherein said particles and said analyte are different,
• (d) a label substance having binding properties, said label substance including a chemical moiety having electrochemiluminescent properties, and
• (e) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminesce,
• wherein said assay composition is suitable for use in said assay based on said electrochemiluminescent phenomenon.

An assay reagent for use in an assay method for detecting an analyte of interest in a sample using an electrochemiluminescent phenomenon, which assay reagent comprises:

• (a) at least one assay reagent component selected from the group consisting of:
• (i) analyte of interest or an analog of the analyte of interest,
• (ii) a binding partner of the analyte of interest or its said analog, and
• (iii) a reactive component capable of binding with (i) or (ii), wherein one of said assay reagent components is linked to a label compound having a chemical moiety capable of being induced to electrochemiluminesce,
• (b) a reagent comprising a plurality of suspended inanimate particles which are:
• (i) capable of specifically binding with the analyte or (ii) attached to the analyte or its analog, and
• (c) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence,
• provided that the components of said assay reagent are not reactive with one another during storage so as to impair their function in the intended assay and said assay reagent is suitable for use in said assay method using said electrochemiluminescent phenomenon.

An assay reagent for an assay based upon a binding reaction and an electrochemiluminescent phenomenon comprising:

• (a) an electrolyte,
• (b) a reagent comprising a plurality of particles which:
• (i) have a surface capable of binding to said analyte or (ii) are attached to said analyte or its analog,
• (c) a label substance selected from the group consisting of (A) added analyte of interest or its analog, (B) a binding partner of the analyte of interest or its said analog, and (C) a reactive component capable of binding with (A) or (B), said label substance including a chemical moiety having electrochemiluminescent properties, and
• (d) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence,
• wherein said assay reagent is suitable for use in said assay based on said electrochemiluminescent phenomenon.

A method for the detection or quantitation of an analyte of interest in a sample, which method comprises the steps of:

• (1) forming a composition comprising:
• (a) a sample, which may comprise a first quantity of the analyte of interest;
• (b) at least one reagent substance selected from the group consisting of:
• (i) a second quantity of the analyte of interest or an analog of the analyte of interest, and
• (ii) a binding partner of the analyte of interest or its said analog
• wherein one of said reagent substances is linked to a label compound capable of being induced to electrochemiluminesce;
• (c) a second reagent comprising a plurality of particles which are:
• (i) capable of specifically binding with the analyte of interest or (ii) attached to the second quantity of the analyte of interest or its analog, wherein said particles are selected from the group consisting of polymeric particles, inorganic particles, metallic particles, metal oxide particles, composite particles and mixtures thereof;
• (2) introducing said composition into an electrochemiluminescence cell;
• and
• (3) inducing the label compound to electrochemiluminesce in said composition in the presence of said particles in said electrochemiluminescence cell;
• and
• (4) measuring luminescence emitted by the label compound in the presence of said particles to determine whether or not, or in what amount, the analyte of interest is present in the sample.

A method for the detection and quantitation of an analyte of interest in a sample, which method comprises the steps of:

• (1) forming a composition comprising:
• (a) a sample,
• (b) a known amount of at least one reagent substance selected from the group consisting of:
• (i) added analyte of interest or an analog of the analyte of interest, and
• (ii) binding partner of the analyte of interest or its said analog, wherein one of said reagent substances is linked to a label compound capable of being induced to electrochemiluminesce, and
• (c) a known amount of a second reagent comprising particles which are:
• (i) capable of specifically binding with the analyte or (ii) attached to the analyte or its analog, wherein said particles and said analyte are different;
• (2) introducing said composition into an electrochemiluminescence cell;
• (3) inducing the label compound to electrochemiluminesce in said composition in the presence of said particles in said electrochemiluminescence cell;
• and
• (4) comparing the luminescence emitted by the label compound to the luminescence of a calibration standard to determine whether or not, or in what amount, the analyte of interest is present in the sample.

An assay reagent for an assay based upon a binding reaction and an electrochemiluminescent phenomenon comprising:

• (a) at least one reagent substance selected from the group consisting of:
• (i) added analyte of interest or an analog of the analyte of interest,
• (ii) a binding partner of the analyte of interest or its said analog, and
• (iii) a reactive component capable of binding with (i) or (ii) wherein one of said reagent substances is linked to a label compound capable of being induced to electrochemiluminesce,
• (c) a second reagent comprising a plurality of particles which (i) have a surface capable of binding with the analyte or (ii) are attached to the analyte or its analog, and
• (d) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminesce,
• wherein said assay reagent is suitable for use in said assay based on said electrochemiluminescent phenomenon.

An assay composition for use in an assay based upon a binding reaction and an electrochemiluminescent phenomenon comprising:

• (a) a sample;
• (b) at least one reagent substance selected from the group consisting of:
• (i) added analyte of interest or an analog of the analyte of interest,
• (ii) a binding partner of the analyte of interest or its said analog, and
• (iii) a reactive component capable of binding with (i) or (ii), wherein one of said reagent substances is linked to a label compound capable of being induced to electrochemiluminesce,
• (c) a second reagent comprising a plurality of particles which (i) have a surface capable of binding with the analyte or (ii) are attached to the analyte or its analog, and
• (d) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence,
• wherein said assay composition is suitable for use in said assay based on said electrochemiluminescent phenomenon.

An assay kit containing reagents for use in a particle-based electrochemiluminescence binding assay for determining the presence or amount of an analyte of interest, comprising, in one or more vessels, the following:

• (a) at least one reagent substance selected from the group consisting of
• (i) added analyte of interest or an analog of the analyte of interest,
• (ii) a binding partner of the analyte of interest or its said analog, and
• (iii) a reactive component capable of binding with (i) or (ii), wherein one of said reagent substances is linked to a label compound capable of being induced to electrochemiluminesce,
• (b) a second reagent comprising a plurality of particles which:
• (i) have a surface capable of binding with the analyte or (ii) are attached to the analyte or its analog, and
• (c) a reductant that under oxidizing conditions interacts with the label compound to generate electrochemiluminescence,
• wherein said assay kit is suitable for use in said assay based on said electrochemiluminescent phenomenon.

A method for the detection and quantitation of an analyte of interest in a sample, which method comprises the steps of:

• (a) forming a composition comprising:
• (i) a sample,
• (ii) a first reagent comprising a binding partner of the analyte of interest, wherein said binding partner is linked to a label compound capable of being induced to electrochemiluminesce, and
• (iii) a second reagent capable of specifically binding with the analyte, said second reagent comprising particles selected from the group consisting of polymeric particles, inorganic particles, metallic particles, metal oxide particles, composite particles and mixtures thereof;
• (b) inducing the label compound to electrochemiluminesce;
• and
• (c) measuring luminescence emitted by the label compound to determine whether or not or in what amount, the analyte of interest is present in the sample.

A method for the detection and quantitation of an analyte of interest in a sample, which method comprises the steps of:

• (a) forming a composition comprising:
• (i) a sample,
• (ii) a first reagent comprising a binding partner of the analyte of interest, wherein said binding partner is linked to a label compound capable of being induced to electrochemiluminesce, and
• (iii) a second reagent linked to added analyte of interest or its analog, said second reagent comprising particles selected from the group consisting of polymeric particles, inorganic particles, metallic particles, metal oxide particles, composite particles and mixtures thereof;
• (b) inducing the label compound to electrochemiluminesce;
• and
• (c) measuring luminescence emitted by the label compound to determine whether or not, or in what amount, the analyte of interest is present in the sample.

A method for the detection and quantitation of an analyte of interest in a sample, which method comprises the steps of:

• (a) forming a composition comprising:
• (i) a sample,
• (ii) a first reagent comprising a binding partner of the analyte of interest, wherein said binding partner is linked to a label compound capable of being induced to electrochemiluminesce, and
• (iii) a second reagent linked to added analyte of interest or its analog, said second reagent comprising particles selected from the group consisting of polymeric particles, inorganic particles, metallic particles, metal oxide particles, composite particles and mixtures thereof;
• (b) inducing the label compound to electrochemiluminesce;
• and
• (c) measuring luminescence emitted by the label compound in the presence of said particles to determine whether or not or in what amount, the analyte of interest is present in the sample.