Electrochemiluminescent reaction utilizing amine-derived reductant
DC CAFCFirst Claim
1. An electrochemiluminescent assay method of determining the presence or absence of or quantitating an analyte of interest, which consists essentially of the steps of:
- (a) forming a composition consisting essentially of(i) a sample to be tested for said analyte of interest;
(ii) a specific binding component capable of specifically binding with said analyte to form a binding product in the event said analyte is present in said sample;
(iii) a label reagent comprising a chelate of a transition metal or a rare earth metal which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, and further comprising a species linked to said chelate which species is said analyte, an analog thereof, said specific binding component as recited in subparagraph (ii), a binding partner of said analyte, analog or specific binding component, or a binding partner capable of forming part of a combination of binding partners wherein at least one of the binding partners in such combination is capable of binding with said analyte, said analog, or said specific binding component;
(iv) an amine which, when oxidized, forms a reducing agent, said amine being selected from the group consisting of(x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines,(y) the amines defined in subparagraph (a)(iv)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups and heterocyclic groups, and(z) protonated salts of the amines defined in subparagraph (a)(iv)(x) and protonated salts of the substituted amines defined in subparagraph (a)(iv)(y); and
(v) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized;
(b) subjecting said composition to conditions sufficient for said specific binding to occur in the event said analyte is present in said sample;
(c) thereafter oxidizing the chelate and the amine such that an interaction between resulting oxidized products produces electrochemiluminescence; and
(d) detecting luminescence emitted as a result of the electrochemiluminescence of said label reagent to determine the presence or absence, or the amount, of said analyte.
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Abstract
Electrochemiluminescent assay methods for determining an analyte of interest, which methods include forming one or more compositions which include, in the aggregate, (a) a sample to be tested for the analyte of interest, (b) a component capable of specifically binding with the analyte, (c) a label reagent which, when oxidized, is capable of electrochemiluminescence, (d) an amine which, when oxidized, forms a reducing agent, and (e) an electrolyte solution; subjecting such a composition containing the sample to conditions sufficient for specific binding to occur between the analyte of interest, if present, and one or more of the other components in the composition; thereafter, a composition containing the label reagent and amine, and reflecting the outcome of applying the aforementioned binding conditions, is exposed to conditions, such that both the label reagent and the amine are oxidized, the amine forms a reducing agent which interacts with the label reagent, electrochemiluminescence occurs, and the luminescence emitted from the label reagent is used to determine the analyte of interest.
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Citations
61 Claims
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1. An electrochemiluminescent assay method of determining the presence or absence of or quantitating an analyte of interest, which consists essentially of the steps of:
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(a) forming a composition consisting essentially of (i) a sample to be tested for said analyte of interest; (ii) a specific binding component capable of specifically binding with said analyte to form a binding product in the event said analyte is present in said sample; (iii) a label reagent comprising a chelate of a transition metal or a rare earth metal which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, and further comprising a species linked to said chelate which species is said analyte, an analog thereof, said specific binding component as recited in subparagraph (ii), a binding partner of said analyte, analog or specific binding component, or a binding partner capable of forming part of a combination of binding partners wherein at least one of the binding partners in such combination is capable of binding with said analyte, said analog, or said specific binding component; (iv) an amine which, when oxidized, forms a reducing agent, said amine being selected from the group consisting of (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (a)(iv)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (a)(iv)(x) and protonated salts of the substituted amines defined in subparagraph (a)(iv)(y); and (v) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (b) subjecting said composition to conditions sufficient for said specific binding to occur in the event said analyte is present in said sample; (c) thereafter oxidizing the chelate and the amine such that an interaction between resulting oxidized products produces electrochemiluminescence; and (d) detecting luminescence emitted as a result of the electrochemiluminescence of said label reagent to determine the presence or absence, or the amount, of said analyte. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 30)
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18. An electrochemiluminescent assay method of determining the presence or absence of or quantitating an analyte of interest, which consists essentially of the steps of:
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(a) forming a first composition consisting essentially of a sample to be tested for said analyte of interest and a specific binding component capable of specifically binding with said analyte to form a binding product in the event said analyte is present in said sample; (b) subjecting said first composition to conditions sufficient for said specific binding to occur in the event said analyte is present in said sample; (c) forming a second composition consisting essentially of (i) said specific binding component, or said product of said specific binding, or both; (ii) a label reagent comprising a chelate of a transition metal or a rare earth metal which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, and further comprising a species linked to said chelate which species is said analyte, an analog thereof, said specific binding component as recited in subparagraph (i), the product of said specific binding as recited in subparagraph (i), a binding partner of said analyte, analog, specific binding component, product of specific binding, or a binding partner which is capable of forming part of a combination of said binding partners wherein at least one of the binding partners in such combination is capable of binding with said analyte, said analog, said specific binding component, or said product of said specific binding; (iii) an amine which, when oxidized, forms a reducing agent, said amine being selected from the group consisting of; (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (c)(iii)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O), --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (c)(iii)(x), and protonated salts of the substituted amines defined in subparagraph (c)(iii)(y); and (iv) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (d) thereafter oxidizing the chelate and the amine such that an interaction between resulting oxidized products produces electrochemiluminescence; and (e) detecting luminescence emitted as a result of the electrochemiluminescence of said label reagent to determine the presence or absence, or the amount, of said analyte. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. A method of determining the presence or absence of or quantitating an analyte of interest by electrochemiluminescent assay, which consists essentially of:
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(1) forming a reaction mixture consisting essentially of (a) a sample to be tested for the analyte; (b) at least one substance which is (i) a predetermined amount of said analyte or an analog of the analyte, and (ii) a binding partner of the analyte or said analog thereof, or a binding partner which is capable of forming part of a combination of binding partners wherein at least one of the binding partners in such combination is capable of binding with said analyte or said analog, (c) a label reagent comprising a chelate of a transition metal or a rare earth metal, which reagent (i) is capable of specifically binding with the analyte or at least one substance present from among (b)(i) and(b)(ii), and (ii) when oxidized, is capable of being converted to an excited state which electrochemiluminesces; (d) an amine which, when oxidized, forms reducing agent, said amine being selected from the group consisting off (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, and (y) the amines defined in subparagraph (1)(d)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (1)(d)(x), and protonated salts of the substituted amines defined in subparagraph (1)(d)(y); and (e) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (2) subjecting said composition to conditions sufficient for said specific binding to occur in the event said analyte is present in said sample; (3) thereafter oxidizing the chelate and the amine such that an interaction between resulting oxidized products produces electrochemiluminescence; and (4) detecting luminescence emitted as a result of the electrochemiluminescence of said label reagent to determine the presence or absence of or the amount of said analyte. - View Dependent Claims (29, 31, 32, 33, 34)
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35. A system for electrochemiluminescent detection or quantitation of an analyte of interest in a sample, said system consisting essentially of
(a) at least one substance which is a predetermined amount of (i) said analyte or an analog of the analyte, and (ii) a binding partner of the analyte or said analog thereof, or a binding partner which is capable of forming part of a combination of binding partners wherein at least one of the binding partners in such combination is capable of binding with said analyte or said analog, said at least one substance being linked to a chelate of a transition metal or a rare earth metal, which chelate is capable of being converted to an excited state which electrochemiluminesces; -
(b) an amine which, when oxidized, forms a reducing agent, said amine being selected from the group consisting of (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (b)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (b)(x), and protonated salts of the substituted amines defined in subparagraph (b)(y); and (c) an electrolyte; (d) means for oxidizing both the chelate and the amine such that an interaction between resulting oxidized products produces electrochemiluminescence; and (e) means for detecting emitted luminescence to determine the presence or absence or the amount of said analyte in the sample. - View Dependent Claims (36, 37)
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38. An electochemiluminescent assay method for determining the presence of or quantitating an analyte of interest, said method comprising the steps of:
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(a) forming a complex comprising; (i) said analyte; (ii) a first specific binding partner of said analyte; and (iii) an electrochemiluminescent label comprising a chelate of a transition metal or a rare earth metal, which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, wherein said chelate is linked to said first specific binding partner; (b) forming a composition comprising; (i) said complex; (ii) an amine which forms a reducing agent upon oxidation, said amine being selected from the group consisting of; (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (b)(ii)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (b))(ii)(x) and protonated salts of the substituted amines defined in subparagraph (b)(ii)(y); and (iii) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (c) exposing said composition to electrochemical energy such that both the chelate and the amine are oxidized and an interaction between resulting oxidized products produces electrochemiluminescence, said electrochemiluminescence being related to the amount of said analyte; (d) detecting said electrochemiluminescence and thereby determining the presence or amount of said analyte. - View Dependent Claims (39, 40, 41, 42, 43)
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44. An electrochemiluminescent assay method for determining the presence of or quantitating an analyte of interest, said method comprising the steps of:
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(a) forming a complex comprising; (i) an analog of said analyte; (ii) a first specific binding partner of said analyte and said analog; and (iii) an electrochemiluminescent label comprising a chelate of a transition metal or a rare earth metal, which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, wherein said chelate is linked to said analog and said analog competes for binding with said analyte for said first specific binding partner; (b) forming a composition comprising; (i) said complex; (ii) said analyte; (iii) an amine which forms a reducing agent upon oxidation, said amine being selected from the group consisting of; (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (b)(iii)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (b)(iii)(x) and protonated salts of the substituted amines defined in subparagraph (b)(iii)(y); and (iii) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (c) exposing the composition to electrochemical energy such that both the chelate and the amine are oxidized and an interaction between resulting oxidized products produces electrochemiluminescence, said electrochemiluminescence being related to the amount of said analyte present; and (d) detecting said electrochemiluminescence and thereby determining the presence or amount of said analyte. - View Dependent Claims (45, 46, 47, 48, 49)
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50. An electrochemiluminescent assay method for determining the presence of or quantitating an analyte of interest, said method comprising the steps of:
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(a) forming a first composition comprising; (i) said analyte; (ii) a first specific binding partner of said analyte; and (iii) an electrochemiluminescent label comprising a chelate of a transition metal or a rare earth metal, which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, wherein said chelate is linked to said first specific binding partner; (b) forming a second composition comprising; (i) said first composition; (ii) an amine which forms a reducing agent upon oxidation, said amine being selected from the group consisting of; (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (b)(ii)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (b)(ii)(x) and protonated salts of the substituted amines defined in subparagraph (b)(ii)(y); and (iii) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (c) exposing said second composition to electrochemical energy such that both the chelate and the amine are oxidized and an interaction between resulting oxidized products produces electrochemiluminescence, said electrochemiluminescence being related to the amount of said analyte present; (d) detecting said electrochemiluminescence and thereby determining the presence or amount of said analyte. - View Dependent Claims (51, 52, 53, 54, 55)
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56. An electrochemiluminescent assay method for determining the presence or absence of or quantitating an analyte of interest, said method comprising the steps of:
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(a) forming a first composition comprising; (i) an analog of said analyte; (ii) a first specific binding partner of said analyte and said analog; and (iii) an electrochemiluminescent label comprising a chelate of a transition metal or a rare earth metal, which chelate, when oxidized, is capable of being converted to an excited state which electrochemiluminesces, wherein said chelate is linked to said analog and said analog and said first specific binding partner bind to one another if said analyte is present, such that said analyte competes for binding with said analog; (b) forming a second composition comprising; (i) said first composition; (ii) said analyte; (iii) an amine which forms a reducing agent upon oxidation, said amine being selected from the group consisting of; (x) aliphatic amines, aromatic amines, diamines, polyamines, and heterocyclic amines, (y) the amines defined in subparagraph (b)(iii)(x) substituted by one or more substituents selected from the group consisting of --OH, alkyl, chloro, fluoro, bromo, iodo, --SO3, aryl, --SH, --C(O)--H, --C(O)--OH, ester groups, ether groups, alkenyl, alkynyl, --C(O)--, --N2+, cyano, epoxide groups, and heterocyclic groups, and (z) protonated salts of the amines defined in subparagraph (b)(iii)(x) and protonated salts of the substituted amines defined in subparagraph (b)(iii)(y); and (iv) an electrolyte solution which functions as a medium in which said chelate and said amine can be oxidized; (c) exposing the second composition to electrochemical energy such that both the chelate and the amine are oxidized and an interaction between resulting oxidized products produces electochemiluminescence, said electrochemiluminescence being related to the amount of said analyte present; and (d) detecting said electrochemiluminescence and thereby determining the presence or amount of said analyte. - View Dependent Claims (57, 58, 59, 60, 61)
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Specification