Electrochemiluminescent reaction utilizing amine-derived reductant

  • US 6,165,729 A
  • Filed: 04/15/1994
  • Issued: 12/26/2000
  • Est. Priority Date: 04/30/1986
  • Status: Expired due to Term
First Claim
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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:

  • (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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