JP2005520175A5 - - Google Patents

Claims (13)

In a method for detecting at least first and second target molecules in a sample,
(A) providing a solid support comprising a conductive redox reaction detection electrode having at least a first capture binding ligand;
(B) the first target molecule forms a first assay complex with one capture binding ligand, and the second target molecule forms a second assay complex with one capture binding ligand. Contacting the sample with the electrode under the conditions of: wherein the first assay complex comprises a first preselected label having a first redox potential and the second assay. The complex includes a second preselected label having a second redox potential;
(C) the assay complex, (i) oxidizing the first preselected label in a first redox reaction and regenerating the reduced form of the first transition metal complex in a catalytic reaction. (Ii) the second transition that oxidizes the second preselected label in a second redox reaction and regenerates the reduced form of the second transition metal complex in a catalytic reaction Contacting with the metal complex;
(D) detecting the presence of the target analyte by detecting the first and second oxidation-reduction reactions.   The sample includes a third target molecule that forms a third assay complex with one capture binding ligand, and the third assay complex is selected from the third preselected in a third redox reaction. The method of claim 1, wherein the method comprises the third transition metal complex that oxidizes the labeled and regenerates the reduced form of the third transition metal complex in a catalytic reaction.   The sample includes a fourth target molecule that forms a fourth assay complex with one capture binding ligand, and the fourth assay complex is selected in the fourth pre-selected reaction in a fourth redox reaction. 3. The method of claim 2, comprising the fourth transition metal complex that oxidizes the labeled and regenerates the fourth transition metal complex in a catalytic reaction.   4. The method according to any one of claims 1 to 3, wherein the first capture binding ligand binds the first, second, third and fourth target molecules.   The electrode includes one or more additional capture binding ligands, each of the first, second, third and fourth target molecules binding to a different capture binding ligand; 4. A method according to any one of claims 1 to 3.   6. The method of any one of claims 1-5, wherein each of the assay complexes comprises one capture binding ligand and one target analyte.   Each of said assay complexes comprises one capture binding ligand, one target analyte and one label binding ligand, each of said binding ligands comprising a corresponding said preselected label. 6. The method according to any one of 1 to 5.   8. A method according to any one of claims 1 to 7, wherein one of the target molecules is a nucleic acid and another one of the target molecules is a protein.   The method according to any one of claims 1 to 7, wherein all of the target molecules are nucleic acids.   The method of claim 9, wherein the method comprises amplifying the nucleic acid prior to the contacting step. At least one of the transition metal complexes includes Ru (bpy) ₃ ²⁺ , Ru (Me ₂ -bpy) ₃ ²⁺ , Ru (Me ₂ -phen) ₃ ²⁺ , Fe (bpy) ₃ ²⁺ , 11. Any one of claims 1 to 10 selected from the group consisting of Fe (5-Cl-phen) ₃ ²⁺ , Os (5-Cl-phen) ₃ ²⁺ , and ReO ₂ (py) ₄ ^1+. The method according to claim 1.   At least one of the preselected bases is from adenine, 7-deazaadenine, guanine, 6-mercaptoguanine, 8-oxoguanine, isoguanine, 7-deazaguanine, 1-hydroxyisoguanine, and 8-bromoguanine. 11. A method according to any one of claims 1 to 10, wherein the method is selected from the group consisting of:   12. A method according to any one of the preceding claims, wherein the solid support comprises an array of a plurality of the electrodes.