| 主权项 |
1. A solution phase assay method for an analyte in a sample, the solution phase assay method comprising:
a) forming an aqueous reaction mixture comprising the following components:
the sample,a chemiluminescent-labeled specific binding partner comprising a chemiluminescent label compound connected directly or indirectly to a first specific binding partner, wherein the chemiluminescent label compound is a compound of the formula wherein designates the point of attachment of the chemiluminescent label to the first specific binding partner, wherein R1 and R2 are independently selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl groups of 1-20 carbon atoms, wherein when R1 or R2 is a substituted group, it can be substituted with 1-3 groups selected from the group consisting of carbonyl groups, carboxyl groups, tri(C1-C8 alkyl)silyl groups, a SO3− group, a OSO3−2 group, glycosyl groups, a PO3− group, a OPO3−2 group, halogen atoms, a hydroxyl group, a thiol group, amino groups, a C(═O)NHNH2 group, quaternary ammonium groups, and quaternary phosphonium groups, wherein R3 is selected from the group consisting of alkyl, substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl groups of 1-20 carbon atoms, phenyl, substituted or unsubstituted benzyl groups, alkoxyalkyl, carboxyalkyl and alkylsulfonic acid groups, wherein when R3 is a substituted group, it can be substituted with 1-3 groups selected from the group consisting of carbonyl groups, carboxyl groups, tri(C1-C8 alkyl)silyl groups, a SO3− group, a OSO3−2 group, glycosyl groups, a PO3− group, a OPO3−2 group, halogen atoms, a hydroxyl group, a thiol group, amino groups, quaternary ammonium groups, and quaternary phosphonium groups,
an activator-labeled specific binding partner comprising an activator label compound having peroxidase activity connected directly or indirectly to a second specific binding partner, anda selective signal inhibiting agent selected from the group consisting of L-ascorbic acid, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, 2-aminophenol, 3-amino-L-tyrosine, 4-chlorocatechol, phenoxazine, 2-bromobenzene-1,4-diol, 5,6-isopropylidene ascorbic acid, and ascorbic acid 6-palmitate, by adding the components, in any order or concurrently, to an aqueous solution,
wherein all of the components are soluble in the aqueous solution and none of the components are immobilized to a solid support, andwherein the chemiluminescent-labeled specific binding partner and the activator-labeled specific binding partner bind to the analyte present in the sample to form a binding complex in the aqueous solution; and b) adding to the aqueous reaction mixture a trigger solution comprising a peroxide compound, wherein the trigger solution releases a detectable chemiluminescent signal in the presence of the selective signal inhibiting agent that is correlated to the amount of the analyte-bound chemiluminescent-labeled specific binding partner and the analyte-bound activator-labeled specific binding partner in the aqueous reaction mixture, and wherein the selective signal inhibiting agent causes the ratio of signal produced by reaction between the chemiluminescent label compound and the activator label compound in the complex with the analyte to exceed the signal from reaction between the chemiluminescent label compound and the activator label compound when not in such a complex. |
