Assay
Method of assaying for an analyte in a sample, and kits for performing the assay.
1. A kit for assaying for an analyte, said kit comprising:
a first chemiluminescent compound, wherein said first chemiluminescent compound is an unbound chemiluminescent substrate;
a chemiluminescent-labeled specific binding partner comprising a second chemiluminescent compound, wherein the second chemiluminescent compound is a chemiluminescent label irreversibly bound to a first specific binding partner, the chemiluminescent-labeled specific binding partner being capable of binding configured to bind to the analyte to form an analyte-bound chemiluminescent labeled specific binding complex,
an activator-labeled specific binding partner comprising an activator label irreversibly bound to a second specific binding partner, and
a selective signal inhibiting agent; and
a trigger solution capable of producing configured to produce a background signal from the unbound chemiluminescent substrate and, in the presence of analyte, producing a detectable analyte signal correlated to an amount of analyte in a sample.
2. The kit of claim 1 , wherein the trigger solution further comprises an enhancer selected from phenol compounds, aromatic amines, benzoxazoles, hydroxybenzothiazoles, aryl boronic acids, and mixtures of any of the foregoing.
3. The kit of claim 1 , wherein the unbound chemiluminescent substrate is selected from a group consisting of luminol, isoluminol, lophine, acridinium esters such as lucigenin and acridan, phthalhydrazides such as 2,3-dihydro-1,4-phthalazinedione, luciferin, and 1,2 dioxyetane containing compounds such as dione, 3-(2′-spiroadamantane)-4-methoxy-4-(3″-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD), and, 3-(2′-spiroadamantane)-4-methoxy-4-(3″-beta-D′-galactopyrano-yloxy)phenyl-1,2-dioxetane (AMPGD), disodium 3-(4-methoxyspiro {1,2-dioxetane-3,2′-(5′-chloro)tricyclo[3.3.1.13,7]decan}-4-yl)phenyl phosphate, and 1,2-dioxetanedione, adamantylidene-adamantyl-1,2-dioxetane, particularly luminol or isoluminol, and more particularly luminol.
4. The kit of claim 1 , wherein said unbound chemiluminescent substrate comprises luminol.
5. The kit of claim 1 , wherein the wavelength of maximum luminescence intensity of the unbound chemiluminescent substrate is different from the wavelength of maximum luminescence intensity of the chemiluminescent label.