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| Method of fermenting brewer's wort |
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Use of oxidase enzyme systems in chemiluminescent assays
| Details |
Inventors: Baret, Alain;
Assignee: Canberra Industries, Inc. (Meriden, CT)
Primary Examiner: Kepplinger; Esther L.
Assistant Examiner: Wolski; Susan C.
Attorney, Agent or Firm: Arnold, White & Durkee
A xanthine oxidase enzyme system to provide long lived entities capable of being recognized by a chemiluminescent reagent is disclosed. In the examples provided, a specific binding pair ligand or analyte is coupled with xanthine oxidase, either directly or via a streptavidin bridge. Thereafter, the presence of an analyte can be determined by a chemiluminescent emission upon addition of a signal reagent comprising hypoxanthine, iron EDTA complex and luminol dissolved in barbital buffer. The resulting chemiluminescent signal is stable and detectable for many hours after initiation. The chemiluminescent xanthine oxidase system is particularly useful for immunoassays and DNA probe analysis. |
|
DETAILED DESCRIPTION What is claimed is: 1.
A kit for use in a chemiluminescent assay to detect an oligonucleotide analyte, comprising the following reagents in separate containers: a) a solid support for depositing the analyte; b) a conjugate of a complementary nucleotide binding partner to the analyte and an oxidase, said oxidase capable of producing both H.
sub.
2 O.
sub.
2 and superoxide anion; and c) a signal reagent comprising at a pH ranging from about 9.
5 to about 10.
5, a substrate for said oxidase, a chemiluminescent reagent, and an iron EDTA enhancer.
2.
The kit of claim I wherein the oxidase is xanthine oxidase, choline oxidase, fumarate oxidase or sarcosine oxidase.
3.
The kit of claim 1 wherein the oxidase is xanthine oxidase.
4.
The kit of claim 3 wherein the oxidase substrate is xanthine, hypoxanthine, acetaldehyde or purine.
5.
The kit of claim 3 wherein the oxidase substrate is hypoxanthine.
6.
The kit of claim 1 wherein the chemiluminescent reagent is luminol.
7.
The kit of claim I wherein the signal reagent is maintained in barbital, carbonate or borate buffer.
8.
The kit of claim I wherein the oxidase is xanthine oxidase, and wherein the signal reagent comprises hypoxanthine, luminol and iron EDTA enhancer.
9.
A kit for use in a chemiluminescent assay to detect an oligonucleotide analyte, comprising the following reagents in separate containers: a) a solid support for depositing the analyte; b) a first conjugate of a biotin compound and a complementary nucleotide binding partner to the analyte; c) a second conjugate of an avidin compound and an oxidase, said oxidase capable of producing both H.
sub.
2 O.
sub.
2 and superoxide anion; and d) a signal reagent comprising at a pH ranging from about 9.
5 to about 10.
5, a substrate for said oxidase, a chemiluminescent reagent, and an iron EDTA enhancer.
10.
The kit of claim 9 wherein the oxidase is xanthine oxidase, choline oxidase, fumarate oxidase or sarcosine oxidase.
11.
The kit of claim 9 wherein the oxidase is xanthine oxidase 12
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Molecular Biology 
