Claims for Patent: 5,145,772
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Summary for Patent: 5,145,772
Title: | Chemiluminescence enhancement of enzyme-activated decomposition of enzymatically cleavable chemiluminescent 1,2-dioxetanes |
Abstract: | Water soluble naturally-occurring and synthetic enhancer substances, generally macromolecular in nature, for example globular proteins that include hydrophobic regions such as bovine serum albumin, and polymeric quaternary ammonium salts such as poly(vinylbenzyltrimethylammonium chloride), which have the ability to inhibit light-emitting fluorophores resulting from the decomposition of chemiluminescent compounds from releasing energy through non-light emitting pathways, are disclosed as permitting the stabilization, and hence increasing the light intensity, of such light-emitting fluorophores in aqueous media as compared to the intensity of the light emitted by the same quantities of such fluorophores in aqueous media in the absence of such enhancer substances. Any chemiluminescent enzymatically cleavable 1,2-dioxetane, for example 3-(2\'-spiroadamantane)-4-methoxy-(3\"-phosphoryloxy)phenyl-1,2-dioxetane disodium salt, can be used. Auxiliary fluorophores, for example fluorescein and derivatized fluoresceins, that accept energy from fluorophores produced by decomposition of a chemiluminescent compound and in turn emit detectable energy, can also be present. Such enhancer substance/chemiluminescent compound compositions are useful in detecting the presence or determining the concentration of chemical or biological substances in immunoassays, chemical assays and nucleic acid probe assays, and in chemical/physical probe procedures for studying the microstructures of macromolecules. |
Inventor(s): | Voyta; John C. (North Reading, MA), Edwards; Brooks (Cambridge, MA), Bronstein; Irena Y. (Newton, MA), McGrath; Patricia (Cambridge, MA) |
Assignee: | Tropix, Inc. (Bedford, MA) |
Application Number: | 07/767,566 |
Patent Claims: | 1. In a process carried out in an aqueous medium in which electromagnetic energy released by the enzyme-activated decomposition of an enzymatically cleavable chemiluminescent
1,2-dioxetane to produce an anionic electromagnetic energy-emitting fluorophore is detected to determine the presence, concentration or structure of an analyte, the improvement comprising carrying out the process in the presence of an amount of an
admixed water soluble macromolecular enhancer substance sufficient to enhance the intensity of any released detectable electromagnetic energy over the intensity of any detectable electromagnetic energy released in the absence of the enhancer substance,
wherein the enhancer substance is itself incapable of energy transfer.
2. The process of claim 1 in which the macromolecular enhancer substance has the ability to inhibit the fluorophore from releasing energy through non-light emitting pathways. 3. The process of claim 2 in which the macromolecular enhancer substrate comprises a naturally-occurring or synthetic macromolecular substance that can provide a hydrophobic microenvironment for the fluorophore. 4. The process of claim 3 in which the macromolecular enhancer substance comprises a naturally-occurring macromolecular substance. 5. The process of claim 4 in which the macromolecular substance is a globular protein that includes hydrophobic regions. 6. The process of claim 5 in which the globular protein is a mammalian serum albumin. 7. The process of claim 6 in which the serum albumin is bovine serum albumin. 8. The process of claim 6 in which the serum albumin is human serum albumin. 9. The process of claim 5 in which the globular protein is mammalian IgG. 10. The process of claim 5 in which the globular protein is Protein A. 11. The process of claim 3 in which the macromolecular enhancer substance comprises a synthetic macromolecular substance. 12. The process of claim 11 in which the macromolecular enhancer substance comprises an oligomeric or polymeric quaternary ammonium salt. 13. The process of claim 12 in which the quaternary ammonium salt is a poly(vinylaryl quaternary ammonium salt). 14. The process of claim 13 in which the poly(vinylaryl quaternary ammonium salt) is poly(vinylbenzyltrimethylammonium chloride). 15. The process of claim 13 in which the poly(vinylaryl quaternary ammonium salt) is poly(vinylbenzyl(benzyldimethylammonium chloride)). 16. The process of claim 1 in which the 1,2-dioxetane is a 3-(2'spiroadamantane)-4-methoxy-(3"-phosphoryloxy)phenyl-1,2-dioxetane salt. 17. The process of claim 1 further comprising carrying out the process in the presence of an admixed auxiliary fluorophore extraneous to the electromagnetic energy-emitting fluorophore produced by the decomposition of the chemiluminescent 1,2-dioxetane, the auxiliary fluorophore being capable of accepting energy from the electromagnetic energy-emitting fluorophore to in turn emit detectable energy. 18. The process of claim 17 in which the auxiliary fluorophore is admixed with the chemiluminescent 1,2-dioxetane and the macromolecular enhancer substance. 19. The process of claim 18 in which the auxiliary fluorophore is a fluorescein. 20. The process of claim 17 in which the auxiliary fluorophore is covalently bonded to the portion of the chemiluminescent 1.2-dioxetane that, upon decomposition, produces the electromagnetic energy-emitting fluorophore. 21. The process of claim 20 in which the auxiliary fluorophore is a fluorescein. 22. The process of claim 17 in which the auxiliary fluorophore is covalently bonded to the macromolecular enhancer substance. 23. The process of claim 22 in which the auxiliary fluorophore is a derivatized fluorescein capable of establishing a covalent bond with the macromolecular enhancer substance. 24. The process of any one of claims 1, 17 or 22 in which the process carried out is a step in an immunoassay. 25. The process of claim 24 in which the immunoassay is for the detection of a specific binding pair comprising an antigen and an antibody, there being bonded to either or both of the antigen and the antibody a label. 26. The process of claim 25 in which the label used in the immunoassay is an enzyme. 27. The process of claim 24 in which the immunoassay is for the detection of an enzyme. 28. The process of claim 24 in which the immunoassay is for the detection of a hormone. 29. The process of any one of claims 1, 17 or 22 in which the process carried out is a step in a chemical assay. 30. The process of claim 29 in which the chemical substance is cholesterol. 31. The process of claim 29 in which the chemical substance is glucose. 32. The process of any one of claims 1, 17 or 22 in which the process carried out is a nucleic acid probe assay. 33. The process of claim 32 in which the nucleic acid probe assay is for the detection of a virus. 34. The process of claim 32 in which the nucleic acid probe assay is for the detection of a bacteria. 35. The process of any one of claims 1, 17 or 22 in which the process carried out is a histocompatibility assay. 36. The process of any one of claims 1, 17 or 22 in which the process carried out is a technique for studying the microstructure of a macromolecule. |
Details for Patent 5,145,772
Applicant | Tradename | Biologic Ingredient | Dosage Form | BLA | Approval Date | Patent No. | Expiredate |
---|---|---|---|---|---|---|---|
Grifols Therapeutics Llc | ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5 | albumin (human) | For Injection | 101138 | 10/21/1942 | ⤷ Try a Trial | 2009-09-08 |
Baxalta Us Inc. | BUMINATE, FLEXBUMIN | albumin (human) | Injection | 101452 | 03/03/1954 | ⤷ Try a Trial | 2009-09-08 |
Csl Behring Ag | ALBURX | albumin (human) | Injection | 102366 | 07/23/1976 | ⤷ Try a Trial | 2009-09-08 |
Grifols Biologicals Llc | ALBUTEIN | albumin (human) | Injection | 102478 | 08/15/1978 | ⤷ Try a Trial | 2009-09-08 |
Instituto Grifols, S.a. | HUMAN ALBUMIN GRIFOLS | albumin (human) | Injection | 103352 | 02/17/1995 | ⤷ Try a Trial | 2009-09-08 |
Instituto Grifols, S.a. | HUMAN ALBUMIN GRIFOLS | albumin (human) | Injection | 103352 | 06/11/2003 | ⤷ Try a Trial | 2009-09-08 |
>Applicant | >Tradename | >Biologic Ingredient | >Dosage Form | >BLA | >Approval Date | >Patent No. | >Expiredate |
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