Process for detecting the presence or quantity of enzymatic activity in a sample

• US 7,163,796 B2
• Filed: 01/23/2004
• Issued: 01/16/2007
• Est. Priority Date: 03/12/2002
• Status: Expired due to Term

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: C/O ENZO Biochem Incorporated, Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Stavrianopoulos, Jannis G., Rabbani, Elazar
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US10/764,418
• Publication Number:
  US 20040176586A1
• Time in Patent Office: 1,089 Days
• Field of Search: 435/6, 435/7.1, 435/7.2, 536/26.6, 549/510, 436/172, 252/301.16, 252/186.42
• US Class Current: 435/7.1
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C09B 47/00 Porphines; Azaporphines non-dyeing compounds C07D487/22
• CPC Class Codes: C09B 57/00 Other synthetic dyes of known constitution
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

This invention provides for labeling reagents, labeled targets and processes for preparing labeling reagents. The labeling reagents can take the form of cyanine dyes, xanthene dyes, porphyrin dyes, coumarin dyes or composite dyes. These labeling reagents are useful for labeling probes or targets, including nucleic acids and proteins. These reagents can be usefully applied to protein and nucleic acid probe based assays. They are also applicable to real-time detection processes.

Claims

1. A process for detecting the presence or quantity of enzymatic activity of interest in a sample, said process comprising the steps of:

(a) providing;

(i) said sample suspected of containing enzymatic activity;

(ii) a chemiluminescent reagent having the structure;

2. The process of claim 1, wherein in said providing step (a) Q in said chemiluminescent reagent (ii) comprises an adamantyl group.

3. The process of claim 1, wherein in said providing step (a) R₂ in said chemiluminescent reagent (ii) comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.

4. The process of claim 3, wherein said substituted aliphatic group comprises halogen or sulfonates.

5. The process of claim 1, wherein said enzymatic converting step (c) is carried out by a substrate comprising glucose, xylose, fucose, amino acids, or esters of phosphates, carboxylic acids or fatty acids.

6. The process of claim 1, wherein said enzymatic activity of interest comprises an amidase, a trypsin or a chymotrypsin.

7. The process of claim 1, further comprising the step of forming an intermediate five- or six-membered ring comprising a linkage between said G₁ and G₂ in said chemiluminescent reagent (ii).

8. The process of claim 1, wherein any of said steps (a) through (d) are carried out in liquid phase or mixed phase.

9. The process of claim 1, wherein said enzymatic activity of interest is dependent upon the presence or quantity of another compound.

10. The process of claim 9, wherein said another compound comprises an RNA or DNA probe.

11. A process for detecting the presence or quantity of enzymatic activity of interest in a sample, said process comprising the steps of:

(a) providing;

(i) said sample suspected of containing enzymatic activity;

(ii) a chemiluminescent reagent having the structure;

12. The process of claim 11, wherein in said providing step (a) Q In said chemiluminescent reagent (ii) comprises an adamantyl group.

13. The process of claim 11, wherein in said providing step (a) R in the chemiluminescent reagent (ii) comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.

14. The process of claim 13, wherein said substituted aliphatic group comprises halogen or sulfonate.

15. The process of claim 11, wherein said providing step (a) R in the chemiluminescent reagent (ii) comprises a reactive site comprising an oxygen, a nitrogen or a sulfur atom.

16. The process of claim 11, wherein said step of enzymatically converting (c) is carried out by an enzyme comprising an oxidase or reductase.

17. The process of claim 11, wherein any of said steps (a) through (d) are carried out in liquid phase or mixed phase.

18. The process of claim 11, wherein said enzymatic activity of interest is dependent upon the presence or quantity of another compound.

19. The process of claim 18, wherein said another compound comprises an RNA or DNA probe.

20. A process for detecting the presence or quantity of enzymatic activity of interest in a sample, said enzymatic activity being dependent upon the presence or quantity of another compound, said process comprising the steps of:

(a) providing;

(i) said sample suspected of containing enzymatic activity that is dependent upon the presence or quantity of said another compound;

(ii) a chemiluminescent reagent having the structure;

21. A process for detecting the presence or quantity of enzymatic activity of interest in a sample, said enzymatic activity being dependent upon the presence or quantity of an RNA or DNA probe, said process comprising the steps of:

(a) providing;

(i) said sample suspected of containing enzymatic activity that is dependent upon the presence or quantity of said RNA or DNA probe;

(ii) a chemiluminescent reagent having the structure;