Electrochemiluminescent assays

• US 6,316,607 B1
• Filed: 06/07/1995
• Issued: 11/13/2001
• Est. Priority Date: 04/30/1986
• Status: Expired due to Term

• Current Assignee: BioVeris Corporation (Roche Holding AG)
• Original Assignee: IGEN International, Inc. (Roche Holding AG)
• Inventors: Dressick, Walter J., Della Ciana, Leopoldo, Powell, Michael J., Mied, Paul A., Feng, Peter, Massey, Richard J., Poonian, Mohindar S.
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US08/472,425
• Time in Patent Office: 2,351 Days
• Field of Search: 435/6, 536/22.1, 536/23.1, 536/25.3, 514/184, 514/258, 514/277
• US Class Current: 536/22.1
• CPC Class Codes: C07D 473/08 with methyl radicals in positions 1 and 3, e.g. theophylline
• CPC Class Codes: C07F 15/0026 without a metal-carbon linkage
• CPC Class Codes: C07F 15/0053 without a metal-carbon linkage
• 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: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors

Abstract

Qualitative and quantitative electrochemiluminescent assays for analytes of interest present in multicomponent liquids are provided. These methods comprise contacting a sample with a reagent labeled with an electrochemiluminescent chemical moiety and capable of combining with the analyte of interest, exposing the resulting sample to electrochemical energy and detecting electromagnetic radiation emitted by the electrochemiluminescent chemical moiety.

Claims

1. A composition of matter having the structure:

2. A composition of matter of claim 1 wherein X—

Y—

Z comprises

3. A composition of matter of claim 1, wherein X—

Y—

Z comprises

20. A composition of matter of claim 1, wherein X is theophylline.

21. A composition of matter of claim 1, wherein X is digoxigenin.

22. A composition of matter of claim 1, wherein X is a peptide derived from human chorionic gonadotropin.

4. A composition of matter having the structure X—

CH═

CH—

CO—

NH—

(CH₂)_n—

NH—

CO—

(CH₂)_m—

Z, wherein X is thymidine attached to CH at carbon 5 of thymidine, n is 7, m is 3 and Z is bis(2,2′

-bipyridine)(4-(butan-1-al)-4-methyl-2,2′

-bipyridine)ruthenium (II).

23. A composition of matter as defined in claim 4, wherein the thymidine nucleotide is a 3′

terminal nucleotide attached to the 5′

→

3′

nucleotide sequence

5. A composition of matter having the structure

24. A composition of matter of claim 5, wherein Y is attached to the carbon at position 8 of T.

25. A composition of matter of claim 24, wherein Y has the structure

26. A composition of matter of claim 25, wherein m is 3 and n is 4.

27. A composition of matter of claim 25, wherein m and n are both 3.

28. A composition of matter of claim 24, wherein Y has the structure

and wherein m, n and r represent an integer, which may be the same of different, greater than or equal to 1.

29. A composition of matter of claim 28, wherein m is 1, n is 1 and r is 4.

30. A composition of matter of claim 24, wherein Y has the structure

and wherein m, n and r represent an integer, which may be the same or different, greater than or equal to 1.

31. A composition of matter of claim 30, wherein m is 1, n is 1 and r is 4.

32. A composition of matter of claim 5, wherein Y is attached to the nitrogen at position 7 of T.

33. A composition of matter of claim 32, wherein Y has the structure

34. A composition of matter of claim 33, wherein n is 4.

6. A compound which is the product of a linkage reaction between X and Z, wherein

X represents one or more analytes of interest or analogues of the analytes of interest, wherein the analytes are the same or not the same and selected from the group consisting of a nucleotide, amino acid, antibody, theophylline, digoxin, human chorionic gonadotropin, whole cell, subcellular particle, virus, prion, viroid, nucleic acid, protein, lipoprotein, lipopolysaccharide, glycoprotein, peptide, hormone, pharmacological agent, nonbiological polymer, synthetic organic molecule and organometallic molecule;

and

Z represents a compound selected from the group consisting of;

(1);

wherein A is —

CH₂—

(CH₂)_n—

CHO, —

CH₂—

(CH₂)_n—

COOH, —

CH₂—

(CH₂)_n—

NH₂, —

CH₂—

(CH₂)_n—

Br, and —

CH₂—

(CH₂)_n—

CO₂—

NHS, wherein NHS represents N-hydroxysuccinimide;

(2);

wherein n is an integer, the linkage reaction between X and Z results in a linkage —

(CH₂)_m—

between X and Z, and m is an integer greater than or equal to 1.

7. The compound defined in claim 6, wherein X is theophylline

8. The compound defined in claim 6, wherein X is digoxigenin.

9. The compound defined in claim 6, wherein X is a peptide derived from human chorionic gonadotropin.

10. A compound having the structure:

11. A composition of matter of claim 10, wherein X is theophylline.

12. A composition of matter of claim 10, wherein X is digoxigenin.

13. A composition of matter of claim 10, wherein X is a peptide derived from human chorionic gonadotropin.

14. A compound having the structure:

wherein G is a moiety that reacts with a species selected from the group consisting of a nucleotide, amino acid, antibody, theophylline, digoxin, human chorionic gonadotropin, whole cell, subcellular particle, virus, prion, viroid, nucleic acid, protein, lipoprotein, lipopolysaccharide, glycoprotein, peptide, hormone, pharmacological agent, nonbiological polymer, synthetic organic molecule and an organometallic molecule, wherein G and said species react to form a covalent linkage between said compound and said species;

R is H, alkyl or CO₂Q, wherein Q is H or alkyl;

M is Ru or Os;

n is an integer; and

D is H or alkyl.

16. The compound defined in claim 14, wherein G is selected from the group consisting of:

wherein m is an integer.

15. A compound having the structure:

wherein G is a moiety that reacts with a species selected from the group consisting of a nucleotide, amino acid, antibody, theophylline, digoxin, human chorionic gonadotropin, whole cell, subcellular particle, virus, prion, viroid, nucleic acid, protein, lipoprotein, lipopolysaccharide, glycoprotein, peptide, hormone, pharmacological agent, nonbiological polymer, synthetic organic molecule and an organometallic molecule, wherein G and said species react to form a covalent linkage between said compound and said species;

L is a bipyridyl or substituted bipyridyl bidentate ligand;

M is Ru or Os;

n is an integer; and

D is H or alkyl.

17. The compound defined in claim 15, wherein G is selected from the group consisting of:

—

CHO, —

CO₂H, —

NH₂,

18. A composition of matter having the structure:

wherein G is a moiety that reacts with a species selected from the group consisting of a nucleotide, amino acid, antibody, theophylline, digoxin, human chorionic gonadotropin, whole cell, subcellular particle, virus, prion, viroid, nucleic acid, protein, lipoprotein, lipopolysaccharide, glycoprotein, peptide, hormone, pharmacological agent, nonbiological polymer, synthetic organic molecule and an organometallic molecule, wherein G and said species react to form a covalent linkage between said compound and said species; and

D is H or alkyl, and n is an integer.

19. The composition of matter defined in claim 18, wherein G is selected from the group consisting of:

35. A compound having the structure

wherein Y is a linker arm.

36. A compound of claim 35, wherein Y has the structure

37. A compound of claim 36, wherein m is 3 and n is 2.

38. A composition of matter having the structure

39. A compound having the structure

wherein L is a bipyridyl or substituted bipyridyl bidentate ligand;

I is a diphosphine ligand; and

G is selected from the group consisting of;

40. The compound defined in claim 39, wherein I is:

41. A compound having the structure:

wherein G is selected from the group consisting of;

42. The compound of claim 41 wherein G reacts with a species selected from the group consisting of a nucleotide, amino acid, antibody, theophylline, digoxin, human chorionic gonadotropin, whole cell, subcellular particle, virus, prion, viroid, nucleic acid, protein, lipoprotein, lipopolysaccharide, glycoprotein, peptide, hormone, pharmacological agent, nonbiological polymer, synthetic organic molecule and an organometallic molecule, wherein G and said species react to form a covalent linkage between said compound and said species.

43. A compound having the structure:

wherein G is selected from the group consisting of

44. The compound of claim 43 wherein G reacts with a species selected from the group consisting of a nucleotide, amino acid, antibody, theophylline, digoxin, human chorionic gonadotropin, whole cell, subcellular particle, virus, prion, viroid, nucleic acid, protein, lipoprotein, lipopolysaccharide, glycoprotein, peptide, hormone, pharmacological agent, nonbiological polymer, synthetic organic molecule and an organometallic molecule, wherein G and said species react to form a covalent linkage between said compound and said species.

45. A compound having the structure:

wherein G is selected from the group consisting of;

46. A compound having the structure:

wherein G is selected from the group consisting of;