CA2421552A1 - Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses - Google Patents

Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses Download PDF

Info

Publication number
CA2421552A1
CA2421552A1 CA002421552A CA2421552A CA2421552A1 CA 2421552 A1 CA2421552 A1 CA 2421552A1 CA 002421552 A CA002421552 A CA 002421552A CA 2421552 A CA2421552 A CA 2421552A CA 2421552 A1 CA2421552 A1 CA 2421552A1
Authority
CA
Canada
Prior art keywords
group
dye
labeling reagent
target
labeled target
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CA002421552A
Other languages
French (fr)
Other versions
CA2421552C (en
Inventor
Jannis G. Stavrianopoulos
Elazar Rabbani
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Enzo Life Sciences Inc
Original Assignee
Enzo Life Sciences Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Enzo Life Sciences Inc filed Critical Enzo Life Sciences Inc
Priority to CA2753910A priority Critical patent/CA2753910C/en
Priority to CA2754041A priority patent/CA2754041C/en
Priority to CA2753922A priority patent/CA2753922C/en
Priority to CA2754186A priority patent/CA2754186C/en
Priority to CA2754415A priority patent/CA2754415C/en
Priority to CA2754185A priority patent/CA2754185C/en
Publication of CA2421552A1 publication Critical patent/CA2421552A1/en
Application granted granted Critical
Publication of CA2421552C publication Critical patent/CA2421552C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07HSUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09BORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B23/00Methine or polymethine dyes, e.g. cyanine dyes
    • C09B23/02Methine or polymethine dyes, e.g. cyanine dyes the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09BORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B47/00Porphines; Azaporphines
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09BORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B57/00Other synthetic dyes of known constitution
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10STECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00Chemistry: analytical and immunological testing
    • Y10S436/80Fluorescent 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 (286)

WHAT IS CLAIMED IS:
1. A labeling reagent for labeling a target, said labeling reagent comprising a marker moiety M and a reactive group R
M-R
wherein said marker moiety M and said reactive group R are covalently linked together, sand M comprising at least one moiety that comprises a ligand, a dye, or both a ligand and a dye; and said reactive group R being capable of forming a carbon-carbon linkage with said target.
2. The labeling reagent of claim 1, wherein said target comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.
3. The labeling reagent of claim 2, wherein said nucleic acid or nucleotide or nucleotide analog is modified.
4. The labeling reagent of claim 1, wherein said ligand, comprises biotin, iminobiotin, digoxygenin or fluorescein.
5. The labeling reagent of claim 1, wherein said dye comprises a fluorescent dye.
6. The labeling reagent of claim 5, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
7. The labeling reagent of claim 6, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
8. The labeling reagent of claim 1, further comprising a linker arm between said marker moiety M and said reactive group R.
9. The labeling reagent of claim 8, wherein said linker arm is flexible, semi-rigid or rigid.
10. The labeling reagent of claim 8, wherein said linker arm has a backbone comprising one or more carbon atoms.
11. The labeling reagent of claim 8, wherein said linker arm has a backbone comprising at least one non-carbon atom.
12. The labeling reagent of claim 11, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
13. The labeling reagent of claim 10, wherein said backbone comprises one or more peptide bonds.
14. The labeling reagent of claim 1, wherein said reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
15. The labeling reagent of claim 14, wherein said metallo-organic compound comprises mercury, zinc, copper or platinum.
16. The labeling reagent of claim 14, wherein said metallo-organic compound comprises an aromatic group, an aromatic heterocyclic group, an alkene group or an alkyne group.
17. A process for labeling a target, said process comprising the steps of:
(a) providing:
(i) sand target;
(ii) a Labeling reagent comprising a marker moiety M and a reactive group R
M - R
wherein said marker moiety M and said reactive group R are covalently linked together, said M comprising at least one moiety that comprises a ligand, a dye, or both a ligand and a dye; and said reactive group R being capable of forming a carbon-carbon linkage with said target; and (b) reacting said target (i) and said labeling reagent (ii) under conditions such that a carbon-carbon linkage forms between said target (;) and said labeling reagent (ii), thereby labeling said target (i) with said marker moiety M.
18. The process of claim 17, wherein in said providing step, the target comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.
19. The process of claim 17, wherein in said providing step, the ligand comprises biotin, iminobiotin, digoxygenin or fluorescein, and the dye comprises a fluorescent dye.
20. The process of claim 19, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
21. The process of claim 17, wherein said providing or reacting step, the reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
22. The process of claim 17, wherein in said providing step, the target has been modified.
23. The process of claim 22, wherein said modified target comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
24. A labeled target, said target having been labeled by a process comprising the steps of:
(a) providing:
(i) said target;
(ii) a labeling reagent comprising a marker moiety M and a reactive group R
M - R
wherein said marker moiety M and said reactive group R are covalently linked together, said M comprising at least one moiety that comprises a ligand, a dye, or both a ligand and a dye; and said reactive group R being capable of forming a carbon-carbon linkage with said target;
(b) reacting said target (i) and said labeling reagent (ii) under conditions such that a carbon-carbon linkage forms between said target (i) and said labeling reagent (ii), thereby labeling said target (i) with said marker moiety M.
25. The process of claim 24, wherein in said providing step, the target comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.
26. The process of claim 24, wherein in said providing step, the ligand comprises biotin, iminobiotin, digoxygenin or fluorescein, and the dye comprises a fluorescent dye.
27. The process of claim 26, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
28. The process of claim 24, wherein said providing ar reacting step, the reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metall o-organic compound.
29. The process of claim 24, wherein in said providing step, the target has been modified.
30. The process of claim 29, wherein said modified target comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
31. A process for preparing a cyanine dye labeling reagent, said process comprising the steps of:
(a) providing:
(i) a first intermediate compound comprising:
wherein X1 comprises carbon, oxygen, nitrogen or sulfur; and (ii) a second intermediate compound comprising:
wherein X1 comprises carbon, oxygen, nitrogen or sulfur;
wherein at least one of R1 through R10 comprises a reactive group capable of forming a carbon-carbon linkage with a target, and (ii) linking reagents suitable for linking said first intermediate compound and said second intermediate compound;
(b) forming a reaction mixture comprising said first intermediate compound (i), said second intermediate compound (ii), and said (inking reagents under conditions to link (i) and (ii) to form wherein at least one of R1 through R10 comprises a reactive group capable of forming a carbon-carbon linkage with a target, and wherein n is an integer of 1, 2 or 3, and wherein X1 and X2 independently comprise carbon, oxygen, nitrogen or sulfur.
32. The process of claim 31, wherein said providing step, the reactive group comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
33. The process of claim 31, wherein R1 through R10 independently comprise hydrogen, C1-C6 alkyl, a C0-C4 alkyl group having a hydrophilic substituent comprising sulfonate, carboxylate, hydroxyl, substituted amines and quaternary amines, aliphatic, alkenes, alkynes, charged or polar groups, or combinations of any of the foregoing.
34. A labeling reagent comprising an aphenylic analog of a rhodamine dye, said analog comprising at least one reactive group for attaching said labeling reagent to a target, said at least one reactive group being attached directly to said analog or indirectly through a linker arm.
35. The labeling reagent of claim 34, said reagent having the structure wherein R is a reactive group for attaching said labeling reagent to a target, and wherein R is attached directly or indirectly through an aphenylic linker arm.
36. The labeling reagent of claim 34, said reagent having the structure wherein R is a reactive group for attaching said labeling reagent to a target, and wherein R is attached directly or indirectly through an aphenylic linker arm.
37. The labeling reagent of claim 34, 35 or 36, wherein said reactive group comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.
38. The labeling reagent of claim 34, 35 or 36, wherein said reactive group is capable of forming a carbon-carbon linkage with a target.
39. The labeling reagent of claim 38, wherein said reactive group comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
40. The labeling reagent of claim 39, wherein said metallo-organic compound comprises mercury, zinc, copper or platinum.
41. The labeling reagent of claim 39, wherein said metallo-organic compound comprises an alkene group or an alkyne group.
42. The labeling reagent of claim 34, wherein said aphenylic linker arm has a backbone comprising one or more carbon atoms.
43. The labeling reagent of claim 34, wherein said aphenylic linker arm has a backbone comprising at least one non-carbon atom.
44. A labeled nucleotide comprising an aphenylic analog of a rhodamine dye, wherein said dye is attached directly to said nucleotide or indirectly through a linker.
45. The labeled nucleotide of claim 44, said labeled nucleotide having the structure:
wherein Nt comprises a nucleotide.
46. The labeled nucleotide of claim 44, said labeled nucleotide having the structure:
wherein Nt comprises a nucleotide.
47. The labeled nucleotide of claims 44, 45 or 46, wherein said nucleotide comprises a mono-phosphate, a di-phosphate or a tri-phosphate.
48. The labeled nucleotide of claims 44, 45 or 46, wherein said linker arm has a backbone comprising one or more carbon atoms.
49. The labeled nucleotide of claims 44, 45 or 46, wherein said linker arm has a backbone comprising at least one non-carbon atom.
50. The labeled nucleotide of claims 44, 45 or 46, wherein said linker arm comprises two or more consecutive peptide bonds.
51. A labeled target comprising T -- L -- M
wherein T is a target, M is a marker moiety and L is a chemical group covalently linking said M to T, said chemical group L comprising a backbone that comprises at least one rigid group that comprises one or more of:
(d) multimers of (a), (b) or (c), and (e) any combinations of (a), (b), (c) and (d).
52. The labeled target of claim 51, wherein said target T comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo-or polysaccharide.
53. The labeled target of claim 52, wherein said nucleic acid or nucleotide or nucleotide analog is modified.
54. The labeled target of claim 52, wherein said ligand comprises biotin, iminobiotin, digoxygenin or fluorescein.
55. The labeled target of claim 51, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxygenin or fluorescein.
56. The labeled target of claim 51, wherein said marker moiety M comprises a fluorescent dye.
57. The labeled target of claim 56, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
58. The labeled target of claim 57, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
59. The labeled target of claim 51, wherein said backbone comprises one or more carbon atoms.
60. The labeled target of claim 51, wherein said backbone comprises at least one non-carbon atom.
61. The labeled target of claim 60, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
62. The labeled target of claim 51, wherein said chemical group L further comprises at least one additional moiety comprising peptide bonds, amino acids, aliphatic chains from C1 through C20, alkene groups, alkyne groups, saturated or unsaturated or partially saturated rings, heterocyclic rings and sugars.
63. The labeled target of claim 51, wherein said structure (a) comprises two alkene groups in cis conformation to each other.
64. The labeled target of claim 51, wherein said structure (a) comprises two alkene groups in trans conformation to each other.
65. The labeling reagent of claim 51, wherein one or more hydrogens are substituted by a chemical moiety.
66. A labeling reagent comprising R -- L -- M
wherein R is a reactive group, M is a marker moiety and L is a chemical group covalently linking said M to R, said chemical group L comprising a backbone that comprises at least one rigid group that comprises one or more of:
(d) multimers of (a), (b) or (c), and (e) any combinations of (a), (b), (c) and (d).
67. The labeling reagent of claim 66, wherein said reactive group R comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isacyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or oldehyde.
68. The labeling reagent of claim 67, wherein as a reactive group R is capable of forming a carbon-carbon linkage with a target.
69. The labeling reagent of claim 68, wherein said reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
70. The labeling reagent of claim 69, wherein said metallo-organic compound comprises mercury, zinc, copper or platinum.
71. The labeling reagent of claim 69, wherein said metallo-organic compound comprises an aromatic group, an aromatic heterocyclic group, an alkene group or an alkyne group.
72. The labeling reagent of claim 66, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxygenin or fluorescein.
73. The labeling reagent of claim 66, wherein said marker moiety M comprises a fluorescent dye.
74. The labeling reagent of claim 73, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
75. The labeling reagent of claim 74, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
76. The labeling reagent of claim 66, wherein said backbone comprises one or more carbon atoms.
77. The labeling reagent of claim 66, wherein said backbone comprises at least one non-carbon atom.
78. The labeling reagent of claim 77, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
79. The labeling reagent of claim 77, wherein said chemical group L further comprises at least one additional moiety comprising peptide bonds, amino acids, aliphatic chains from C1 through C20, alkene groups, alkyne groups, saturated or unsaturated or partially saturated rings, heterocyclic rings and sugars.
80. The labeling reagent of claim 66, wherein said structure (a) comprises two alkene groups in cis conformation to each other.
81. The labeling reagent of claim 66, wherein said structure (a) comprises two alkene groups in trans conformation to each other.
82. The labeling reagent of claim 66, wherein one or more hydrogens are substituted by a chemical moiety.
83. A labeled target comprising T -- L -- M
wherein T is a target, M is a marker moiety and L is a chemical group covalently linking said M to T, said chemical group L comprising a backbone that comprises at least two consecutive polar rigid units.
84. The labeled target of claim 83, wherein said target T comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo-or polysaccharide.
85. The labeled target of claim 84, wherein said nucleic acid or nucleotide or nucleotide analog is modified.
86. The labeled target of claim 84, wherein said ligand comprises biotin, iminobiotin, digoxygenin or fluorescein.
87. The labeled target of claim 83, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxygenin or fluorescein.
88. The labeled target of claim 83, wherein said marker moiety M comprises a fluorescent dye.
89. The labeled target of claim 88, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
90. The labeled target of claim 89, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
91. The labeled target of claim 83, wherein at least one of said two consecutive rigid polar units of said backbone is heteroatomic.
92. The labeled target of claim 91, wherein said heteroatomic backbone comprises a carbon atom that is bonded to a N, S, O, P or halogen atom.
93. The labeled target of claim 91, wherein said heteroatom comprises an -OH, -SH, -SO3, -PO4, -COOH or -NH2 groups.
94. The labeled target of claim 91, wherein at least one of said two consecutive rigid polar units comprises a peptide bond.
95. The labeled target of claim 91, wherein at least one of said two consecutive polar units comprises a ring structure.
96. The labeled target of claim 95, wherein said ring structure further comprises polar or charged functional groups attached to said ring.
97. The labeled target of claim 96, wherein said polar or charged functional groups comprise halide, -OH, -SH, -SO3, -PO4, -COOH or -NH2 groups.
98. The labeled target of claim 95, wherein said ring structure comprises a sugar.
99. The labeled target of claim 95, wherein said ring structure comprises a substituted heterocyclic aromatic compound.
100. The labeled target of claim 83, wherein said backbone comprises one or more carbon atoms.
101. The labeled target of claim 83, wherein said backbone comprises at least one non-carbon atom.
102. The labeled target of claim 101, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
103. The labeled target of claim 83, wherein said chemical group L further comprises at least one additional moiety comprising peptide bonds, amino acids, aliphatic chains from C1 through C20, alkene groups, alkyne groups, saturated or unsaturated or partially saturated rings, heterocyclic rings and sugars.
104. A labeling reagent comprising R -- L -- M
wherein R is a reactive group, M is a marker moiety and L is a chemical group covalently linking said M to R, said chemical group L comprising a backbone that comprises at least two consecutive polar rigid units.
105. The labeling reagent of claim 104, wherein said reactive group R
comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.
106. The labeling reagent of claim 104, wherein as a reactive group R is capable of forming a carbon-carbon linkage with a target.
107. The labeling reagent of claim 104, wherein said reactive group R
comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
108. The labeling reagent of claim 107, wherein said metallo-organic compound comprises mercury, zinc, copper or platinum.
109. The labeling reagent of claim 107, wherein said metallo-organic compound comprises an aromatic group, an aromatic heterocyclic group, an alkene group or an alkyne group.
110. The labeling reagent of claim 104, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxygenin or fluorescein.
111. The labeling reagent of claim 104, wherein said marker moiety M comprises a fluorescent dye.
112. The labeling reagent of claim 111, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
113. The labeling reagent of claim 112, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
114. The labeled target of claim 104, wherein at least one of said two consecutive rigid polar units is heteroatomic.
115. The labeled target of claim 114, wherein said heteroatomic rigid polar units comprises a carbon atom that is bonded to a N, S, O, P or halogen atom.
116. The labeled target of claim 114, wherein said heteroatom comprises an -OH;
-SH, -SO3, -PO4, -COOH or -NH2 groups.
117. The labeled target of claim 104, wherein at least one of said two consecutive rigid polar units comprises a peptide bond.
118. The labeled target of claim 1049 wherein at least one of said two consecutive polar units comprises a ring structure.
119. The labeled target of claim 118, wherein said ring structure further comprises polar or charged functional groups attached to said ring.
120. The labeled target of claim 119, wherein said polar or charged functional groups comprise halide; -OH, -SH, -SO3, -PO4, -COOH or -NH2 groups.
121. The labeled target of claim 118, wherein said ring structure comprises a sugar.
122. The labeled target of claim 118, wherein said ring structure comprises a substituted heterocyclic aromatic compound.
123. The labeling reagent of claim 104, wherein said backbone comprises one or more carbon atoms.
124. The labeling reagent of claim 104, wherein said backbone comprises at least one non-carbon atom.
125. The labeling reagent of claim 124, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
126. The labeling reagent of claim 104, wherein said chemical group L further comprises at least one additional moiety comprising peptide bonds, amino acids, aliphatic chains from C1 through C20, alkene groups, alkyne groups, saturated or unsaturated or partially saturated rings, heterocyclic rings and sugars.
127. A labeled target comprising T -- L -- M
wherein T is a target, M is a marker moiety and L is a chemical group covalently linking said M to T, said chemical group L comprising a backbone that comprises at least two consecutive peptide bonds.
128. The labeled target of claim 242, wherein said target T comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.
129. The labeled target of claim 128, wherein said nucleic acid or nucleotide or nucleotide analog is modified.
130. The labeled target of claim 128, wherein said ligand comprises biotin, iminobiotin, digoxygenin or fluorescein.
131. The labeled target of claim 127, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxygenin or fluorescein.
132. The labeled target of claim 127, wherein said marker moiety M comprises a fluorescent dye.
133. The labeled target of claim 132, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
134. The labeled target of claim 133, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
135. The labeled target of claim 127, wherein at least one of said two consecutive peptide bonds are separated by a single atom.
136. The labeled target of claim 135, wherein said single; atom comprises C, N, S, O or P.
137. The labeled target of claim 127, wherein said backbone comprises one or more carbon atoms.
138. The labeled target of claim 127, wherein said backbone comprises at least one non-carbon atom.
139. The labeled target of claim 138, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
140. The labeled target of claim 127, wherein said chemical group L further comprises at least one additional moiety comprising peptide bonds, amino acids, aliphatic chains from C1 through C20, alkene groups, alkyne groups, saturated or unsaturated or partially saturated rings, heterocyclic rings and sugars.
141. The labeled target of claim 127, wherein said chemical group L comprises a di-peptide or an oligo-peptide.
142. The labeled target of claim 141, wherein said di-peptide or- oligo-peptide comprises (glycine)2 or (glycine)4.
143. A labeling reagent comprising R - L - M
wherein R is a reactive group, M is a marker moiety and L is a chemical group covalently linking said M to R, said chemical group L comprising a backbone that comprises at least two consecutive peptide bonds.
144. The labeling reagent of claim 143, wherein said reactive group R
comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.
145. The labeling reagent of claim 143, wherein as a reactive group R is capable of forming a carbon-carbon Linkage with a target.
146. The labeling reagent of claim 143, wherein said reactive group R
comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
147. The labeling reagent of claim 146, wherein said metallo-organic compound comprises mercury, zinc, copper or platinum.
148. The labeling reagent of claim 146, wherein said metallo-organic compound comprises an aromatic group, an aromatic heterocyclic group, an alkene group or an alkyne group.
149. The labeling reagent of claim 143, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxygenin or fluorescein.
150. The labeling reagent of claim 143, wherein said marker moiety M comprises a fluorescent dye.
151. The labeling reagent of claim 150, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.
152. The labeling reagent of claim 151, wherein said xanthene dye comprises fluorescein, rhodamine or rhodol, or derivatives thereof.
153. The labeled target of claim 143, wherein at least one of said two consecutive peptide bonds are separated by a single atom.
154. The labeled target of claim 153, wherein said single atom comprises C, N, S, O or P.
155. The labeled target of claim 143, wherein said backbone comprises one or more carbon atoms.
156. The labeled target of claim 143, wherein said backbone comprises at least one non-carbon atom.
157. The labeled target of claim 143, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.
158. The labeled target of claim 143, wherein said chemical group L further comprises at least one additional moiety comprising peptide bonds, amino acids, aliphatic chains from C1 through C20, alkene groups, alkyne groups, saturated or unsaturated or partially saturated rings, heterocyclic rings and sugars.
159. The labeled target of claim 143, wherein said chemical group L comprises a di-peptide or an oligo-peptide.
160. The labeled target of claim 159, wherein said di-peptide or oligo-peptide comprises (glycine)2 or (glycine)4.
167. A labeling reagent comprising a nonmetallic porphyrin, said reagent comprising:
wherein R0 is a reactive group and is attached directly or indirectly to said nonmetallic porphyrin, and R1 through R8 independently comprise hydrogen, aliphatic, unsaturated aliphatic, cyclic, heterocyclic, aromatic, heteroaromatic, charged or polar groups, or any combinations of the foregoing.
162. The labeling reagent of claim 161, wherein said reactive group R0 is attached directly or indirectly to any one of the four non-pyrrole positions.
163. The labeling reagent of claim 161, wherein said reactive group R0 comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.
164. The labeling reagent of claim 161, wherein as a reactive group R0 is capable of forming a carbon-carbon linkage with a target.
165. The labeling reagent of claim 161, wherein said reactive group R0 comprises an alkene group, an alkyne group or a halogenated compound.
166. The labeling reagent of claim 161, wherein any of said R1 through R8 alkyl groups comprises methyl, ethyl or propyl.
167. The labeling reagent of claim 161, wherein any of said R1 through R8 alkyl groups further comprises a polar or charged group.
168. The labeling reagent of claim 161, wherein said reactive group R0 is attached indirectly to said nonmetallic porphyrin through a linker arm.
169. The labeling reagent of claim 168, wherein said linker arm comprises at least two consecutive peptide bonds.
170. A labeled target comprising a nonmetallic porphyrin, said reagent comprising:
wherein T is a target molecule attached directly or indirectly to said nonmetallic porphyrin and R1 through R8 independently comprise hydrogen, aliphatic, unsaturated aliphatic, cyclic, heterocyclic, aromatic, heteroaromatic, charged or polar groups, or any combinations of the foregoing.
171. The labeled target of claim 170, wherein said reactive group R0 is attached directly or indirectly to any one of the tour non-pyrrole positions.
172. The labeled target of claim 170, wherein said target T comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a hapten, an antibody, a carbohydrate, a sugar or an oligo-or polysaccharide.
173. The labeled target of claim 172, wherein said nucleic acid or nucleotide or nucleotide analog is modified.
174. The labeling reagent of claim 170, wherein any of said R1 through R8 alkyl groups comprises methyl, ethyl or propyl.
175. The labeling reagent of claim 170, wherein any of said R1 through R8 alkyl groups further comprises a polar or charged group.
176. The labeling reagent of claim 170, wherein said target T is attached indirectly to said nonmetallic porphyrin through a linker arm.
177. The labeling reagent of claim 176, wherein said linker arm comprises at least two consecutive peptide bonds.
178. A process for determining the amount of a nucleic acid in a sample of interest, said process comprising the steps of:
(a) providing:
(i) said sample of interest;
(ii) a dye comprising a first phenanthridinium moiety linked to a second phenanthridinium moiety through a phenyl group in each of said first and second phenanthridinium moieties;
(iii) reagents for carrying out dye binding, hybridization, strand extension, or any combination thereof;
(b) forming a mixture of (i), (ii) and (iii) above, to produce a complex comprising said dye (ii) and any nucleic acid that may be present in said sample of interest (i);
(c) illuminating said mixture formed in step (b) at a wavelength below 400 nanometers (nm); and (d) measuring fluorescent emission from said illuminated mixture in step (c), said emission being proportional to the quantity of any nucleic acid present in said sample of interest (i).
179. The process of claim 178, wherein said sample of interest (i) is suspected of containing a nucleic acid or nucleic acids of interest.
180. The process of claim 179, wherein said nucleic acid or nucleic acids are double-stranded, single-stranded or a mixture of double-stranded and single-stranded nucleic acids
181. The process of claim 179, wherein said nucleic acid or nucleic acids comprise DNA, RNA or a mixture of RNA and DNA.
182. The process of claim 178, wherein said dye (ii) has the structure
183. The process of claim 179, further comprising the step (b') of hybridizing said nucleic acid or nucleic acids of interest to complementary nucleic acid strands or sequences fixed or immobilized to a solid support.
184. The process of 183, wherein said solid support comprises beads, tubes, microtiter plates, glass slides, plastic slides, microchip arrays, well or depressions.
185. The process of claim 178, wherein said illuminating step (c) is carried out at 350 manometers (nm).
186. The process of claim 178, wherein said measuring step (d) is carried out at greater than 550 manometers (nm).
187. The process of claim 186, wherein said measuring step (d) is carried out at 600 nanometers (nm).
188. The process of claim 178, wherein said measuring step (d) is carried out after or during a primer binding step, a primer extension step, a denaturation step or a step that is added for the purpose of fluoresence measurement
189. The process of claim 178, wherein said any of said process steps are carried out in a closed container.
190. The process of claim 189, wherein said closed container system comprises an illumination source and a detection device or unit.
191. A composition comprising at least one of the following dye structures:
192. The composition of claim 191, wherein the phenyl ring has been substituted in the ortho, para or meta positions.
193. The composition of claim 192, wherein said substitution is in the meta position of the phenyl ring.
194. A process for determining the amount of a nucleic acid in a sample of interest, said process comprising the steps of:
(a) providing:
(i) said sample of interest;
{ii) the dye (a), (b), (c) or (d) from the composition of claim 191;
(iii) reagents for carrying out dye binding, hybridization, strand extension, or any combination thereof;
(b) forming a mixture of (i), (ii) and (iii) above, to produce a complex comprising said dye (ii) and any nucleic acid that may be present in said sample of interest (i);
(c) illuminating said mixture formed in step (b) at a first wavelength; and (d) measuring at a second wavelength any fluorescent emission from said illuminated mixture in step (c), said emission being proportional to the quantity of any nucleic acid present in said sample of interest (i).
195. The process of claim 194, wherein said sample of interest (i) is suspected of containing a nucleic acid or nucleic acids of interest.
196. The process of claim 195, wherein said nucleic acid or nucleic acids are double-stranded, single-stranded or a mixture of double-stranded and single-stranded nucleic acids
197. The process of claim 195, wherein said nucleic acid or nucleic acids comprise DNA, RNA or a mixture of RNA and DNA.
198. The process of claim 195, further comprising the step (b') of hybridizing said nucleic acid or nucleic acids of interest to complementary nucleic acid strands or sequences fixed or immobilized to a solid support.
199. The process of 198, wherein said solid support comprises beads, tubes, microtiter plates, glass slides, plastic slides, microchip arrays, well or depressions.
200. The process of claim 194, wherein said measuring step (d) is carried out after or during a primer binding step, a primer extension step, a denaturation step or a step that is added for the purpose of fluoresence measurement
201. The process of claim 194, wherein said any of said process steps are carried out in a closed container.
202. The process of claim 201, wherein said closed container system comprises an illumination source and a detection device or unit.
203. A heterodimeric dye composition, said composition comprising a first dye that comprises a phenanthridinium moiety; and a second dye that is different from said first dye, said second dye being attached through the phenyl ring of said phenanthridium moiety.
204. The heterodimeric dye composition of claim 203, herring the structure wherein said second dye is not a phenanthridinium moiety.
205. The heterodimeric dye composition of claim 204, wherein said Second Dye is substituted in any of the ortho, mete or pare positions of the phenyl ring.
206. The heterodimeric dye composition of claim 205, wherein said Second Dye is substituted in the meta position of the phenyl ring.
207. The composition of claims 203 or 204, wherein said second dye comprises a thiazole, a fluorescein, a thiazole derivative, or a fluorescein derivative.
208. A process for determining the amount of a nucleic acid in a sample of interest, said process comprising the steps of:
(a) providing:
(i) said sample of interest;
(ii) the dye of claim 203;
(iii} reagents for carrying out dye binding, hybridization, strand extension, or any combination thereof;
(b) forming a mixture of (i), (ii) and (iii) above, to produce a complex comprising said dye (ii) and any nucleic acid that may be present in said sample of interest (i);
(c) illuminating said mixture formed in step (b) at a first wavelength; and (d) measuring at a second wavelength any fluorescent emission from said illuminated mixture in step (c), said emission being proportional to the quantity of any nucleic acid present in said sample of interest (i).
209. The process of claim 208, wherein said sample of interest (i) is suspected of containing a nucleic acid or nucleic acids of interest.
210. The process of claim 209, wherein said nucleic acid or nucleic acids are double-stranded, single-stranded or a mixture of double-stranded and single-stranded nucleic acids.
211. The process of claim 209, wherein said nucleic acid or nucleic acids comprise DNA, RNA or a mixture of RNA and DNA.
212. The process of claim 208, further comprising the step (b') of hybridizing said nucleic acid or nucleic acids of interest to complementary nucleic acid strands or sequences fixed or immobilized to a solid support.
213. The process of 212, wherein said solid support comprises beads, tubes, microtiter plates, glass slides, plastic slides, microchip arrays, well or depressions.
214. The process of claim 208, wherein said measuring step (d) is carried out after or during a primer binding step, a primer extension step, a denaturation step or a step that is added for the purpose of fluoresence measurement
215. The process of claim 208, wherein said any of said process steps are carried out in a closed container.
216. The process of claim 215, wherein said closed container system comprises an illumination source and a detection device or unit.
217. A chemiluminescent reagent having the structure:
wherein Q comprises a cycloalkyl or polycycloalkyl group attached covalently to the 4-membered ring portion of said dioxetane above directly or indirectly through a linkage group; wherein Z comprises hydrogen, alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl or cycloheteroalkyl; and wherein R1 and R2 comprise chemical moieties attached to different sites of a cyclic ring attached to said dioxetane, and wherein R1 is enzymatically converted into R1* which comprises a chemical reactive group G1, and wherein R2 is attached to said cyclic ring through an oxygen atom and comprises a chemical reactive group G2 which reacts with said G1 to convert said dioxetane to an unstable light-emitting dioxetane form.
218. The chemiluminescent reagent of claim 217, wherein said Q comprises an adamantyl group.
219. The chemiluminescent reagent of claim 217, wherein said cyclic ring comprises an aromatic ring.
220. The chemiluminescent reagent of claim 217, wherein R2 comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.
221. The chemiluminescent reagent of claim 220, wherein said substituted aliphatic group comprises halogen, nitrates, sulfonates or nitrites.
222. The chemiluminescent reagent of claim 217, wherein enzymatically convertible R1 comprises amides, esters, phosphates, carboxylic acids, fatty acids, glucose, xylose, fucose, or amino acids.
223. The chemiluminescent reagent of claim 217, wherein R1 is enzymatically converted into R1* through the action of enzymes comprising amidases, esterases, acetylcholinesterases, acid phosphatases, alkaline phosphatases, decarboxylases, lipases, glucosidases, xylosidases, fucosidases, trypsin or chymotrypsin.
224. The chemiluminescent reagent of claim 217, wherein after said enzymatic conversion of R1 to R1* and before said conversion of said dioxetane to the unstable light-emitting dioxetane form, an intermediate five- or six-membered ring is formed comprising a linkage between said G1 and G2.
225. 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) the chemiluminescent reagent of claim 217;
(ii) reagents and buffers for carrying out chemiluminescent reactions;
(b) forming a mixture of:
(1) (i), (ii) and (iii); or (2) (ii) and (iii) and contacting said mixture of (ii) and (iii) with (i);
(c) enzymatically converting the chemiluminescent reagent of claim 217 (ii) into an unstable light-emitting dioxetane form; and (d) measuring the quantity of light generated by said enzymatic conversion in step (c).
226. The process of claim 225, wherein in said providing step (a) Q in said chemiluminescent reagent (ii) comprises an adamantyl group.
227. The process of claim 225, wherein in said providing step (a) R2 in said chemiluminescent reagent (ii) comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.
228. The process of claim 227, wherein said substituted aliphatic group comprises halogen, nitrates, sulfonates or nitrites.
229. The process of claim 225, wherein said enzymatic converting step (c) is carried out by a substrate comprising amides, esters, phosphates, carboxylic acids, fatty acids, glucose, xylose, fucose, or amino acids.
230. The process of claim 225, wherein said enzymatic activity of interest comprises an amidase, an esterase, an acetylcholinesterase, an acid phosphatase, an alkaline phosphatase, a decarboxylase, a lipase, a glucosidase, a xylosidase, a fucosidase, a trypsin or a chymotrypsin.
231. The process of claim 225, 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).
232. The process of claim 225, wherein any of said steps (a) through (d) are carried out in liquid phase or mixed phase.
233. The process of claim 225, wherein said enzymatic activity of interest is dependent upon the presence or quantity of another compound.
234. The process of claim 233, wherein said another compound comprises an RNA
or DNA probe.
235. A chemiluminescent reagent having the structure:
wherein Q comprises a cycloalkyl or polycycloalkyl group attached covalently to the 4-membered ring portion of said dioxetane above directly or indirectly through a linkage group; wherein Z comprises hydrogen, alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl or cycloheteroalkyl and wherein R comprises a chemical linker having a reactive site attached to the aromatic ring in said structure; and wherein R' comprises a substrate for an non-cleaving enzymatic process, wherein the product of said enzymatic process leads to further chemical rearrangements that generate an unstable light emitting dioxetane form.
236. The chemiluminescent reagent of claim 235, wherein said non-cleaving enzymatic process comprises oxidation or reduction.
237. The chemiluminescent reagent of claim 235, wherein said Q comprises an adamantyl group.
238. The chemiluminescent reagent of claim 235, wherein R comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.
239. The chemiluminescent reagent of claim 238, wherein said substituted aliphatic group comprises halogen, nitrate, sulfonate or nitrite.
240. The chemiluminescent reagent of claim 235, wherein said reactive site comprises an oxygen, a nitrogen or a sulfur atom.
241. The chemiluminescent reagent of claim 235, wherein said enzymatic process is carried out by an enzyme comprising an oxidase or reductase.
242. The chemiluminescent reagent of claim 235, wherein after said enzymatic process, said dioxetane is converted to an unstable light-emitting dioxetane form.
243. 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) the chemiluminescent reagent of claim 235;
(ii) reagents and buffers for carrying out chemiluminescent reactions;
(b) forming a mixture of:
(1) (i), (ii) and (iii); or (2) (ii) and (iii) and contacting said mixture of (ii) and (iii) with (i);
(c) enzymatically converting the chemiluminescent reagent of claim 344 (ii) into an unstable light-emitting dioxetane form; and (d) measuring the quantity of light generated by said enzymatic conversion in step (c).
244. The process of claim 243, wherein in said providing step (a) Q in said chemiluminescent reagent (ii) comprises an adamantyl group.
245. The process of claim 243, wherein in said providing step (a) R in the chemiluminescent reagent (ii) comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.
246. The process of claim 245, wherein said substituted aliphatic group comprises halogen, nitrate, sulfonate or nitrite.
247. The process of claim 243, wherein said providing step (a) R in the chemiluminescent reagent (ii) comprises a reactive site comprising an oxygen, a nitrogen or a sulfur atom.
248. The process of claim 243, wherein said step of enzymatically converting (c) is carried out by an enzyme comprising an oxidase or reductase.
249. The process of claim 243, wherein any of said steps (a) through (d) are carried out in liquid phase or mixed phase.
250. The process of claim 243, wherein said enzymatic activity of interest is dependent upon the presence or quantity of another compound.
251. The process of claim 250, wherein said another compound comprises an RNA
or DNA probe.
252. A dye composition of the formula R - Fluorescent Dye wherein R is covalently linked to said Fluorescent Dye comprises two or more members in combination from a) unsaturated aliphatic groups;
b) unsaturated heterocyclic groups;
c) aromatic groups;
and wherein R is capable of providing a conjugated system or an electron delocalized system with said fluorescent dye.
253. The dye composition of claim 252, wherein said unsaturated aliphatic groups comprise an alkene or an alkyne.
254. The dye composition of claim 252, wherein said aromatic groups comprise a phenyl group, an aryl group or an aromatic heterocyclic group.
255. The dye composition of claims 253 or 254, wherein said unsaturated aliphatic groups or aromatic groups are substituted.
256. The dye composition of claim 255, wherein said substituted unsaturated aliphatic groups or substituted aromatic groups comprise alkyl groups, aryl groups, alkoxy groups, phenoxy groups, amines, amino groups, amido groups, carboxy groups, nitrates, nitrites, sulfonates, sulfhydryl groups or phosphates.
257. The dye composition of claim 255, wherein said substituted aromatic groups comprise a fused ring structure.
258. The dye composition of claim 257, wherein said fused ring structure is a naphthalene, anthracene or a phenanthrene.
259. The dye composition of claim 252, wherein said combination comprises two members of the same group or of different groups.
260. The dye composition of claim 259, wherein said different groups comprise an unsaturated aliphatic group (a) and an unsaturated heterocyclic group (b);
an unsaturated aliphatic group (a) and an aromatic group (c); or an unsaturated heterocyclic group (b) and an aromatic group (c).
261. The dye composition of claim 252, wherein said fluorescent dye comprises an anthracene, a xanthene, a cyanine, a porphyrin, a coumarin or a composite dye.
262. The dye composition of claim 252, further comprising a charged or polar R' group.
263. The dye composition of claim 262, wherein said charged or polar R' group increases aqueous solubility of said composition.
264. The dye composition of claim 253 or 262, further comprising a reactive group R x attached to either said fluorescent dye, said R group or said R' group.
265. The dye composition of claim 264, further comprising a linker arm attaching said reactive group R x to said fluorescent dye, said R group or said R' group.
266. The dye composition of claim 264, wherein said reactive group R x comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.
267. The dye composition of claim 265, wherein said reactive group R x comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.
268. The dye composition of claim 264, wherein as a reactive group R is capable of forming a carbon-carbon linkage with a target.
269. The dye composition of claim 265, wherein as a reactive group R is capable of forming a carbon-carbon linkage with a target.
270. The dye composition of claim 268, wherein said reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
271. The dye composition of claim 269, wherein said reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.
272. A labeled target having the structure wherein said Dye is a fluorescent dye, wherein R is covalently linked to said Dye, and wherein R comprises two or more members in combination from a) unsaturated aliphatic groups;
b) unsaturated heterocyclic groups;
c) aromatic groups;
and wherein R is capable of providing a conjugated system or an electron delocalized system with said Dye.
comprises two or more unsaturated aliphatic groups, unsaturated heterocyclic groups, aromatic groups, or combinations of the foregoing groups and wherein R is covalently attached to said fluorescent dye and is capable of providing a conjugated system or an electron delocalized system with said fluorescent dye, and wherein said target is covalently attached to said Dye or said R.
273. The labeled target of claim 272, wherein said unsaturated aliphatic groups comprise an alkene or an alkyne.
274. The labeled target of claim 272, wherein said aromatic groups comprise a phenyl group, an aryl group or an aromatic heterocyclic group.
275. The labeled target of claims 273 or 274, wherein said unsaturated aliphatic groups or aromatic groups are substituted.
276. The labeled target of claim 272, wherein said substituted unsaturated aliphatic groups or substituted aromatic groups comprise alkyl groups, aryl groups, alkoxy groups, phenoxy groups, amines, amino groups, amido groups, carboxy groups, nitrates, nitrites, sulfonates, sulfhydryl groups or phosphates.
277. The labeled target of claim 275, wherein said substituted aromatic groups comprise a fused ring structure.
278. The labeled target of claim 277, wherein said fused ring structure is a naphthalene, anthracene or a phenanthrene.
279. The labeled target of claim 272, wherein said combination comprises two members of the same group or of different groups.
280. The labeled target of claim 279, wherein said different groups comprise an unsaturated aliphatic group (a) and an unsaturated heterocyclic group (b);
an unsaturated aliphatic group (a) and an aromatic group (c); or an unsaturated heterocyclic group (b) and an aromatic group (c).
281. The labeled target of claim 272, wherein said fluorescent dye comprises an anthracene, a xanthene, a cyanine, a porphyrin, a coumarin or a composite dye.
282. The labeled target of claim 272, further comprising a charged or polar R' group
283. The labeled target of claim 272, wherein said charged or polar R' group increases aqueous solubility of said composition.
284. The labeled target of claim 282, further comprising a linker arm attaching said target to said fluorescent dye, said R group or said R' group.
285. The labeled target of claim 272, wherein said target comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.
286. The labeled target of claim 285, wherein said ligand comprises biotin, iminobiotin, digoxygenin or fluorescein, and the dye comprises a fluorescent dye.
CA2421552A 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses Expired - Lifetime CA2421552C (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
CA2753910A CA2753910C (en) 2002-03-12 2003-03-11 Fluorescent dye composition and target labeled therewith
CA2754041A CA2754041C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2753922A CA2753922C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754186A CA2754186C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754415A CA2754415C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754185A CA2754185C (en) 2002-03-12 2003-03-11 Chemiluminescent dioxetanes

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US10/096,075 US7166478B2 (en) 2002-03-12 2002-03-12 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
US10/096,075 2002-03-12

Related Child Applications (6)

Application Number Title Priority Date Filing Date
CA2754186A Division CA2754186C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2753922A Division CA2753922C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754041A Division CA2754041C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754415A Division CA2754415C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2753910A Division CA2753910C (en) 2002-03-12 2003-03-11 Fluorescent dye composition and target labeled therewith
CA2754185A Division CA2754185C (en) 2002-03-12 2003-03-11 Chemiluminescent dioxetanes

Publications (2)

Publication Number Publication Date
CA2421552A1 true CA2421552A1 (en) 2003-09-12
CA2421552C CA2421552C (en) 2011-11-22

Family

ID=27804274

Family Applications (7)

Application Number Title Priority Date Filing Date
CA2753910A Expired - Lifetime CA2753910C (en) 2002-03-12 2003-03-11 Fluorescent dye composition and target labeled therewith
CA2754415A Expired - Lifetime CA2754415C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2753922A Expired - Lifetime CA2753922C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2421552A Expired - Lifetime CA2421552C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754041A Expired - Lifetime CA2754041C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754186A Expired - Lifetime CA2754186C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754185A Expired - Lifetime CA2754185C (en) 2002-03-12 2003-03-11 Chemiluminescent dioxetanes

Family Applications Before (3)

Application Number Title Priority Date Filing Date
CA2753910A Expired - Lifetime CA2753910C (en) 2002-03-12 2003-03-11 Fluorescent dye composition and target labeled therewith
CA2754415A Expired - Lifetime CA2754415C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2753922A Expired - Lifetime CA2753922C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses

Family Applications After (3)

Application Number Title Priority Date Filing Date
CA2754041A Expired - Lifetime CA2754041C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754186A Expired - Lifetime CA2754186C (en) 2002-03-12 2003-03-11 Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
CA2754185A Expired - Lifetime CA2754185C (en) 2002-03-12 2003-03-11 Chemiluminescent dioxetanes

Country Status (5)

Country Link
US (11) US7166478B2 (en)
EP (1) EP1348713B1 (en)
JP (3) JP4551063B2 (en)
CA (7) CA2753910C (en)
IL (13) IL195306A0 (en)

Families Citing this family (50)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9261460B2 (en) * 2002-03-12 2016-02-16 Enzo Life Sciences, Inc. Real-time nucleic acid detection processes and compositions
AU2002324468A1 (en) * 2001-08-17 2003-03-03 Eli Lilly And Company Rapid improvement of cognition in conditions related to abeta
US7166478B2 (en) * 2002-03-12 2007-01-23 Enzo Life Sciences, Inc., C/O Enzo Biochem, Inc. Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
US9353405B2 (en) 2002-03-12 2016-05-31 Enzo Life Sciences, Inc. Optimized real time nucleic acid detection processes
US20050129691A1 (en) * 2002-04-25 2005-06-16 Eli Lily And Company Patent Division Method for treating anxiety and mood disorders in older subjects
US7575747B2 (en) 2003-02-10 2009-08-18 Applied Molecular Evolution Aβ binding molecules
EP1594987B1 (en) * 2003-02-21 2009-05-27 GeneForm Technologies Limited Nucleic acid sequencing methods, kits and reagents
US9696298B2 (en) 2003-04-03 2017-07-04 Enzo Life Sciences, Inc. Multisignal reagents for labeling analytes
US7514551B2 (en) * 2003-04-03 2009-04-07 Enzo Life Sciences, Inc. Multisignal labeling reagents, and processes and uses therefor
US9156986B2 (en) 2003-04-03 2015-10-13 Enzo Life Sciences, Inc. Multisignal labeling reagents and processes and uses therefor
US20050170367A1 (en) * 2003-06-10 2005-08-04 Quake Stephen R. Fluorescently labeled nucleoside triphosphates and analogs thereof for sequencing nucleic acids
ITPZ20030002A1 (en) * 2003-08-12 2005-02-13 Giuseppe Caputo COMPOUNDS OF THE CIANINA TYPE WITH AN ALCHINYLIC ARM
GB0324456D0 (en) * 2003-10-20 2003-11-19 Isis Innovation Parallel DNA sequencing methods
US8124334B2 (en) * 2004-07-06 2012-02-28 Enzo Biochem, Inc. Selective detection of oncogenic HPV
US7776567B2 (en) 2005-03-17 2010-08-17 Biotium, Inc. Dimeric and trimeric nucleic acid dyes, and associated systems and methods
US7601498B2 (en) * 2005-03-17 2009-10-13 Biotium, Inc. Methods of using dyes in association with nucleic acid staining or detection and associated technology
US8362250B2 (en) 2005-05-24 2013-01-29 Enzo Biochem, Inc. Fluorescent dyes and compounds, methods and kits useful for identifying specific organelles and regions in cells of interest
US8357801B2 (en) 2005-05-24 2013-01-22 Enzo Life Sciences, Inc. Labeling of target molecules, identification of organelles and other applications, novel compositions, methods and kits
US7737281B2 (en) * 2005-05-24 2010-06-15 Enzo Life Sciences, Inc. C/O Enzo Biochem, Inc. Purine based fluorescent dyes
US7569695B2 (en) * 2005-05-24 2009-08-04 Enzo Life Sciences, Inc. Dyes for the detection or quantification of desirable target molecules
US20070141597A1 (en) * 2005-10-25 2007-06-21 Harmon H J Biomimetic Biodetector of Toxins, Viruses, Bacteria, and Biological Factors
US7781187B2 (en) * 2005-12-30 2010-08-24 Corning Incorporated Fluorescent dyes
JP2007215496A (en) * 2006-02-17 2007-08-30 Filgen Inc Fluorescent labeling method of probe nucleic acid in nucleic acid analysis method
US20080091005A1 (en) * 2006-07-20 2008-04-17 Visigen Biotechnologies, Inc. Modified nucleotides, methods for making and using same
US7629125B2 (en) * 2006-11-16 2009-12-08 General Electric Company Sequential analysis of biological samples
SI2121754T1 (en) * 2007-01-18 2015-03-31 Eli Lilly And Company Pegylated amyloid beta fab
US7910754B2 (en) 2007-02-06 2011-03-22 Xerox Corporation Colorant compounds
US7485737B2 (en) * 2007-02-06 2009-02-03 Xerox Corporation Colorant compounds
US20090162845A1 (en) 2007-12-20 2009-06-25 Elazar Rabbani Affinity tag nucleic acid and protein compositions, and processes for using same
US9250249B2 (en) 2008-09-08 2016-02-02 Enzo Biochem, Inc. Autophagy and phospholipidosis pathway assays
US9334281B2 (en) * 2008-09-08 2016-05-10 Enzo Life Sciences, Inc. Fluorochromes for organelle tracing and multi-color imaging
US8658665B2 (en) * 2008-12-10 2014-02-25 Wista Laboratories Ltd. 3,6-disubstituted xanthylium salts and use thereof in treatment of tauopathies
US9133343B2 (en) 2009-11-30 2015-09-15 Enzo Biochem, Inc. Dyes and compositions, and processes for using same in analysis of protein aggregation and other applications
US8877437B1 (en) 2009-12-23 2014-11-04 Biotium, Inc. Methods of using dyes in association with nucleic acid staining or detection
US9493824B2 (en) 2010-12-16 2016-11-15 Bio-Rad Laboratories, Inc. Universal reference dye for quantitative amplification
EP2686385B1 (en) 2011-03-15 2019-01-16 Ramot at Tel Aviv University, Ltd. Activatable fluorogenic compounds and uses thereof as near infrared probes
US20120316319A1 (en) * 2011-06-10 2012-12-13 Alexey Ustinov Arylethynylxanthene dyes
WO2013039883A1 (en) 2011-09-12 2013-03-21 Abbvie Biotherapeutics Inc. Artificial nk cells and uses thereof
US9416153B2 (en) 2011-10-11 2016-08-16 Enzo Life Sciences, Inc. Fluorescent dyes
US10060910B2 (en) 2012-07-24 2018-08-28 Enzo Life Sciences, Inc. Multisignal reagents for labeling analytes
WO2014027652A1 (en) 2012-08-17 2014-02-20 独立行政法人科学技術振興機構 Method and device for biomolecule analysis using raman spectroscopy
EP2976622B1 (en) * 2013-03-14 2019-11-27 Bio-Rad Laboratories, Inc. Protein standard
US9957393B2 (en) 2015-03-30 2018-05-01 Enzo Biochem, Inc. Monoazo dyes with cyclic amine as fluorescence quenchers
EP3235909A1 (en) * 2016-04-20 2017-10-25 SFC Co., Ltd. Merocyanine-based compounds, and dyes, kits and contrast medium compositions for labelling biomolecules comprising the same
EP3464477A1 (en) * 2016-06-06 2019-04-10 Sony Corporation Ionic polymers comprising fluorescent or colored reporter groups
CN110426442B (en) * 2016-06-13 2022-02-11 苏州优逸兰迪生物科技有限公司 Method for testing nucleic acid gel electrophoresis
CN108047135B (en) * 2017-12-29 2019-12-17 苏州宇恒生物科技有限公司 fluorescent dye, preparation method thereof and application thereof in bacterial detection
US11384376B2 (en) 2018-05-31 2022-07-12 Roche Molecular Systems, Inc. Reagents and methods for post-synthetic modification of nucleic acids
WO2021123074A1 (en) 2019-12-18 2021-06-24 F. Hoffmann-La Roche Ag Methods of sequencing by synthesis using a consecutive labeling scheme
CN116926178A (en) * 2022-04-02 2023-10-24 深圳市真迈生物科技有限公司 Fluorescence detection reagent and preparation method and application thereof

Family Cites Families (93)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4375972A (en) 1979-10-17 1983-03-08 Allied Corporation Heterogeneous chemiluminescent immunoassays utilizing metallo porphyrin tag
US4711955A (en) * 1981-04-17 1987-12-08 Yale University Modified nucleotides and methods of preparing and using same
CA1190838A (en) 1981-07-17 1985-07-23 Cavit Akin Homogeneous nucleic acid hybridization diagnostics by non-radiative energy transfer
CA1186621A (en) * 1981-08-10 1985-05-07 John L. Hendrix Fluoro immuno assay system
US4707454A (en) 1981-08-10 1987-11-17 Bio-Diagnostics, Inc. Fluorescent chlorophyll labeled assay reagents
US5241060A (en) * 1982-06-23 1993-08-31 Enzo Diagnostics, Inc. Base moiety-labeled detectable nucleatide
IL70765A (en) * 1983-01-27 1988-07-31 Enzo Biochem Inc Substrates containing non-radioactive chemically-labeled polynucleotides and methods using them
US4994373A (en) 1983-01-27 1991-02-19 Enzo Biochem, Inc. Method and structures employing chemically-labelled polynucleotide probes
US5013831A (en) 1984-01-30 1991-05-07 Enzo Biochem, Inc. Detectable molecules, method of preparation and use
US4952685A (en) 1984-01-30 1990-08-28 Enzo Biochem, Inc. Detectable molecules, method of preparation and use
US5288609A (en) 1984-04-27 1994-02-22 Enzo Diagnostics, Inc. Capture sandwich hybridization method and composition
CA1260372A (en) 1984-04-27 1989-09-26 Elazar Rabbani Hybridization method for the detection of genetic materials
US6221581B1 (en) 1984-04-27 2001-04-24 Enzo Diagnostics, Inc. Processes for detecting polynucleotides, determining genetic mutations or defects in genetic material, separating or isolating nucleic acid of interest from samples, and useful compositions of matter and multihybrid complex compositions
ATE40572T1 (en) * 1985-01-10 1989-02-15 Molecular Diagnostics Inc PHOTOCHEMICAL METHOD FOR LABELING NUCLEIC ACIDS FOR DETECTION IN HYBRIDISATION SAMPLES.
US5104637A (en) * 1985-02-06 1992-04-14 University Of Cincinnati Radio labeled dihematophorphyrin ether and its use in detecting and treating neoplastic tissue
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4824659A (en) * 1985-06-07 1989-04-25 Immunomedics, Inc. Antibody conjugates
US4868103A (en) 1986-02-19 1989-09-19 Enzo Biochem, Inc. Analyte detection by means of energy transfer
US5627027A (en) 1986-04-18 1997-05-06 Carnegie Mellon University Cyanine dyes as labeling reagents for detection of biological and other materials by luminescence methods
US5268486A (en) 1986-04-18 1993-12-07 Carnegie-Mellon Unversity Method for labeling and detecting materials employing arylsulfonate cyanine dyes
US6225050B1 (en) * 1986-04-18 2001-05-01 Carnegie Mellon University Cyanine dyes as labeling reagents for detection of biological and other materials by luminescence methods
CA1340806C (en) * 1986-07-02 1999-11-02 James Merrill Prober Method, system and reagents for dna sequencing
US5047519A (en) 1986-07-02 1991-09-10 E. I. Du Pont De Nemours And Company Alkynylamino-nucleotides
US5707559A (en) 1986-07-17 1998-01-13 Tropix, Inc. Chemiluminescent 1,2-dioxetane compounds
US4978614A (en) 1988-10-26 1990-12-18 Tropix, Inc. Method of detecting a substance using enzymatically-induced decomposition of dioxetanes
US4952686A (en) * 1987-12-01 1990-08-28 Fmc Corporation Soluble dried cassia alloy gum composition and process for making same
US5130238A (en) 1988-06-24 1992-07-14 Cangene Corporation Enhanced nucleic acid amplification process
JPH0257978A (en) * 1988-08-24 1990-02-27 Takara Shuzo Co Ltd Method for determining arrangement of dna base
US5118801A (en) 1988-09-30 1992-06-02 The Public Health Research Institute Nucleic acid process containing improved molecular switch
US5132204A (en) 1989-05-31 1992-07-21 Chiron Corporation Chemiluminescent double-triggered 1, 2-dioxetanes
US5545522A (en) 1989-09-22 1996-08-13 Van Gelder; Russell N. Process for amplifying a target polynucleotide sequence using a single primer-promoter complex
FR2656866B1 (en) * 1990-01-10 1992-05-15 Cis Bio Int PORPHYRIN DERIVATIVES AND METALLOPORPHYRINS POTENTIALLY COUPLED TO A BIOLOGICALLY ACTIVE MOLECULE, AND PHARMACEUTICAL COMPOSITION CONTAINING THEM.
US5401847A (en) 1990-03-14 1995-03-28 Regents Of The University Of California DNA complexes with dyes designed for energy transfer as fluorescent markers
US5494810A (en) 1990-05-03 1996-02-27 Cornell Research Foundation, Inc. Thermostable ligase-mediated DNA amplifications system for the detection of genetic disease
US5455175A (en) 1990-06-04 1995-10-03 University Of Utah Research Foundation Rapid thermal cycling device
US5210015A (en) 1990-08-06 1993-05-11 Hoffman-La Roche Inc. Homogeneous assay system using the nuclease activity of a nucleic acid polymerase
US5455166A (en) 1991-01-31 1995-10-03 Becton, Dickinson And Company Strand displacement amplification
US5994056A (en) 1991-05-02 1999-11-30 Roche Molecular Systems, Inc. Homogeneous methods for nucleic acid amplification and detection
US5213831A (en) * 1991-05-06 1993-05-25 Brown's Best Food Processing system and method for quick-cooking legumes
US5248618A (en) 1991-06-05 1993-09-28 Life Technologies, Inc. Methods for generating light with chemiluminescent dioxetanes activated by anchimeric assisted cleavage
IL102523A0 (en) * 1991-07-31 1993-01-14 Du Pont Enhancing the chemiluminescence of enzyme-triggered 1,2-dioxetanes
JP2914405B2 (en) * 1991-08-07 1999-06-28 日石三菱株式会社 Method for producing labeled porphyrin compound
ATE189831T1 (en) 1991-11-14 2000-03-15 Dgi Inc NON-RADIOACTIVE HYBRIDIZATION TEST METHOD AND KIT
US5270184A (en) 1991-11-19 1993-12-14 Becton, Dickinson And Company Nucleic acid target generation
US5565552A (en) * 1992-01-21 1996-10-15 Pharmacyclics, Inc. Method of expanded porphyrin-oligonucleotide conjugate synthesis
US5393469A (en) * 1992-03-20 1995-02-28 Lumigen, Inc. Polymeric phosphonium salts providing enhanced chemiluminescence from 1,2-dioxetanes
KR100249110B1 (en) 1992-05-06 2000-04-01 다니엘 엘. 캐시앙 Nucleic acid sequence amplification method, composition and kit
US5340714A (en) * 1992-05-08 1994-08-23 Monitor Diagnostics, Inc. Use of nonmetallic tetrapyrrole molecules and novel signal solutions in chemiluminescent reactions and assays
US5462854A (en) 1993-04-19 1995-10-31 Beckman Instruments, Inc. Inverse linkage oligonucleotides for chemical and enzymatic processes
FR2706618B1 (en) 1993-06-11 1995-09-01 Bio Merieux Device for the dosage of haptens and its use.
US5428149A (en) * 1993-06-14 1995-06-27 Washington State University Research Foundation Method for palladium catalyzed carbon-carbon coulping and products
US5808077A (en) 1993-06-30 1998-09-15 Abbott Laboratories Intercalators having affinity for DNA and methods of use
DE69435240D1 (en) 1993-06-30 2009-10-29 Abbott Lab Intercalators with DNA affinity and application methods
JPH0794807A (en) * 1993-07-27 1995-04-07 Toshiba Corp Amorphous organic thin film element, amorphous organic polymer compound and amorphous inorganic compound
US5464741A (en) 1993-10-08 1995-11-07 Henwell, Inc. Palladium (II) octaethylporphine alpha-isothiocyanate as a phosphorescent label for immunoassays
CA2140081C (en) 1994-01-13 2008-04-01 Dean L. Engelhardt Process, construct and conjugate for producing multiple nucleic acid copies
US5578832A (en) 1994-09-02 1996-11-26 Affymetrix, Inc. Method and apparatus for imaging a sample on a device
DE4407423A1 (en) * 1994-03-05 1995-09-07 Boehringer Mannheim Gmbh Anti-interference agent for use in immunoassays
US6589736B1 (en) * 1994-11-22 2003-07-08 The Trustees Of Boston University Photocleavable agents and conjugates for the detection and isolation of biomolecules
US6008373A (en) 1995-06-07 1999-12-28 Carnegie Mellon University Fluorescent labeling complexes with large stokes shift formed by coupling together cyanine and other fluorochromes capable of resonance energy transfer
US5827073A (en) * 1995-07-05 1998-10-27 Ludwig Institute For Cancer Research Photoreactive peptide derivatives
US5929227A (en) * 1995-07-12 1999-07-27 The Regents Of The University Of California Dimeric fluorescent energy transfer dyes comprising asymmetric cyanine azole-indolenine chromophores
US5631167A (en) * 1995-07-31 1997-05-20 Bayer Corporation Capsule chemistry analytical methods employing dioxetane chemiluminescence
JP3439221B2 (en) 1995-12-18 2003-08-25 ワシントン ユニヴァーシティ Nucleic acid analysis method using fluorescence resonance energy transfer
GB9605690D0 (en) 1996-03-19 1996-05-22 Oxford Biosciences Ltd Particle delivery
US5945526A (en) 1996-05-03 1999-08-31 Perkin-Elmer Corporation Energy transfer dyes with enhanced fluorescence
US5800996A (en) * 1996-05-03 1998-09-01 The Perkin Elmer Corporation Energy transfer dyes with enchanced fluorescence
US6174670B1 (en) 1996-06-04 2001-01-16 University Of Utah Research Foundation Monitoring amplification of DNA during PCR
US6001573A (en) 1996-06-14 1999-12-14 Packard Bioscience B.V. Use of porphyrins as a universal label
US5866336A (en) 1996-07-16 1999-02-02 Oncor, Inc. Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon
US5730849A (en) 1996-09-30 1998-03-24 Bio-Rad Laboratories, Inc. High sensitivity dyes as stains for RNA bands in denaturing gels
ES2182121T3 (en) * 1996-10-08 2003-03-01 Kreatech Biotech Bv METHODS TO MARK NUCLEOTIDES, MARKED NUCLEOTIDES AND USEFUL INTERMEDIARIES.
US5800999A (en) * 1996-12-16 1998-09-01 Tropix, Inc. Dioxetane-precursor-labeled probes and detection assays employing the same
US6506895B2 (en) * 1997-08-15 2003-01-14 Surmodics, Inc. Photoactivatable nucleic acids
US6008379A (en) * 1997-10-01 1999-12-28 The Perkin-Elmer Corporation Aromatic-substituted xanthene dyes
JP3595137B2 (en) 1997-11-14 2004-12-02 シャープ株式会社 LCD tablet device
GB9725197D0 (en) 1997-11-29 1998-01-28 Secr Defence Detection system
US6080868A (en) * 1998-01-23 2000-06-27 The Perkin-Elmer Corporation Nitro-substituted non-fluorescent asymmetric cyanine dye compounds
AU748847B2 (en) 1998-02-27 2002-06-13 Pamgene B.V. Method for the non-specific amplification of nucleic acid
US6068979A (en) * 1998-04-22 2000-05-30 Lumigen, Inc. Simplified sequential chemiluminescent detection
US5948648A (en) 1998-05-29 1999-09-07 Khan; Shaheer H. Nucleotide compounds including a rigid linker
US6110630A (en) 1998-06-18 2000-08-29 Beckman Coulter, Inc. Efficient activated cyanine dyes
JP4519320B2 (en) * 1998-07-28 2010-08-04 アプライド バイオシステムズ, エルエルシー Benzothiazole dioxetanes
US6288221B1 (en) * 1998-08-20 2001-09-11 Duke University Methods of synthesis of halogen base-modified oligonucleotides and subsequent labeling with a metal-catalyzed reaction
US6114350A (en) 1999-04-19 2000-09-05 Nen Life Science Products, Inc. Cyanine dyes and synthesis methods thereof
CA2376955A1 (en) * 1999-07-06 2001-01-11 Surromed, Inc. Bridged fluorescent dyes, their preparation and their use in assays
US6716994B1 (en) * 2000-01-04 2004-04-06 Applera Corporation Mobility-Modifying Cyanine Dyes
US6323337B1 (en) 2000-05-12 2001-11-27 Molecular Probes, Inc. Quenching oligonucleotides
WO2002029003A2 (en) * 2000-10-06 2002-04-11 The Trustees Of Columbia University In The City Of New York Massive parallel method for decoding dna and rna
US6887690B2 (en) * 2001-06-22 2005-05-03 Pe Corporation Dye-labeled ribonucleotide triphosphates
US9261460B2 (en) 2002-03-12 2016-02-16 Enzo Life Sciences, Inc. Real-time nucleic acid detection processes and compositions
US20040161741A1 (en) 2001-06-30 2004-08-19 Elazar Rabani Novel compositions and processes for analyte detection, quantification and amplification
US7166478B2 (en) * 2002-03-12 2007-01-23 Enzo Life Sciences, Inc., C/O Enzo Biochem, Inc. Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses

Also Published As

Publication number Publication date
CA2753910A1 (en) 2003-09-12
US7323571B2 (en) 2008-01-29
IL195306A0 (en) 2009-08-03
US7241897B2 (en) 2007-07-10
IL154700A (en) 2010-11-30
CA2754041A1 (en) 2003-09-12
US20040254355A1 (en) 2004-12-16
IL195307A0 (en) 2009-08-03
JP2009192543A (en) 2009-08-27
CA2754186A1 (en) 2003-09-12
CA2754415C (en) 2016-05-17
JP4551063B2 (en) 2010-09-22
US20040203038A1 (en) 2004-10-14
US20060172308A1 (en) 2006-08-03
US6949659B2 (en) 2005-09-27
US7553959B2 (en) 2009-06-30
CA2754415A1 (en) 2003-09-12
IL195440A (en) 2010-11-30
IL206078A0 (en) 2010-11-30
IL195303A0 (en) 2009-08-03
CA2754185A1 (en) 2003-09-12
IL195303A (en) 2011-07-31
IL195308A0 (en) 2009-08-03
US8357793B2 (en) 2013-01-22
US20040229248A1 (en) 2004-11-18
US20050004350A1 (en) 2005-01-06
IL154700A0 (en) 2003-10-31
EP1348713A2 (en) 2003-10-01
CA2754185C (en) 2017-03-07
EP1348713B1 (en) 2019-01-23
US7256299B2 (en) 2007-08-14
US20040176586A1 (en) 2004-09-09
CA2753922A1 (en) 2003-09-12
IL195436A (en) 2010-11-30
US7863431B2 (en) 2011-01-04
IL195437A (en) 2010-11-30
US20090069500A1 (en) 2009-03-12
US20030225247A1 (en) 2003-12-04
CA2753910C (en) 2016-12-06
IL195305A0 (en) 2009-08-03
EP1348713A3 (en) 2007-07-11
CA2753922C (en) 2016-06-28
IL195435A (en) 2010-11-30
US7163796B2 (en) 2007-01-16
US20110098441A1 (en) 2011-04-28
IL195304A0 (en) 2009-08-03
US7166478B2 (en) 2007-01-23
CA2754186C (en) 2016-04-26
CA2421552C (en) 2011-11-22
US7256291B2 (en) 2007-08-14
JP2010156713A (en) 2010-07-15
CA2754041C (en) 2016-04-26
US20040230036A1 (en) 2004-11-18
JP2004004048A (en) 2004-01-08
IL195308A (en) 2011-04-28
US20040192893A1 (en) 2004-09-30
IL195438A (en) 2010-12-30
US7537751B2 (en) 2009-05-26

Similar Documents

Publication Publication Date Title
CA2421552A1 (en) Labeling reagents and labeled targets, target labeling processes and other processes for using same in nucleic acid determinations and analyses
US6673550B2 (en) Electrophoretic tag reagents comprising fluorescent compounds
US8389701B2 (en) Multisignaling oligomeric or polymeric compositions comprising labeled moieties and binding partners
AU2002367817B2 (en) Bioconjugate-nanoparticle probes
EP0977766A1 (en) Glycoconjugated fluorescent labeling reagents
US20090226940A1 (en) Novel fluorescent dyes and uses thereof
CA2341520C (en) Energy transfer dyes
JP4898119B2 (en) Detectable labeled nucleoside analogues and methods of use thereof
JPH05219998A (en) Method for bonding gene probes by novel anhydrous mixture
US20020142329A1 (en) Compositions and methods employing cleavable electrophoretic tag reagents
US5384241A (en) Specific binding assay compound with inhibitive self-quenching characteristics
WO2000056933A1 (en) Chemically reactive plane-rigidized cyanine dyes and their derivatives
US6967250B1 (en) Energy transfer dyes

Legal Events

Date Code Title Description
EEER Examination request
MKEX Expiry

Effective date: 20230313