CA2297589C - Colorants de transfert d'energie a fluorescence accrue - Google Patents

Colorants de transfert d'energie a fluorescence accrue Download PDF

Info

Publication number: CA2297589C
Authority: CA; Canada
Prior art keywords: dye; energy transfer; donor; hydrogen; alkyl
Prior art date: 1996-05-03
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.): Expired - Fee Related

Application number

CA2297589A

Other languages

English (en)

Other versions

CA2297589A1 (fr

Inventor

Linda G. Lee

Sandra L. Spurgeon

Barnett Rosenblum

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.)

Applied Biosystems LLC

Original Assignee

Applied Biosystems LLC

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.)

1996-05-03

Filing date

1997-04-23

Publication date

2010-11-16

1996-05-03 Priority claimed from US08/642,330 external-priority patent/US5863727A/en

1996-10-04 Priority claimed from US08/726,462 external-priority patent/US5800996A/en

1997-04-23 Application filed by Applied Biosystems LLC filed Critical Applied Biosystems LLC

1997-11-03 Publication of CA2297589A1 publication Critical patent/CA2297589A1/fr

2010-11-16 Application granted granted Critical

2010-11-16 Publication of CA2297589C publication Critical patent/CA2297589C/fr

2017-04-23 Anticipated expiration legal-status Critical

Status Expired - Fee Related legal-status Critical Current

Links

239000000975 dye Substances 0.000 title claims abstract description 651
238000012546 transfer Methods 0.000 title claims abstract description 209
239000007850 fluorescent dye Substances 0.000 claims abstract description 83
125000000217 alkyl group Chemical group 0.000 claims abstract description 79
150000001336 alkenes Chemical class 0.000 claims abstract description 64
150000001345 alkine derivatives Chemical class 0.000 claims abstract description 64
125000000524 functional group Chemical group 0.000 claims abstract description 15
150000001993 dienes Chemical class 0.000 claims abstract description 14
125000005647 linker group Chemical group 0.000 claims description 124
108091034117 Oligonucleotide Proteins 0.000 claims description 117
239000013615 primer Substances 0.000 claims description 116
239000002773 nucleotide Substances 0.000 claims description 91
229910052739 hydrogen Inorganic materials 0.000 claims description 90
239000001257 hydrogen Substances 0.000 claims description 89
125000003729 nucleotide group Chemical group 0.000 claims description 82
125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 72
150000002431 hydrogen Chemical class 0.000 claims description 71
238000000034 method Methods 0.000 claims description 69
239000000460 chlorine Substances 0.000 claims description 68
239000001226 triphosphate Substances 0.000 claims description 67
235000011178 triphosphate Nutrition 0.000 claims description 65
-1 3-amino-1-propyn-1-yl Chemical group 0.000 claims description 59
125000003368 amide group Chemical group 0.000 claims description 55
239000002777 nucleoside Substances 0.000 claims description 55
239000003153 chemical reaction reagent Substances 0.000 claims description 53
BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims description 52
ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 51
229910052801 chlorine Inorganic materials 0.000 claims description 51
OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 claims description 50
125000003545 alkoxy group Chemical group 0.000 claims description 50
WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 claims description 48
PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 claims description 48
GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 claims description 48
229910052794 bromium Inorganic materials 0.000 claims description 48
239000011737 fluorine Substances 0.000 claims description 48
229910052731 fluorine Inorganic materials 0.000 claims description 48
150000002825 nitriles Chemical class 0.000 claims description 47
150000003457 sulfones Chemical class 0.000 claims description 46
238000006243 chemical reaction Methods 0.000 claims description 44
150000003833 nucleoside derivatives Chemical class 0.000 claims description 37
230000005284 excitation Effects 0.000 claims description 35
UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 claims description 34
230000004044 response Effects 0.000 claims description 31
150000007523 nucleic acids Chemical class 0.000 claims description 30
239000000203 mixture Substances 0.000 claims description 28
GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims description 22
QDLAGTHXVHQKRE-UHFFFAOYSA-N lichenxanthone Natural products COC1=CC(O)=C2C(=O)C3=C(C)C=C(OC)C=C3OC2=C1 QDLAGTHXVHQKRE-UHFFFAOYSA-N 0.000 claims description 22
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GJCOSYZMQJWQCA-UHFFFAOYSA-N 9H-xanthene Chemical compound C1=CC=C2CC3=CC=CC=C3OC2=C1 GJCOSYZMQJWQCA-UHFFFAOYSA-N 0.000 claims description 19
125000004429 atom Chemical group 0.000 claims description 19
UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 18
125000005605 benzo group Chemical group 0.000 claims description 14
239000007787 solid Substances 0.000 claims description 12
125000001834 xanthenyl group Chemical group C1=CC=CC=2OC3=CC=CC=C3C(C12)* 0.000 claims description 12
125000003835 nucleoside group Chemical group 0.000 claims description 11
239000003155 DNA primer Substances 0.000 claims description 9
230000002255 enzymatic effect Effects 0.000 claims description 8
OAKPWEUQDVLTCN-NKWVEPMBSA-N 2',3'-Dideoxyadenosine-5-triphosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1CC[C@@H](CO[P@@](O)(=O)O[P@](O)(=O)OP(O)(O)=O)O1 OAKPWEUQDVLTCN-NKWVEPMBSA-N 0.000 claims description 6
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SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 claims description 3
HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 claims description 3
NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 claims description 3
125000006239 protecting group Chemical group 0.000 claims description 3
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ARLKCWCREKRROD-POYBYMJQSA-N [[(2s,5r)-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)CC1 ARLKCWCREKRROD-POYBYMJQSA-N 0.000 claims description 2
SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 claims description 2
RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 claims description 2
URGJWIFLBWJRMF-JGVFFNPUSA-N ddTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)CC1 URGJWIFLBWJRMF-JGVFFNPUSA-N 0.000 claims description 2
239000005549 deoxyribonucleoside Substances 0.000 claims description 2
150000004676 glycans Chemical class 0.000 claims description 2
229920000592 inorganic polymer Polymers 0.000 claims description 2
150000002632 lipids Chemical class 0.000 claims description 2
229920000620 organic polymer Polymers 0.000 claims description 2
229920001184 polypeptide Polymers 0.000 claims description 2
239000005017 polysaccharide Substances 0.000 claims description 2
229920001282 polysaccharide Polymers 0.000 claims description 2
108090000765 processed proteins & peptides Proteins 0.000 claims description 2
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239000005547 deoxyribonucleotide Substances 0.000 claims 2
125000001424 substituent group Chemical group 0.000 abstract description 43
VOPWNXZWBYDODV-UHFFFAOYSA-N Chlorodifluoromethane Chemical compound FC(F)Cl VOPWNXZWBYDODV-UHFFFAOYSA-N 0.000 abstract description 23
229910052760 oxygen Inorganic materials 0.000 abstract description 18
QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 17
239000001301 oxygen Substances 0.000 abstract description 17
125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 abstract description 15
229910052717 sulfur Inorganic materials 0.000 abstract description 12
NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 abstract description 11
239000011593 sulfur Substances 0.000 abstract description 11
VILAVOFMIJHSJA-UHFFFAOYSA-N dicarbon monoxide Chemical group [C]=C=O VILAVOFMIJHSJA-UHFFFAOYSA-N 0.000 abstract description 7
JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 45
ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 41
238000003786 synthesis reaction Methods 0.000 description 34
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238000012163 sequencing technique Methods 0.000 description 33
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239000004202 carbamide Substances 0.000 description 6
MGNZXYYWBUKAII-UHFFFAOYSA-N cyclohexa-1,3-diene Chemical compound C1CC=CC=C1 MGNZXYYWBUKAII-UHFFFAOYSA-N 0.000 description 6
HGCIXCUEYOPUTN-UHFFFAOYSA-N cyclohexene Chemical compound C1CCC=CC1 HGCIXCUEYOPUTN-UHFFFAOYSA-N 0.000 description 6
LPIQUOYDBNQMRZ-UHFFFAOYSA-N cyclopentene Chemical compound C1CC=CC1 LPIQUOYDBNQMRZ-UHFFFAOYSA-N 0.000 description 6
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XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 5
NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 5
HDRRAMINWIWTNU-NTSWFWBYSA-N [[(2s,5r)-5-(2-amino-6-oxo-3h-purin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound C1=2NC(N)=NC(=O)C=2N=CN1[C@H]1CC[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HDRRAMINWIWTNU-NTSWFWBYSA-N 0.000 description 5
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229940035893 uracil Drugs 0.000 description 1

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CA2297589A 1996-05-03 1997-04-23 Colorants de transfert d'energie a fluorescence accrue Expired - Fee Related CA2297589C (fr)

Applications Claiming Priority (5)

Application Number	Priority Date	Filing Date	Title
US08/642,330 US5863727A (en)	1996-05-03	1996-05-03	Energy transfer dyes with enhanced fluorescence
US08/642,330		1996-05-03
US08/726,462 US5800996A (en)	1996-05-03	1996-10-04	Energy transfer dyes with enchanced fluorescence
US08/726,462		1996-10-04
CA002203494A CA2203494C (fr)	1996-05-03	1997-04-23	Colorants de transfert d'energie a fluorescence accrue

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US7825237B2 (en)	1996-05-03	2010-11-02	Applied Biosystems, Llc	Oligonucleotides and analogs labeled with energy transfer dyes

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EP0805190B1 (fr)	1999-12-15	Colorants de transfert d'énergie ayant une fluorescence améliorée
US7399854B2 (en)	2008-07-15	Regents labeled with energy transfer dyes
US7888507B2 (en)	2011-02-15	UV excitable fluorescent energy transfer dyes
US20090118485A1 (en)	2009-05-07	Oligonucleotides and analogs labeled with energy transfer dyes
US6218124B1 (en)	2001-04-17	Method for detecting oligonucleotides using UV light source
CA2297589C (fr)	2010-11-16	Colorants de transfert d'energie a fluorescence accrue
US7825237B2 (en)	2010-11-02	Oligonucleotides and analogs labeled with energy transfer dyes