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WO2019023551A1 - Biomarqueurs d'immuno-oncologie et leurs procédés d'utilisation - Google Patents

Biomarqueurs d'immuno-oncologie et leurs procédés d'utilisation Download PDF

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Publication number
WO2019023551A1
WO2019023551A1 PCT/US2018/044048 US2018044048W WO2019023551A1 WO 2019023551 A1 WO2019023551 A1 WO 2019023551A1 US 2018044048 W US2018044048 W US 2018044048W WO 2019023551 A1 WO2019023551 A1 WO 2019023551A1
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Prior art keywords
probe
target
region
nanoreporter
molecule
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Inventor
Patrick DANAHER
Sarah WARREN
Erin PIAZZA
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NS Wind Down Co Inc
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Nanostring Technologies Inc
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Publication of WO2019023551A1 publication Critical patent/WO2019023551A1/fr
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B30/00—Methods of screening libraries
    • C40B30/04—Methods of screening libraries by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Definitions

  • This disclosure relates generally to compositions and methods for detection and
  • the disclosure relates to a multiplexable reporter system for labeling a plurality of target molecules with a unique reporter code utilizing compositions comprising a capture probe and a labeled reporter probe, wherein the capture probe and/or reporter probe are associated directly, or indirectly via intermediate oligonucleotide molecules, to a specific target molecule through hybridization.
  • Nucleic acids can be detected and quantified based on their specific polynucleotide sequences.
  • the basic principle underlying existing methods of detection and quantification is the hybridization of a labeled complementary probe sequence to a target sequence of interest in a sample. The formation of a duplex indicates the presence of the target sequence in the sample.
  • the recent development of DNA microarrays has enabled the detection of the presence or absence of thousands of genes in a biological sample in a single experiment.
  • microarray methods still require significant amounts of biological sample, which can be a critical limitation for drug and diagnostic assays that rely upon biological samples with limited supply, such as biopsies of diseased tissues or samples of a discrete cell type.
  • biological samples with limited supply, such as biopsies of diseased tissues or samples of a discrete cell type.
  • the kinetics of hybridization on the surface of a microarray is less efficient than hybridization in small amounts of aqueous solution.
  • methods exist to estimate the amount of nucleic acid present in a sample based on microarray hybridization result microarray technology thus far does not allow for detection of target molecules on an individual level, nor are there microarray-based methods for directly quantifying the amount of target molecule in a given sample.
  • the present disclosure provides a composition comprising a plurality of first probes, wherein a first probe comprises: (i) a first region comprising a first target-specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table 1 A; and (ii) a second region comprising: a first label attachment region hybridized to at least one first RNA molecule, wherein the at least one first RNA molecule is attached to one or more label monomers that emit light constituting a first signal, and an at least second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to at least one second RNA molecule, wherein the at least one second RNA molecule is attached to one or more label monomers that emit light constituting a second signal; and wherein the second region does not overlap with the first region and does not independently base-pair hybridize with the target nucleic acid.
  • composition further comprising a plurality of second probes, wherein each second probe comprises: (i) a first region comprising a second target- specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table 1 A; and (ii) a second region comprising an affinity moiety; and wherein the first target-specific sequence and the second target-specific sequence base-pair hybridize to non-overlapping regions of the same target molecule.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 or a complement thereof.
  • the first region of a second probe can comprise any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the first region of a first probe comprises any one of SEQ ID Nos 1-770 and the first region of a second probe comprises any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the present disclosure provides a method of detecting a target molecule in a biological sample comprising: (i) contacting said sample with a composition of the present disclosure under conditions that allow hybridization of a first target-specific sequence of a first probe to a target molecule, wherein when a first target-specific sequence of a first probe is bound to a target molecule, the identity of the first and second signals emitted from the first probe and the location of the signals relative to each other constitute at least part of a code that identifies the target molecule; and (ii) detecting the code that identifies the target molecule.
  • the present disclosure provides a method of detecting a target molecule in a biological sample comprising: (i) contacting said sample with a composition of the present disclosure under conditions that allow hybridization of the first target-specific sequence of a first probe and the second target-specific sequence of a second probe to a target molecule, wherein when the first target-specific sequence of a first probe and the second target-specific sequence of a second probe are bound to the target molecule, the identity of the first and second signals emitted from the first probe and the location of the signals relative to each other constitute at least part of a code that identifies the target molecule; and (ii) detecting the code that identifies the target molecule.
  • a composition of the present disclosure can comprise at least 50 first probes, or at least 100 first probes, or at least 770 first probes.
  • compositions and methods of the present disclosure when a first probe is hybridized to a target molecule, the identity of the first and second signals emitted from the first probe and the location of the signals relative to each other can constitute at least part of a code that identifies the target molecule.
  • a first probe can further comprise a third, fourth, fifth and at least sixth label attachment region, wherein: (i) the third label attachment region is hybridized to at least one third RNA molecule, wherein the at least one third RNA molecule is attached to one or more label monomers that emit light constituting a third signal; (ii) the fourth label attachment region is hybridized to at least one fourth RNA molecule, wherein the at least one fourth RNA molecule is attached to one or more label monomers that emit light constituting a fourth signal; (iii) the fifth label attachment region is hybridized to at least one fifth RNA molecule, wherein the at least one fifth RNA molecule is attached to one or more label monomers that emit light constituting a fifth signal; (iv) the at least sixth label attachment region is hybridized to at least one sixth RNA molecule, wherein the at least one sixth RNA molecule is attached to one or more label monomers that emit light constituting a sixth signal; and wherein none of the label attachment regions overlap.
  • compositions and methods of the present disclosure when a first probe is bound to a target molecule, the identity of the first, second, third, fourth, fifth and sixth signals emitted from the first probe and the locations of the signals relative to each other can constitute at least part of a code that identifies the target molecule.
  • the present disclosure provides a composition pair comprising a first composition and a second composition, wherein the first composition comprises a first probe and a second probe, wherein a first probe comprises: (i) a first region comprising a first target-specific sequence that independently base-pair hybridizes to a target molecule comprising any of the genes listed in Table 1 A; and (ii) a second region that does not overlap with the first region and does not bind to the target molecule; and wherein a second probe comprises: (i) a first region that hybridizes to the second region of the first probe; and (ii) a second region comprising a first label attachment region hybridized to at least one first RNA molecule, wherein the at least one first RNA molecule is attached to one or more label monomers that emit light constituting a first signal, and an at least second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to at least one second RNA molecule, wherein the at least one second RNA molecule is
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 or a complement thereof.
  • the first region of a third probe can comprise any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 and the first region of a third probe can comprise any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the identity of the first and second signals emitted from the second probe and the location of the signals relative to each other can constitute at least part of a code that identifies the target molecule.
  • the present disclosure provides a kit comprising a composition of the present disclosure and instructions for use.
  • Figure 1 is an illustration of a direct binding (non-tag-based) nanoreporter system utilizing a reporter probe and capture probe.
  • Both reporter and capture probes contain unique target-specific sequences for direct binding to different regions of the target nucleic acid.
  • the reporter probe contains a 6-position fluorescently-labeled reporter code which uniquely "barcodes" the target sequence.
  • the capture probe also contains an affinity moiety, biotin (B).
  • FIG 2 is an illustration of an indirect binding (tag-based) nanoreporter system utilizing a reporter oligo (Oligo A) and a capture oligo (Oligo B) as intermediate probes that bind to the reporter or capture probe.
  • Both Oligo A and Oligo B contain unique target-specific sequences that bind to different regions of the target nucleic acid, and distinct tag sequences that bind to either the capture probe or the reporter probe.
  • the tag sequence in Oligo B can be a universal tag associated with all target-specific capture sequences.
  • the target-specific sequence in Oligo A is associated with a unique tag sequence.
  • Each species of Oligo A carries a unique tag sequence which is associated with a specific reporter code. Under the appropriate hybridization conditions, Oligo A and Oligo B will bind to the target nucleic acid.
  • the universal capture probe will bind to Oligo B, and a specific reporter probe will bind to its associated Oligo A, uniquely barcoding the target sequence.
  • the present disclosure pertains to a multiplexable reporter system and methods for production and use.
  • the present disclosure is based upon the discovery that nanoreporters comprising a reporter and capture probe and probes that bind to the reporter or capture probe and a target molecule can be utilized for the accurate and efficient detection and quantification of a plurality of target molecules in complex mixtures. Moreover, these nanoreporters allow more reliable and reproducible methods for manufacturing nanoreporters.
  • compositions and methods comprise a reporter probe, or a reporter probe and capture probe, for direct binding to the target specific sequence of the same target molecule.
  • These compositions can also be referred to as a standard, or non-tag based nanoreporter system.
  • the reporter and capture reagents e.g., the label attachment regions and attached labels, and affinity moieties
  • Figure 1 the reporter and capture reagents (e.g., the label attachment regions and attached labels, and affinity moieties) are covalently attached to the target-specific regions.
  • Examples of reporter and capture probes related to the non-tag based nanoreporter system, as well as, methods of using these probes is described in U.S. Patent Nos.
  • the present disclosure provides a composition comprising a plurality of first probes, wherein a first probe comprises: (i) a first region comprising a first target-specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table 1 A; and (ii) a second region comprising: a first label attachment region hybridized to at least one first RNA molecule, wherein the at least one first RNA molecule is attached to one or more label monomers that emit light constituting a first signal, and an at least second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to at least one second RNA molecule, wherein the at least one second RNA molecule is attached to one or more label monomers that emit light constituting a second signal; and wherein the second region does not overlap with the first region and does not independently base-pair hybridize with the target nucleic acid.
  • the preceding composition can further comprise a plurality of second probes, wherein each second probe comprises: (i) a first region comprising a second target-specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table 1 A; and (ii) a second region comprising an affinity moiety; and wherein the first target-specific sequence and the second target-specific sequence base-pair hybridize to non-overlapping regions of the same target molecule.
  • the present disclosure provides a composition comprising a plurality of first probes and a plurality of second probes, (a) each first probe comprising: (i) a first region comprising a first target-specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table 1 A; and (ii) a second region comprising: a first label attachment region hybridized to at least one first RNA molecule, wherein the at least one first RNA molecule is attached to one or more label monomers that emit light constituting a first signal, and an at least second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to at least one second RNA molecule, wherein the at least one second RNA molecule is attached to one or more label monomers that emit light constituting a second signal; and wherein the second region does not overlap with the first region and does not independently base-pair hybridize with the target nucleic acid; and (b) each second probe comprising: (i)
  • a first region of a first probe can comprise a first target-specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table IB.
  • a first region of a second probe can comprise a second target-specific sequence that independently base-pair hybridizes to a target molecule comprising at least five of the genes listed in Table IB.
  • the present disclosure also provides a composition comprising a plurality of first probes and a plurality of second probes, (a) each first probe comprising (i) a first region comprising a first target-specific sequence capable of independently base-pair hybridizing to a target molecule comprising at least five of the genes listed in Table 1; and (ii) a second region that does not overlap with the first region and does not bind to the target molecule; (b) each second probe comprising (i) a first region that binds to the second region of said first probe; (ii) an at least first label attachment region which is hybridized to an at least first RNA molecule, wherein the first RNA molecule is attached to one or more label monomers that emit light constituting a first signal; and (iii) an at least second label attachment region, which is non-overlapping to the at least first label attachment region, and which is hybridized to a second RNA molecule, wherein the second RNA molecule is attached to one or more label monomers that emit light constituting a
  • the composition can further comprise a plurality of third probes and a plurality of fourth probes, (a) each said third probes comprising (i) a first region comprising a second target-specific sequence capable of independently base-pair hybridizing to a target molecule comprising at least five of the genes listed in Table 1; (ii) a second region that does not overlap with the first region and does not bind to the target molecule; (b) said fourth probes comprising (i) a first region that binds to the second region of said third probe; and (ii) an affinity moiety; wherein the first target-specific sequence and the second target-specific sequence bind to different regions of the of the same target molecule.
  • the present disclosure also provides a composition pair comprising a first composition and a second composition, wherein the first composition comprises a first probe and a second probe, (a) said first probe comprising (i) a first region comprising a first target-specific sequence capable of independently base-pair hybridizing to any of the genes listed in Table 1; and (ii) a second region that does not overlap with the first region and does not bind to the target molecule; (b) said second probe comprising (i) a first region that binds to the second region of said first probe; and (ii) a second region comprising an at least first label attachment region which is hybridized to a first RNA molecule, wherein the first RNA molecule is attached to one or more label monomers that emit light constituting a first signal; and (iii) an at least second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to a second RNA molecule, wherein the second RNA molecule is attached to one or more label monomers
  • composition comprises a third probe and a fourth probe, (c) said third probe comprising (i) a first region comprising a second target-specific sequence capable of independently base-pair hybridizing to any of the genes listed in Table 1; and (ii) a second region that does not overlap with the first region and does not bind to the target molecule; (d) said fourth probe comprising (i) a first region that binds to the second region of said third probe; and (ii) a second region comprising at least one affinity moiety, wherein the first region does not overlap with the second region; wherein the first target-specific sequence and the second target-specific sequence bind to different regions of the same target molecule, wherein the first and second probes of the first composition cannot bind to the third or fourth probe of the second composition, and wherein when said composition pair is bound to its target molecule, the identity of the first and second signals and their locations relative to each other constitute at least part of a code that identifies the target molecule.
  • the probes of these compositions can comprise two regions, a first region comprising a target-specific sequence and a second region that does not bind to the target. The first region and the second region do not overlap.
  • the second region contains a tag sequence which can bind or hybridize to a region of a reporter probe or a capture probe.
  • a probe that binds or hybridizes to a reporter probe is referred to herein as a reporter oligo (e.g., Oligo A in Figure 2).
  • a probe that binds or hybridizes to a capture probe is referred to herein as a capture oligo (e.g., Oligo B in Figure 2).
  • These compositions can also be referred to as tag-based nanoreporter system.
  • a label attachment region can comprise an artificial nucleic acid sequence.
  • the second region of a first probe, the second region of a second probe, the second region of a third probe, or a second region of a fourth probe can comprise an artificial nucleic acid sequence.
  • An artificial nucleic acid sequence can comprise a non-naturally occurring nucleic acid sequence that has limited to no homology to any sequences found in nature as to avoid off target-binding.
  • compositions of the present invention are capable of detecting, or hybridizing to, any gene within a biomolecular sample
  • the present disclosure is particularly directed to detecting, or hybridizing to, any of the genes listed in Table 1 A.
  • the present disclosure is particularly directed to detecting, or hybridizing to, any of the genes listed in Table IB.
  • ADAM 12 CXCL1 GNLY ILIA PDCD1 TBX21
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 or a complement thereof.
  • the first region of a second probe can comprise any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 and the first region of a second probe can comprise any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1, 2, 5, 6, 13, 14, 15, 16, 24, 28, 30, 31, 34, 36, 38, 41, 46, 54, 70, 78, 81, 83, 95, 96, 108, 1 12, 1 18, 121, 125, 126, 127, 135, 137, 144, 147, 148, 156, 157, 163, 171, 179, 192, 203, 206, 215, 219, 226, 231, 234, 251, 256, 263, 272, 279, 284, 296, 297, 301, 303, 306, 310, 312, 315, 317, 318, 319, 323, 324, 327, 332, 333, 336, 358, 366, 370, 376, 377, 379, 381, 401, 402, 407, 416, 417, 424, 425, 436, 440, 445, 454, 455, 459, 478, 481, 4
  • the first region of a second probe can comprise any one of SEQ ID Nos 771, 772, 775, 776, 783, 784, 785, 786, 794, 798, 800, 801, 804, 806, 808, 81 1, 816, 824, 840, 848, 851, 853, 865, 866, 878, 882, 888, 891, 895, 896, 897, 905, 907, 914, 917, 918, 926, 927, 933, 941, 949, 962, 973, 976, 985, 989, 996, 1001, 1004, 1021, 1026, 1033, 1042, 1049, 1054, 1066, 1067, 1071, 1073, 1076, 1080, 1082, 1085, 1087, 1088, 1089, 1093, 1094, 1097, 1 102, 1 103, 1 106, 1 128, 1 136, 1 140, 1 146, 1 147, 1 149, 1
  • the first region of a first probe can comprise any one of SEQ ID Nos 1, 2, 5, 6, 13, 14, 15, 16, 24, 28, 30, 31, 34, 36, 38, 41, 46, 54, 70, 78, 81, 83, 95, 96, 108, 112, 118, 121, 125, 126, 127, 135, 137, 144, 147, 148, 156, 157, 163, 171, 179, 192, 203, 206, 215, 219, 226, 231, 234, 251, 256, 263, 272, 279, 284, 296, 297, 301, 303, 306, 310, 312, 315, 317, 318, 319, 323, 324, 327, 332, 333, 336, 358, 366, 370, 376, 377, 379, 381, 401, 402, 407, 416, 417, 424, 425, 436, 440, 445, 454, 455, 459, 478, 481, 489, 4
  • the first region of a first probe can comprise any one of SEQ ID Nos 34, 78, 81, 121, 126, 148, 156, 219, 231, 251, 256, 272, 297, 301, 319, 377, 401, 407, 481, 499, 518, 523, 526, 533, 564, 585, 619, 640, 650, 657, 660, 686, 703, 705, or 742 or a complement thereof.
  • the first region of a second probe can comprise any one of SEQ ID Nos 804, 848, 851, 891, 896, 918, 926, 989, 1001, 1021, 1026, 1042, 1067, 1071, 1089, 1147, 1171, 1177, 1251, 1269, 1288, 1293, 1296, 1303, 1334, 1355, 1389, 1410, 1420, 1427, 1430, 1456, 1473, 1475 or 1512 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 34, 78, 81, 121, 126, 148, 156, 219, 231, 251, 256, 272, 297, 301, 319, 377, 401, 407, 481, 499, 518, 523, 526, 533, 564, 585, 619, 640, 650, 657, 660, 686, 703, 705, or 742 or a complement thereof and the first region of a second probe can comprise any one of SEQ ID Nos 804, 848, 851, 891, 896, 918, 926, 989, 1001, 1021, 1026, 1042, 1067, 1071, 1089, 1147, 1171, 1177, 1251, 1269, 1288, 1293, 1296, 1303, 1334, 1355, 1389, 1410, 1420, 1427, 1430, 1456, 1473, 1475 or 1512 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 or a complement thereof.
  • the first region of a third probe can comprise any one of SEQ ID Nos 771- 1540 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1-770 and the first region of a third probe can comprise any one of SEQ ID Nos 771-1540 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 1, 2, 5, 6, 13, 14, 15, 16, 24, 28, 30, 31, 34, 36, 38, 41, 46, 54, 70, 78, 81, 83, 95, 96, 108, 112, 118, 121, 125, 126, 127, 135, 137, 144, 147, 148, 156, 157, 163, 171, 179, 192, 203, 206, 215, 219, 226, 231, 234, 251, 256, 263, 272, 279, 284, 296, 297, 301, 303, 306, 310, 312, 315, 317, 318, 319, 323, 324, 327, 332, 333, 336, 358, 366, 370, 376, 377, 379, 381, 401, 402, 407, 416, 417, 424, 425, 436, 440, 445, 454, 455, 459, 478, 481,
  • the first region of a third probe can comprise any one of SEQ ID Nos , 771, 772, 775, 776, 783, 784, 785, 786, 794, 798, 800, 801, 804, 806, 808, 811, 816, 824, 840, 848, 851, 853, 865, 866, 878, 882, 888, 891, 895, 896, 897, 905, 907, 914, 917, 918, 926, 927, 933, 941, 949, 962, 973, 976, 985, 989, 996, 1001, 1004, 1021, 1026, 1033, 1042, 1049, 1054, 1066, 1067, 1071, 1073, 1076, 1080, 1082, 1085, 1087, 1088, 1089, 1093, 1094, 1097, 1102, 1103, 1106, 1128, 1136, 1140, 1146, 1147, 1149, 1151, 1171, 1172,
  • the first region of a first probe can comprise any one of SEQ ID Nos 1, 2, 5, 6, 13, 14, 15, 16, 24, 28, 30, 31, 34, 36, 38, 41, 46, 54, 70, 78, 81, 83, 95, 96, 108, 112, 118, 121, 125, 126, 127, 135, 137, 144, 147, 148, 156, 157, 163, 171, 179, 192, 203, 206, 215, 219, 226, 231, 234, 251, 256, 263, 272, 279, 284, 296, 297, 301, 303, 306, 310, 312, 315, 317, 318, 319, 323, 324, 327, 332, 333, 336, 358, 366, 370, 376, 377, 379, 381, 401, 402, 407, 416, 417, 424, 425, 436, 440, 445, 454, 455, 459, 478, 481, 489, 4
  • the first region of a first probe can comprise any one of SEQ ID Nos 34, 78, 81, 121, 126, 148, 156, 219, 231, 251, 256, 272, 297, 301, 319, 377, 401, 407, 481, 499, 518, 523, 526, 533, 564, 585, 619, 640, 650, 657, 660, 686, 703, 705, or 742 or a complement thereof.
  • the first region of a third probe can comprise any one of SEQ ID Nos 804, 848, 851, 891, 896, 918, 926, 989, 1001, 1021, 1026, 1042, 1067, 1071, 1089, 1147, 1171, 1177, 1251, 1269, 1288, 1293, 1296, 1303, 1334, 1355, 1389, 1410, 1420, 1427, 1430, 1456, 1473, 1475 or 1512 or a complement thereof.
  • the first region of a first probe can comprise any one of SEQ ID Nos 34, 78, 81, 121, 126, 148, 156, 219, 231, 251, 256, 272, 297, 301, 319, 377, 401, 407, 481, 499, 518, 523, 526, 533, 564, 585, 619, 640, 650, 657, 660, 686, 703, 705, or 742 or a complement thereof and the first region of a third probe can comprise any one of SEQ ID Nos 804, 848, 851, 891, 896, 918, 926, 989, 1001, 1021, 1026, 1042, 1067, 1071, 1089, 1147, 1171, 1177, 1251, 1269, 1288, 1293, 1296, 1303, 1334, 1355, 1389, 1410, 1420, 1427, 1430, 1456, 1473, 1475 or 1512 or a complement thereof.

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  • Physics & Mathematics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne des compositions et des procédés de détection et de quantification de molécules cibles individuelles dans des échantillons biomoléculaires. En particulier, l'invention concerne des compositions marquées et codées comprenant au moins deux sondes hybridées l'une à l'autre qui sont aptes à se lier à des molécules cibles, et à les identifier, sur la base des codes d'étiquette des sondes. La présente invention concerne en outre des procédés de fabrication et d'utilisation de ces compositions. Les compositions peuvent être utilisées dans des applications de diagnostic, de pronostic, de contrôle de qualité et de criblage.
PCT/US2018/044048 2017-07-28 2018-07-27 Biomarqueurs d'immuno-oncologie et leurs procédés d'utilisation Ceased WO2019023551A1 (fr)

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CN114729363A (zh) * 2019-11-15 2022-07-08 诺格拉制药有限公司 Il-34反义剂及其使用方法
WO2022243299A1 (fr) * 2021-05-17 2022-11-24 Nogra Pharma Limited Agents anti-sens il-34 et leurs procédés d'utilisation
US12071623B2 (en) 2021-05-17 2024-08-27 Nogra Pharma Limited IL-34 antisense agents and methods of using same

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Publication number Priority date Publication date Assignee Title
CN114729363A (zh) * 2019-11-15 2022-07-08 诺格拉制药有限公司 Il-34反义剂及其使用方法
EP4058575A1 (fr) * 2019-11-15 2022-09-21 Nogra Pharma Limited Agents anti-sens il-34 et leurs procédés d'utilisation
JP2023503804A (ja) * 2019-11-15 2023-02-01 ノグラ ファーマ リミテッド Ｉｌ－３４アンチセンス薬剤、およびこれを使用する方法
WO2022243299A1 (fr) * 2021-05-17 2022-11-24 Nogra Pharma Limited Agents anti-sens il-34 et leurs procédés d'utilisation
US12071623B2 (en) 2021-05-17 2024-08-27 Nogra Pharma Limited IL-34 antisense agents and methods of using same

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