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다음과 같은 기능을 한번의 로그인으로 사용 할 수 있습니다.
NTIS 바로가기다음과 같은 기능을 한번의 로그인으로 사용 할 수 있습니다.
DataON 바로가기다음과 같은 기능을 한번의 로그인으로 사용 할 수 있습니다.
Edison 바로가기다음과 같은 기능을 한번의 로그인으로 사용 할 수 있습니다.
Kafe 바로가기국가/구분 | United States(US) Patent 등록 |
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국제특허분류(IPC7판) |
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출원번호 | US-0250702 (2011-09-30) |
등록번호 | US-9562897 (2017-02-07) |
발명자 / 주소 |
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출원인 / 주소 |
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대리인 / 주소 |
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인용정보 | 피인용 횟수 : 0 인용 특허 : 422 |
The invention generally relates to performing sandwich assays in droplets. In certain embodiments, the invention provides methods for detecting a target analyte that involve forming a compartmentalized portion of fluid including a portion of a sample suspected of containing a target analyte and a sa
The invention generally relates to performing sandwich assays in droplets. In certain embodiments, the invention provides methods for detecting a target analyte that involve forming a compartmentalized portion of fluid including a portion of a sample suspected of containing a target analyte and a sample identifier, a first binding agent having a target identifier, and a second binding agent specific to the target analyte under conditions that produce a complex of the first and second binding agents with the target analyte, separating the complexes, and detecting the complexes, thereby detecting the target analyte.
1. A method for measuring the quantity of target analytes, the method comprising: forming a plurality of fluidic compartmentalized portions, each comprising: a target analyte from a sample, a first binding agent specific for a first binding region on the target analyte and comprising a target identi
1. A method for measuring the quantity of target analytes, the method comprising: forming a plurality of fluidic compartmentalized portions, each comprising: a target analyte from a sample, a first binding agent specific for a first binding region on the target analyte and comprising a target identifier, and a second binding agent specific for a second binding region on the target analyte, wherein the second binding agent comprises a functionalized portion for immobilization to a solid support;producing in the one or more compartmentalized portions, one or more complexes each comprising the first and second binding agents bound to the target analyte;disrupting and releasing contents of the plurality of compartmentalized portions;immobilizing the one or more complexes on a solid support through the functionalized portion of the second binding agent to form one or more immobilized complexes;separating the immobilized complexes from the other released contents; anddetecting the target identifiers of the separated immobilized complexes, thereby identifying the quantity of the target analyte. 2. The method according to claim 1, wherein one or more of the plurality of the compartmentalized portions comprise a third binding agent, wherein the third binding agent comprises a second target identifier and is specific to a different target analyte. 3. The method according to claim 1, wherein each of the plurality of compartmentalized portions is a droplet. 4. The method according to claim 3, wherein the droplet is in a microchannel. 5. The method according to claim 3, wherein the droplet is surrounded by an immiscible carrier fluid. 6. The method according to claim 5, wherein the carrier fluid is an oil. 7. The method according to claim 1, wherein the forming step comprises merging a droplet comprising the first and/or the second binding agents with a droplet comprising the target analyte. 8. The method according to claim 1, wherein the forming step comprises flowing a droplet comprising the first and/or the second binding agents such that it interacts with a fluid stream comprising the target analyte, thereby a portion of the fluid stream is integrated with the droplet. 9. The method according to claim 1, wherein each of the compartmentalized portions further comprises a competitive binding inhibitor. 10. The method according to claim 1, wherein the target identifier is releasably attached to the first binding agent. 11. The method of claim 1, wherein the functionalized portion of the second binding agent is a terminal portion of the second binding agent. 12. The method according to claim 1, wherein the separating step comprises: washing the released contents. 13. The method according to claim 1, wherein each of the plurality of compartmentalized portions further comprises an identifier of the sample. 14. The method of claim 13, wherein the identifier of the sample is capable of complexing with the target identifier. 15. The method according to claim 1, wherein the target identifier of each of the plurality of compartmentalized portions is a barcode oligonucleotide. 16. The method according to claim 15, wherein the barcode oligonucleotides are released from the first binding agents after the complexes have been separated. 17. The method of claim 16, wherein the barcode oligonucleotides are amplified. 18. The method according to claim 16, wherein a plurality of the released barcode oligonucleotides are attached to each other to form a nucleic acid strand. 19. The method according to claim 18, wherein the detecting step comprises detecting the nucleic acid strand. 20. The method of claim 19, wherein the detecting step comprises sequencing the nucleic acid strand. 21. The method according to claim 20, wherein sequencing is sequencing-by-synthesis. 22. The method according to claim 21, wherein sequencing-by-synthesis is single molecule sequencing-by-synthesis. 23. A method for a determining a number of a target nucleic acid, the method comprising: forming a plurality of fluidic compartmentalized portions, each comprising: a sample containing a target nucleic acid, wherein the target nucleic acid comprises a binding region and a second binding region, anda first binding agent specific for the first binding region on the target nucleic acid and comprising a target identifier, and a second binding agent specific for the second binding region on the target nucleic acid, wherein the second binding agent comprises a functionalized portion for immobilization to a solid support;producing in the one or more compartmentalizing portions, one or more complexes each comprising the first and second binding agents bound to the target nucleic acid;disrupting and releasing contents of the plurality of compartmentalized portions;immobilizing the one or more complexes on a solid support through the functionalized portion of the second binding agent to form one or more immobilized complexes;separating each immobilized complex fromt the other released contents; anddetecting the target identifiers of the separated immobilized complexes, thereby identifying the target nucleic acid. 24. The method of claim 23, wherein the first binding region of the target nucleic acid is the transcriptional start region and the second binding region of the target nucleic acid is a splice variant region, and the first binding agent and second binding agent are selective for detection of the splice variant. 25. The method of claim 23, wherein the first binding region of the target nucleic acid is a first gene and the second binding region of the target nucleic acid is a second gene, the first and second genes are joined at a fusion junction. 26. The method of claim 23, wherein the first binding region of the target nucleic acid comprises a single nucleotide polymorphism and the second binding region of the target nucleic acid comprises a portion of wild-type sequences in proximity to the single nucleotide polymorphism. 27. The method according to claim 23, wherein the target identifier of each of the one or more compartmentalized portions is a barcode oligonucleotide. 28. The method according to claim 27, wherein the barcode oligonucleotides are released from the first binding agents after the separating step. 29. The method according to claim 28, wherein a plurality of the released barcode oligonucleotides are attached to each other to form a template nucleic acid strand. 30. The method according to claim 29, wherein the nucleic acid strand is amplified. 31. The method according to claim 29, wherein the detecting step comprises sequencing the nucleic acid strand. 32. The method according to claim 31, wherein sequencing is sequencing-by-synthesis. 33. The method according to claim 32, wherein sequencing-by-synthesis is single molecule sequencing-by-synthesis. 34. A method for identifying a number of target biological molecules, the method comprising: forming a plurality of fluidic compartmentalized portions, each comprising: a portion of a sample suspected of containing a target biological molecule and unspecific analytes, wherein the target biological molecule comprises a first binding region and a second binding region,a first binding agent specific for the first binding region on the target biological molecule and comprising a target identifier, and a second binding agent specific for the second binding region on the target biological molecule, and a competitive inhibitor specific to the unspecific analytes, wherein the second binding agent comprises a functionalized portion for immobilization to a solid supportproducing in the one or more compartmentalizing portions, one or more complexes each comprising the first and second binding agents bound to the target molecule;disrupting and releasing contents of the plurality of compartmentalized portions;immobilizing the one or more complexes on a solid support through the functionalized portion of the second binding agent to form one or more immobilized complexes;separating the immobilized complexes from the other released contents; anddetecting the target identifiers of the separated immobilized complexes, thereby identifying the target biological molecules. 35. The method of claim 34, wherein each of the plurality of compartmentalized portions is a droplet. 36. The method according to claim 35, wherein the droplet is present in a microchannel. 37. The method according to claim 35, wherein the droplet is surrounded by an immiscible carrier fluid. 38. The method according to claim 37, wherein the carrier fluid is oil. 39. The method according to claim 26, wherein the compartmentalized portion further comprises a sample identifier that binds the target identifier. 40. The method according to claim 39, wherein the target identifier of each of the plurality of compartmentalized portions is a barcode oligonucleotide. 41. The method according to claim 40, wherein the barcode oligonucleotides are released from the first binding agents after the separating step. 42. The method according to claim 41, wherein a plurality of the released barcode oligonucleotides are attached to each other to form a template nucleic acid strand. 43. The method according to claim 42, wherein the nucleic acid strand is amplified. 44. The method according to claim 43, wherein the detecting step comprises sequencing the nucleic acid strand. 45. The method according to claim 1, wherein the target analyte comprises a plurality of members and at least two members are coupled together prior to the producing the complex. 46. The method according to claim 1, wherein the target analyte comprises a nucleic acid member complexed to a protein member. 47. The method according to claim 1, wherein the target analyte comprises a first protein member complexed to a second protein member. 48. The method according to claim 34, wherein the target biological molecules comprise at least two members coupled together prior to the producing the complex. 49. The method according to claim 34, wherein the target biological molecules comprise a nucleic acid member complexed to a protein member. 50. The method according to claim 34, wherein the target biological molecules comprise a first protein member complexed to a second protein member. 51. The method of claim 1 wherein the target identifiers are released from the first binding agents after the separating step. 52. The method of claim 51 wherein the target identifiers are released from the complexes using a process selected from the group consisting of light induced release, enzymatic release, chemical induced release, and temperature induced release. 53. The method of claim 52 wherein the process of light induced release comprises UV photocleavage. 54. The method of claim 1, wherein each of the plurality of compartmentalized portions further comprises a third binding agent specific to a third binding region of the target analyte and comprising a second target identifier. 55. The method of claim 19, wherein the detecting step comprises digital PCR. 56. The method of claim 29, wherein the detecting step comprises digital PCR. 57. The method of claim 43, wherein the detecting step comprises digital PCR. 58. The method according to claim 34, further comprising the step of counting the target identifiers of the separated immobilized complexes. 59. The method according to claim 34, wherein two or more of the compartmentalized portions comprise a third binding agent having affinity to a different target biological molecule comprising one or more members and comprising a second target identifier and a fourth binding agent having affinity to the different target. 60. The method according to claim 34, wherein the forming step comprises merging a droplet comprising the first and/or the second binding agents with a droplet comprising a target biological molecule. 61. The method according to claim 34, wherein the forming step comprises flowing a droplet comprising the first and/or the second binding agents such that it interacts with a fluid stream comprising one or more target biological molecules, thereby a portion of the fluid stream is integrated with the droplet. 62. The method according to claim 34, wherein the target identifier is releasably attached to the first binding agent. 63. The method of claim 34, wherein the functionalized portion of the second binding agent is a terminal portion of the second binding agent. 64. The method according to claim 34, wherein the separating step comprises: washing the released contents. 65. The method according to claim 34, wherein the compartmentalized portion further comprises an identifier of the sample. 66. The method of claim 65, wherein the identifier of the sample is capable of complexing with the target identifier. 67. The method according to claim 41, wherein the barcode oligonucleotides are amplified. 68. The method according to claim 42, wherein the detecting step comprises detecting the nucleic acid strand. 69. The method according to claim 44, wherein sequencing is sequencing-by-synthesis. 70. The method according to claim 69, wherein sequencing-by-synthesis is single molecule sequencing-by-synthesis. 71. The method of claim 23, wherein the first and second binding regions of the target nucleic acid are two splice variant regions. 72. The method according to claim 1, wherein the target analyte comprises a modified analyte. 73. The method according to claim 1, wherein the target analyte comprises a post-translational protein. 74. The method according to claim 1, wherein the forming step comprises merging a plurality of compartmentalized portions each comprising the first and/or the second binding agents with a compartmentalized portion comprising the target analyte. 75. The method according to claim 74, further comprising forming a plurality of compartmentalized portions each comprising the first and/or the second binding agents. 76. The method according to claim 34, wherein the forming step comprises merging a plurality of compartmentalized portions each comprising the first and/or the second binding agents with a compartmentalized portion comprising one or more target biological molecules. 77. The method according to claim 76, further comprising forming a plurality of compartmentalized portions each comprising the first and/or the second binding agents.
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