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Biomolecule assay chip

外国特許コード F110003519
整理番号 AF12-01US
掲載日 2011年6月28日
出願国 アメリカ合衆国
出願番号 44780307
公報番号 20100035769
公報番号 8592348
出願日 平成19年11月1日(2007.11.1)
公報発行日 平成22年2月11日(2010.2.11)
公報発行日 平成25年11月26日(2013.11.26)
国際出願番号 JP2007001197
国際公開番号 WO2008053598
国際出願日 平成19年11月1日(2007.11.1)
国際公開日 平成20年5月8日(2008.5.8)
優先権データ
  • 特願2006-297267 (2006.11.1) JP
  • 2007WO-JP01197 (2007.11.1) WO
発明の名称 (英語) Biomolecule assay chip
発明の概要(英語) (US8592348)
A method for producing a chip on which biomolecules are immobilized in an aligned state, comprises (a) producing a substrate 1 on which a plurality of biomolecules 1 of a single type are immobilized in an aligned state, (b) adding reaction reagents for synthesizing biomolecules 2 to microreactors on a microreactor chip comprising the microreactors at positions overlapping with the sequence positions of the biomolecules 1 immobilized on the substrate 1 produced in step (a), (c) closely attaching the microreactor chip to the substrate 1 so that the reaction reagents for synthesizing the biomolecules 2 are allowed to come into contact with the biomolecules 1, so as to synthesize the biomolecules 2 in the microreactors, and (d) superposing the microreactor chip on a substrate 2 after completion of step (c) so as to bind the biomolecules 2 onto the substrate 2; and a chip produced thereby.
特許請求の範囲(英語) [claim1]
1. A method for producing a chip on which biomolecules are immobilized in an aligned state, which comprises the following steps (a) to (e): (a) producing a substrate 1 on which a plurality of biomolecules 1 of a single type are immobilized in an aligned state;
wherein said biomolecules 1 are cDNA;
(b) filling microreactors on a microreactor chip comprising the microreactors at positions overlapping with the sequence positions of the cDNA biomolecules 1 immobilized on the substrate 1 produced in the step (a), with reaction reagents for synthesizing biomolecules 2;
wherein said biomolecules 2 are mRNA;
(c) closely attaching the microreactor chip to the substrate 1 so that the reaction reagents for synthesizing the mRNA biomolecules 2 are allowed to come into contact with the cDNA biomolecules 1, and then carrying out a reaction of synthesizing the mRNA biomolecules 2 in the microreactors so as to synthesize the mRNA biomolecules 2 in reaction solutions contained in the microreactors;
(d) superposing the microreactor chip on a substrate 2 so that the reaction solutions contained in the microreactors on the microreactor chip are allowed to come into contact with the substrate 2 after completion of the step (c), so as to immobilize the mRNA biomolecules 2 on the substrate 2;
wherein linker DNA is bound to said mRNA biomolecules 2 and a compound that specifically binds to a protein is bound to said linker DNA; and
(e) immersing the substrate 2 in reaction reagents for synthesizing biomolecules 3 from the mRNA biomolecules 2 immobilized on the substrate 2;
wherein said biomolecules 3 are protein-nucleic acid complexes in which said protein is bound to said compound.
[claim2]
2. The method of claim 1, wherein step (a) comprises: diluting mixed solutions of cDNA biomolecules 1 and reaction reagents for amplifying the biomolecules, and then filling microreactors on a microreactor chip with the diluted solutions, so that a single molecule or less of the cDNA biomolecules 1 can be present therein in a probability distribution manner; and carrying out a reaction of amplifying the cDNA biomolecules 1.
[claim3]
3. The method according to claim 2, wherein the reaction of amplifying the cDNA biomolecules 1 is a polymerase chain reaction.
[claim4]
4. The method according to claim 2, comprising immobilizing avidin on the substrate 1, biotinylating the cDNA biomolecules 1 and thereby binding said cDNA biomolecules 1 to said substrate 1 via an avidin-biotin bond.
[claim5]
5. The method according to claim 1, the reaction reagents are a solution containing a cell-free translation system and a solution containing a transcriptase.
[claim6]
6. The method according to claim 1 wherein the cDNA biomolecules 1 are immobilized in wells formed on the substrate 1 in an aligned state.
[claim7]
7. The method of claim 1 wherein said cDNA biomolecules 1 are immobilized as an array on said substrate 1 in said aligned state, said cDNA biomolecules 1 having different sequences than each other, each of said protein-nucleic acid complex biomolecules 3 are immobilized on said substrate 2 in a position corresponding to a position of the cDNA biomolecule 1 in said array that encodes said protein-nucleic acid complex biomolecule 3.
[claim8]
8. The method of claim 1 wherein from said substrate 1 on which said cDNA biomolecules 1 are immobilized in an aligned state is prepared a substrate on which said protein-nucleic acid complex biomolecules 3 encoded by the individual cDNAs are immobilized in positions corresponding to the positions of the immobilized cDNAs, without changing the corresponding positions of protein-nucleic acid complex biomolecules 3 on said substrates.
[claim9]
9. The method of claim 1 comprising performing a reverse transcription reaction of the mRNA of said protein-nucleic acid complex biomolecules 3 to DNA by immersing the substrate 2 on which said protein-nucleic acid complex biomolecules 3 are immobilized in reaction solutions that contain reverse transcriptase.
[claim10]
10. The method of claim 1 wherein said compound is puromycin.
  • 発明者/出願人(英語)
  • NEMOTO NAOTO
  • ICHIKI TAKANORI
  • BIYANI MANISH
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
参考情報 (研究プロジェクト等) CREST Establishment of Innovative Manufacturing Technology Based on Nanoscience AREA
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