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Method for producing a biomolecule assay chip

外国特許コード F110003679
整理番号 AF12-01EP
掲載日 2011年7月1日
出願国 欧州特許庁(EPO)
出願番号 07827976
公報番号 2090888
公報番号 2090888
出願日 平成19年11月1日(2007.11.1)
公報発行日 平成21年8月19日(2009.8.19)
公報発行日 平成27年12月23日(2015.12.23)
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
  • 2007JP001197 (2007.11.1) WO
  • 特願2006-297267 (2006.11.1) JP
発明の名称 (英語) Method for producing a biomolecule assay chip
発明の概要(英語) It is an object of the present invention to provide a method for producing a biomolecule assay chip using a microreactor technique, and a chip produced by the method.
The present invention provides: a method for producing a chip on which biomolecules are immobilized in an aligned state, which comprises (a) a step of producing a substrate 1 on which a plurality of biomolecules 1 of a single type are immobilized in an aligned state, (b) a step of 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 the step (a), (c) a step of 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 the step (c) so as to bind the biomolecules 2 onto the substrate 2; and a chip produced by the aforementioned method.
特許請求の範囲(英語) [claim1]
1. A method for producing a chip on which protein-nucleic acid complexes are immobilized in an aligned state, which comprises the following steps (a) to (e): (a) a step of producing a substrate 1 on which a plurality of DNA are immobilized in an aligned state; (b) a step of filling microreactors on a microreactor chip comprising the microreactors at positions overlapping with the sequence positions of DNA immobilized on the substrate 1 produced in the step (a), with reaction reagents for synthesizingmRNA; (c) a step of closely attaching the microreactor chip to the substrate 1 so that the reaction reagents for synthesizing mRNA are allowed to come into contact with DNA, and then carrying out a reaction of synthesizing mRNA 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 mRNA on the substrate 2; and (e)immersing the chip with aligned mRNA on substrate 2 in a solution containing a cell-free translation system for synthesizing the protein-nucleic acid complex.
[claim2]
2. The method according to claim 1, wherein the step of producing "a substrate 1 on which a plurality of DNA are immobilized in an aligned state" described in the step (a) according to claim 1 further comprises the following steps (a) to (c): (a) a step of diluting mixed solutions of DNA and reaction reagents for amplifying DNA, and then filling the microreactors on the microreactor chip with the diluted solutions, so that a single molecule or less of DNA can be present therein in a probability distribution manner; (b) a step of carrying out a reaction of amplifying DNA; and (c) a step of superposing the microreactor chip on the substrate 1 so that the reaction solutions contained in the microreactors on the microreactor chip are allowed to come into contact with the substrate 1, so as to immobilize the amplified DNA on the substrate.
[claim3]
3. The method according to claim 2, wherein the reaction of amplifying DNA is a polymerase chain reaction.
[claim4]
4. The method according to any one of claims 1 to 3, wherein the DNA are those to which linker DNA ligates.
[claim5]
5. The method according to claim 4, wherein puromycin binds to the linker DNA.
[claim6]
6. The method according to any one of claims 1 to 5, wherein it comprises immobilizing avidin on the substrate 1 and thereby biotinylating DNA.
[claim7]
7. The method according to any one of claims 1 to 6, wherein the mRNA is immobilized on the substrate 2 via the puromycin-binding linker DNA.
  • 出願人(英語)
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • NEMOTO NAOTO
  • ICHIKI TAKANORI
  • BIYANI MANISH
国際特許分類(IPC)
欧州特許分類/主・副
  • B01J019/00C
  • L01J219/00C10B2
  • L01J219/00C10B4
  • L01J219/00C4B
  • L01J219/00C4F
  • L01J219/00C4H
  • L01J219/00C4J
  • L01J219/00C4L12
  • L01J219/00C4L2B
  • L01J219/00C4L2F
  • L01J219/00C4L2L4
指定国 Contracting States: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR
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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