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Biomolecule assay chip

Foreign code F110003519
File No. AF12-01US
Posted date Jun 28, 2011
Country United States of America
Application number 44780307
Gazette No. 20100035769
Gazette No. 8592348
Date of filing Nov 1, 2007
Gazette Date Feb 11, 2010
Gazette Date Nov 26, 2013
International application number JP2007001197
International publication number WO2008053598
Date of international filing Nov 1, 2007
Date of international publication May 8, 2008
Priority data
  • P2006-297267 (Nov 1, 2006) JP
  • 2007WO-JP01197 (Nov 1, 2007) WO
Title Biomolecule assay chip
Abstract (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.
Scope of claims [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.
  • Inventor, and Inventor/Applicant
  • NEMOTO NAOTO
  • ICHIKI TAKANORI
  • BIYANI MANISH
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) CREST Establishment of Innovative Manufacturing Technology Based on Nanoscience AREA
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