IP Library › Granted Patent US 11,371,089
Granted Patent B2
US 11,371,089 · App. 16/387,241 · Granted Jun 28, 2022

Biomolecule analysis method

Inventors: Yoichi Makino (Tokyo, JP); Tomoko Kunitomi (Tokyo, JP)
Assignee: TOPPAN PRINTING CO., LTD.
C12Q1/6858B01L3/502715B01L3/502746C12Q1/686G01N21/6428G01N21/6452B01L2200/0642B01L2200/0673B01L2200/0689B01L2300/0829B01L2300/0851B01L2300/0864B01L2300/0887B01L2300/123B01L2300/161B01L2300/168B01L2400/0487B01L2400/086G01N2021/6439
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Quick Facts
Patent No.
US 11,371,089
App. No.
16/387,241
Granted
Jun 28, 2022
Kind
B2
Abstract

A biomolecule analysis method including: feeding a reagent into a reaction container which includes a plurality of wells and filing the reagent in the plurality of wells, the reagent being for causing an enzymatic reaction with regard to a target substance of analysis; feeding an oil sealing solution over the plurality of wells and sealing the reagent into the plurality of wells with the oil sealing solution so that the plurality of wells become a plurality of independent reaction containers; performing the enzymatic reaction by incubating the reaction container; and detecting a signal amplified by the enzymatic reaction.

Claims (47)

1. A biomolecule analysis method comprising:

feeding a wash buffer into a reaction container including an input portion, an outlet portion, a plurality of wells including a well located between the input portion and the outlet portion, a cover portion over the plurality of wells, and a flow channel located between the plurality of wells and the cover portion, the feeding the wash buffer being from the input portion, to fill, in the plurality of wells, the wash buffer being fed into the reaction container through the flow channel;

feeding a reagent from the input portion of the reaction container into the reaction container to fill the reagent in the plurality of wells through the flow channel, the reagent to cause an enzymatic reaction with regard to a target substance of biomolecule analysis;

feeding an oil sealing solution from the input portion to push a portion of the reagent being outside the plurality of wells out of the outlet portion to cause the oil sealing solution to cover over the plurality of wells through the flow channel and to seal portions of the reagent respectively being inside the plurality of wells in the plurality of wells with the oil sealing solution covering the plurality of wells so that the plurality of wells become a plurality of independent reaction containers sealed from each another;

causing the enzymatic reaction by incubating the reaction container; and

detecting a signal amplified by the enzymatic reaction.

2. The biomolecule analysis method according to claim 1 , wherein

the enzymatic reaction is an isothermal reaction.

3. The biomolecule analysis method according to claim 1 , wherein

a volume of each of the wells is equal to or less than 100 picoliters.

4. The biomolecule analysis method according to claim 1 , wherein

the target substance of the biomolecule analysis is a nucleic acid.

5. The biomolecule analysis method according to claim 4 , wherein

the signal is detected without performing a nucleic acid amplification.

6. The biomolecule analysis method according to claim 1 , wherein

the enzymatic reaction is an invasive cleavage assay.

7. The biomolecule analysis method according to claim 1 , further comprising:

measuring a number of wells giving off the signal.

8. The biomolecule analysis method according to claim 1 , wherein

the enzymatic reaction is performed while an adsorption rate of the target substance of the biomolecule analysis with respect to the wells is reduced.

9. The biomolecule analysis method according to claim 1 , wherein

a number of the target substance of the biomolecule analysis is equal to or less than 1 per well.

10. The biomolecule analysis method according to claim 1 , wherein

the enzymatic reaction is performed while the plurality of wells are covered by the oil sealing solution.

11. The biomolecule analysis method according to claim 1 , wherein

at least one of the reagent or the oil sealing solution includes an adsorption inhibitor.

12. The biomolecule analysis method according to claim 11 , wherein

the adsorption inhibitor is a surfactant.

13. The biomolecule analysis method according to claim 1 , wherein

the wash buffer includes an adsorption inhibitor.

14. The biomolecule analysis method according to claim 13 , wherein

the adsorption inhibitor is a surfactant.

15. A biomolecule analysis method comprising:

feeding a reagent into a reaction container filled with a wash buffer, the reaction container including an input portion, an outlet portion, a plurality of wells including a well located between the input portion and the outlet portion, a cover portion over the plurality of wells, and a flow channel located between the plurality of wells and the cover portion, the feeding the reagent being from the input portion, to fill, in the plurality of wells, the reagent being fed into the reaction container through the flow channel, the reagent to cause an enzymatic reaction with regard to a target substance of analysis; and

feeding an oil sealing solution from the input portion to push a portion of the reagent being outside of the plurality of wells out of the outlet portion to cause the oil sealing solution to cover over the plurality of wells through the flow channel and to seal portions of the reagent respectively being inside the plurality of wells in the plurality of wells with the oil sealing solution covering the plurality of wells so that the plurality of wells become a plurality of independent reaction containers;

causing the enzymatic reaction by incubating the reaction container; and

detecting a signal amplified by the enzymatic reaction.

16. The biomolecule analysis method according to claim 15 , wherein

the wash buffer includes an adsorption inhibitor.

17. The biomolecule analysis method according to claim 16 , wherein

the adsorption inhibitor is a surfactant.

18. The biomolecule analysis method according to claim 15 , wherein

the enzymatic reaction is an isothermal reaction.

19. The biomolecule analysis method according to claim 15 , wherein

a volume of each of the wells is equal to or less than 100 picoliters.

20. The biomolecule analysis method according to claim 15 , wherein

the target substance of analysis is a nucleic acid.

Priority Claims (1)
JP JP2014-017942 · Jan 31, 2014 · national
Continuity (3)
Division 15221143 · Jul 27, 2016
Continuation PCTJP2015052803 · Feb 2, 2015
Related Publication 20190241948A1 · Aug 8, 2019