Method for detecting target molecule

Patent metadata

Field	Value
Publication number	US-2016223531-A1
Application number	US-201615082195-A
Country	US
Kind code	A1
Filing date	Mar 28, 2016
Priority date	Mar 8, 2011
Publication date	Aug 4, 2016
Grant date	—

Abstract

This invention provides a technique enabling to detect target molecules of low concentration with high sensitivity. This invention includes (i) a step of introducing a hydrophilic solvent ( 42 ) containing beads ( 40 ),( 41 ′) into a space ( 30 ) between (a) a lower layer section ( 10 ) including a plurality of receptacles ( 13 ) each of which is capable of storing only one of the beads ( 41 ),( 41 ′) and which are separated from each other by a side wall ( 12 ) having a hydrophobic upper surface and (b) an upper layer section ( 20 ) facing a surface of the lower layer section ( 10 ) on which surface the plurality of receptacles ( 13 ) are provided; and (ii) a step of introducing a hydrophobic solvent ( 43 ) into the space ( 30 ), the step (ii) being carried out after the step (i).

First claim

1 - 12 . (canceled)

13 . A method for detecting a target molecule, comprising:
(i) a step of reacting beads having an average particle diameter of 1 μm to 4 μm and specifically capturing target molecules with the target molecules; (ii) a step of introducing a hydrophilic solvent containing the beads having the average particle diameter of 1 μm to 4 μm into a space within a flow cell structure, the space being between
(a) a lower layer section including a plurality of receptacles each of which has a size that can store only one of the beads and which are separated from each other by a side wall having a hydrophobic upper surface and
(b) an upper layer section facing a surface of the lower layer section on which surface the plurality of receptacles are provided,
the step (ii) being carried out after the step (i); (iii) a step of introducing a hydrophobic solvent into the space to displace the hydrophilic solvent, the step (iii) being carried out after the step (ii) to form, in the plurality of receptacles, droplets of the hydrophilic solvent within the plurality of receptacles covered with the hydrophobic solvent, wherein
a single bead is trapped within a droplet of the hydrophilic solvent within each of the plurality of receptacles,
the plurality of receptacles each have a width that is 1.5 to 2 times larger than the average particle diameter of the beads,
the plurality of receptacles each have a depth that is equal to or smaller than 1.5 times the average particle diameter of the beads, and
a trapping efficiency of the beads is twenty-five times or more higher relative to the absence of the flow cell structure, wherein the structure is lacking an upper layer section; and
(iv) a step of determining whether or not any one of the beads having captured the target molecules is stored in each of the plurality of receptacles, the step (iv) being carried out after the step (iii).

14 . The method as set forth in claim 13 , wherein:
the beads are such beads to which molecules specifically bindable to the target molecules are bound.

15 . The method as set forth in claim 13 , further comprising:
(v) a step of deaerating the space,
the step (v) being carried out after the step (ii) and before the step (iii).

16 . The method as set forth in claim 14 , further comprising:
(v) a step of deaerating the space,
the step (v) being carried out after the step (ii) and before the step (iii).

17 . The method as set forth in claim 13 , wherein:
the hydrophilic solvent is at least one selected from the group consisting of water, hydrophilic alcohol, hydrophilic ether, ketone, nitrile solvents, dimethyl sulfoxide, and N,N-dimethylformamide, or is a mixture including the at least one.

18 . The method as set forth in claim 14 , wherein:
the hydrophilic solvent is at least one selected from the group consisting of water, hydrophilic alcohol, hydrophilic ether, ketone, nitrile solvents, dimethyl sulfoxide, and N,N-dimethylformamide, or is a mixture including the at least one.

19 . The method as set forth in claim 13 , wherein:
the hydrophobic solvent is at least one selected from the group consisting of saturated hydrocarbon, unsaturated hydrocarbon, aromatic hydrocarbon, silicone oil, perfluorocarbon, halogen solvents, and hydrophobic ionic liquid, or is a mixture including the at least one.

20 . The method as set forth in claim 14 , wherein:
the hydrophobic solvent is at least one selected from the group consisting of saturated hydrocarbon, unsaturated hydrocarbon, aromatic hydrocarbon, silicone oil, perfluorocarbon, halogen solvents, and hydrophobic ionic liquid, or is a mixture including the at least one.

Assignees

• Japan Science & Tech Agency

Inventors

• Noji Hiroyuki
• Iino Ryota
• Araki Suguru

Classifications

• G01N33/54313

  the carrier being characterised by its particulate form · CPC title

• B01L2300/161

  Control and use of surface tension forces, e.g. hydrophobic, hydrophilic · CPC title

• G01N33/54306 Primary

  Solid-phase reaction mechanisms · CPC title

• B01L2300/0858

  Side walls · CPC title

• B01L2200/0668

  Trapping microscopic beads · CPC title