Compartmentalised combinatorial chemistry by microfluidic control

• US 9,925,504 B2
• Filed: 12/20/2006
• Issued: 03/27/2018
• Est. Priority Date: 03/31/2004
• Status: Active Grant

• Current Assignee: Medical Research Council (Government of United Kingdom), President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: Medical Research Council (Government of United Kingdom), President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Griffiths, Andrew, Weitz, David, Ahn, Keunho, Link, Darren, Bibette, Jerome
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US11/643,151
• Publication Number:
  US 20070184489A1
• Time in Patent Office: 4,115 Days
• Field of Search: None
• CPC Class Codes: B01F 23/41 Emulsifying
• CPC Class Codes: B01F 25/433 Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
• CPC Class Codes: B01F 25/4331 Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
• CPC Class Codes: B01F 25/4338 Mixers with a succession of converging-diverging cross-sections, i.e. undulating cross-section
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01F 33/3031 using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
• CPC Class Codes: B01J 13/025 Applications of microcapsules not provided for in other subclasses
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B01J 2219/00853 Employing electrode arrangements
• CPC Class Codes: B01J 2219/0086 Dimensions of the flow channels
• CPC Class Codes: B01J 2219/00889 : Mixing micromixers B01F33/30
• CPC Class Codes: B01L 2200/0605 : Metering of fluids
• CPC Class Codes: B01L 2200/0673 : Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0864 : comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 : Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0406 : capillary forces
• CPC Class Codes: B01L 2400/0415 : electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/084 : Passive control of flow resistance
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B82Y 10/00 : Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07K 1/047 : Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C12N 15/1096 : cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
• CPC Class Codes: C12Q 2563/159 : Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components
• CPC Class Codes: C12Q 2600/16 : Primer sets for multiplex assays
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/08 : Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/08 : Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
• CPC Class Codes: G01N 2035/1034 : Transferring microquantities of liquid
• CPC Class Codes: H05K 999/99 : dummy group

Abstract

The invention describes a method for the synthesis of compounds comprising the steps of: (a) compartmentalizing two or more sets of primary compounds into microcapsules; such that a proportion of the microcapsules contains two or more compounds; and (b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets; wherein one or both of steps (a) and (b) is performed under microfluidic control; preferably electronic microfluidic control The invention further allows for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, and which is co-compartmentalized into the microcapsules.

Claims

1. A method comprising:

providing at least a first aqueous fluid comprising at least a first compound,

providing at least a second aqueous fluid comprising at least a second compound,

encapsulating said at least first compound into at least a first aqueous microcapsule within an immiscible oil that comprises a surfactant within a first channel of a microfluidic device,

encapsulating said at least second compound into at least a second aqueous microcapsule within an immiscible oil that comprises a surfactant within a second channel of the microfluidic device,

applying an electric field at a junction of the first channel and the second channel to fuse said at least first and said at least second aqueous microcapsules to form a fused aqueous microcapsule within a third channel of the microfluidic device, thereby causing a chemical reaction between the at least first and at least second compounds to form a fluorogenic reaction product within said fused aqueous microcapsule, and

monitoring for the fluorogenic reaction product by flowing the fused aqueous microcapsule past a detection module that comprises a light source coupled to an optical path used to deliver excitation light to the fused aqueous microcapsule as it passes over the optical path, wherein a plurality of wavelengths of emitted fluorescent light each associated with a different fluorogenic reaction product is collected by the same optical path and the fluorescent light is directed to one or more detectors to measure intensity of each of the wavelengths of the emitted fluorescent light.

2. The method of claim 1, further comprising isolating said fluorogenic reaction product.

3. The method of claim 2, wherein said isolating comprises sorting said fused aqueous microcapsule, comprising said fluorogenic reaction product, within an electric field within a channel of a microfluidic device.

4. The method of claim 1, further comprising identifying said at least first and at least second compounds which reacted to form said fluorogenic reaction product.

5. The method of claim 1, wherein said at least first compound is attached to a microbead.

6. The method of claim 1, wherein said at least second compound is attached to a microbead.

7. The method of claim 1, wherein said at least first compound comprises at least a first label and said at least second compound comprises at least a second label, wherein said at least first and said at least second labels are different.

8. The method of claim 1, wherein said at least first aqueous microcapsule comprises no more than one said at least first compound.

9. The method of claim 1, wherein said at least first aqueous microcapsule comprises a plurality of said at least first compounds.

10. The method of claim 1, wherein said at least second aqueous microcapsule comprises no more than one said at least second compound.

11. The method of claim 1, wherein said at least second aqueous microcapsule comprises a plurality of said at least second compounds.

12. The method according to claim 1, wherein the light source is a laser.

13. The method according to claim 1, wherein the optical path comprises an optical fiber.

14. The method according to claim 1, wherein the fluorescent light is directed to the one or more detectors by dichroic beam splitters.

15. The method according to claim 1, wherein the one or more detectors are one or more photomultiplier tubes.

16. The method according to claim 1, wherein the first compound and the second compound are independently nucleic acids.

17. The method according to claim 1, wherein the first compound and the second compound are independently RNA or DNA.