Selection by compartmentalized screening

• US 10,151,698 B2
• Filed: 12/27/2017
• Issued: 12/11/2018
• Est. Priority Date: 03/31/2003
• Status: Active Grant

• Current Assignee: UK Research and Innovation
• Original Assignee: UK Research and Innovation
• Inventors: Griffiths, Andrew David, Abell, Chris, Hollfelder, Florian, Mastrobattista, Enrico
• Primary Examiner(s): Bunker, Amy M
• Application Number:
  US15/855,612
• Publication Number:
  US 20180128745A1
• Time in Patent Office: 349 Days
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• 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 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 33/5432 Liposomes or microcapsules

Abstract

The invention describes a method for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, by compartmentalizing the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsules; and identifying the compound which binds to or modulates the activity of the target. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development.

Claims

1. A method for analyzing compounds, the method comprising:

introducing a first aqueous fluid comprising first compounds, a second aqueous fluid comprising second compounds, and a third aqueous fluid comprising enzymes into a flow of an immiscible oil in a microfluidic channel to form a plurality of aqueous microcapsules surrounded by the immiscible oil, wherein each of the plurality of aqueous microcapsules comprises at least one of the first compound, at least one of the second compound, and at least one of the enzyme;

conducting a first enzyme-catalyzed reaction between the enzyme and the first compound in the plurality of aqueous microcapsules to produce one or more first reaction products as the plurality of aqueous microcapsules are flowing through the microfluidic channel;

conducting a second enzyme catalyzed reaction between the second compound and the one or more first reaction products in the plurality of aqueous microcapsules to produce one or more second reaction products; and

detecting the second reaction product by detecting a change in optical properties in the plurality of aqueous microcapsules.

2. The method according to claim 1, wherein each microcapsule comprises multiple copies of a single type of the first compound.

3. The method according to claim 1, wherein the first compound is a nucleic acid.

4. The method according to claim 3, wherein the nucleic acid is associated with a cell.

5. The method according to claim 1, wherein detecting comprises fluorescently detecting.

6. The method according to claim 5, wherein fluorescently detecting comprises detecting multiple color fluorescence.

7. The method according to claim 1, further comprising attaching the first compound or the second compound to a microbead.

8. The method according to claim 7, wherein attaching occurs prior to the introducing step.

9. The method according to claim 7, wherein attaching occurs after the introducing step and before the first conducting step.

10. The method according to claim 7, wherein attaching comprises covalently attaching.

11. The method according to claim 7, wherein attaching comprises non-covalently attaching.

12. The method according to claim 7, wherein attaching comprises attaching by a cleavable linker.

13. The method according to claim 1, wherein the immiscible oil comprises a surfactant.