Compartmentalised screening by microfluidic control

Patent metadata

Field	Value
Publication number	US-10705078-B2
Application number	US-201916453759-A
Country	US
Kind code	B2
Filing date	Jun 26, 2019
Priority date	Oct 12, 2004
Publication date	Jul 7, 2020
Grant date	Jul 7, 2020

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, comprising the steps of: a) compartmentalising the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; and b) identifying the compound which binds to or modulates the activity of the target; wherein at least one step is performed under microfluidic control. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development.

First claim

The invention claimed is:

1. A method for screening a repertoire of compounds for a compound having a desired activity, comprising the steps of:
(a) providing an aqueous fluid comprising the repertoire of compounds;
(b) compartmentalizing the repertoire of compounds into microcapsules by partitioning the aqueous fluid with an immiscible fluid as the aqueous fluid is flowing through a microfluidic channel, such that only one compound from the repertoire of compounds is represented in any one microcapsule, and further wherein each microcapsule comprises a target cell; and
(c) determining an effect of one or more of the repertoire of compounds on the target cell by using a cell-based assay that produces a different reaction product in each microcapsule comprising the target cell and one compound from the repertoire of compounds, wherein a specific reaction product depends on the identity of the compound compartmentalized in each respective microcapsule.

2. The method of claim 1 , wherein the target cells are compartmentalized together with the repertoire of compounds in the microcapsules.

3. The method of claim 1 , wherein the repertoire of compounds are attached to microbeads.

4. The method of claim 3 , wherein the repertoire of compounds are attached to microbeads through one or more cleavable linkers.

5. The method of claim 3 , wherein each microbead comprises a detectable tag.

6. The method of claim 3 , further comprising releasing the compounds from the microbeads.

7. The method of claim 3 , wherein each microbead comprises one of the compounds represented in multiple copies.

8. The method of claim 1 , wherein each of the repertoire of compounds comprises a detectable tag.

9. The method of claim 1 , wherein the immiscible fluid comprises an oil.

10. The method of claim 1 , wherein the cell-based assay involves a gene associated with the target cell.

11. The method of claim 1 , wherein the compartmentalizing step comprises partitioning the aqueous fluid with two counter-propagating streams of the immiscible fluid.

Assignees

• Harvard College
• Res & Innovation Uk

Inventors

• Griffiths Andrew David
• Weitz David A
• Link Darren Roy
• Ahn Keunho
• Bibette Jerome

Classifications

• B01F33/3031

  using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids · CPC title

• B01F33/3021

  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 title

• B01F33/3011

  using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream · CPC title

• B01F23/4145

  Emulsions of oils, e.g. fuel, and water · CPC title

• B01F23/4143

  Microemulsions · CPC title