Compartmentalised screening by microfluidic control
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) 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 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.
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) compartmentalising 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 a subset of the repertoire is represented in multiple copies in any one microcapsule and wherein each microcapsule comprises a target cell;
(c) conducting a reaction involving a molecule associated with the target cell, wherein the compound has an effect on the reaction; and
(d) detecting a product of the reaction to identify a microcapsule containing the compound having the desired activity.
2. The method of claim 1 , wherein the target cell is compartmentalized together with the repertoire of compounds in the microcapsules.
3. The method of claim 1 , further comprising fusing the microcapsules with second microcapsules comprising the target cells.
4. The method of claim 1 , wherein the repertoire of compounds are attached to microbeads.
5. The method of claim 4 , wherein each microbead comprises a detectable tag.
6. The method of claim 4 , wherein the repertoire of compounds are attached to microbeads through one or more cleavable linkers.
7. The method of claim 4 , further comprising releasing the compounds from the microbeads.
8. The method of claim 4 , wherein each microbead comprises one of the subsets of the repertoire of compounds represented in multiple copies.
9. The method of claim 1 , wherein the molecule associated with the target cell is an enzyme or a receptor.
10. The method of claim 9 , further comprising incubating the microcapsules.
11. The method of claim 1 , wherein the target cell is from a single cell organism.
12. The method of claim 1 , wherein the target cell is from a multicellular organism.
13. The method of claim 1 , wherein each of the repertoire of compounds comprises a detectable tag.
14. The method of claim 1 , wherein the immiscible fluid comprises a fluorocarbon oil.
15. The method of claim 1 , wherein the molecule associated with the target cell is a gene, and wherein at least one compound from the repertoire of compounds expresses or inhibits expression of a gene.