Selection by compartmentalised screening

Patent metadata

Field	Value
Publication number	US-9857303-B2
Application number	US-201113208614-A
Country	US
Kind code	B2
Filing date	Aug 12, 2011
Priority date	Mar 31, 2003
Publication date	Jan 2, 2018
Grant date	Jan 2, 2018

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.

First claim

The invention claimed is:

1. A water-in-perfluorocarbon oil emulsion library comprising:
(a) a perfluorocarbon oil comprising a fluorinated surfactant; and (b) a plurality of individual aqueous microdroplets encapsulated by the perfluorocarbon oil, each aqueous microdroplet of the plurality of individual aqueous microdroplets comprising;
(i) a subset of non-fluorinated nucleic acids, comprising one or more non-fluorinated nucleic acids from of a repertoire of non-fluorinated nucleic acids, wherein the repertoire of non-fluorinated nucleic acids in a plurality of individual aqueous microdroplets are structurally different;
(ii) a biological target enzyme that is reactive with one or more of the nucleic acids from the repertoire of non-fluorinated nucleic acids; and
(iii) a fluorogenic substrate of the biological target enzyme; wherein each non-fluorinated nucleic acids of the each subset of the repertoire of non-fluorinated nucleic acids is insoluble in the perfluorocarbon oil, and wherein the fluorogenic substrate of the biological target enzyme comprises an optical tag, wherein the optical tag produces a selectable change upon complexation of the fluorogenic substrate with the biological target enzyme.

2. The water-in-perfluorocarbon oil emulsion library according to claim 1 , wherein the aqueous microdroplets further comprise microbeads.

3. The water-in-perfluorocarbon oil emulsion library according to claim 2 , wherein the repertoire of non-fluorinated nucleic acids are releasably attached to the microbeads.

4. The water-in-perfluorocarbon emulsion library according to claim 1 , wherein the biological target enzyme in each of the aqueous microdroplets is structurally different.

5. The water-in-perfluorocarbon oil emulsion library according to claim 1 , wherein each aqueous microdroplet further comprises a member of a binding pair.

6. The water-in-perfluorocarbon oil emulsion library according to claim 5 , wherein a member of a binding pair is an antibody.

7. A water-in-perfluorocarbon oil emulsion library comprising: a perfluorocarbon oil comprising: (a) a fluorosurfactant; and (b) a plurality of individual aqueous microdroplets, wherein each aqueous microdroplet of the plurality of individual aqueous microdroplets is individually encapsulated within the perfluorinated oil and comprises:
(i) multiple copies of a single type of nucleic acid, wherein the single type of nucleic acid is insoluble in the perfluorocarbon oil; and
(ii) a target cell, wherein the target cell comprises a biological target enzyme that is reactive with one or more of the multiple copies of the single type of nucleic acid; and a fluorogenic substrate of the biological target enzyme, wherein the fluorogenic substrate of the biological target enzyme comprises an optical tag, wherein the optical tag produces a selectable change upon complexation of the fluorogenic substrate with the biological target enzyme.

8. The water-in-perfluorocarbon oil emulsion library according to claim 7 , wherein the single type of nucleic acid is attached to a microbead.

9. The water-in-perfluorocarbon oil emulsion library according to claim 8 , wherein the single type of nucleic acid is releasably attached to a microbead.

Assignees

• Griffiths Andrew David
• Abell Chris
• Hollfelder Florian
• Mastrobattista Enrico
• Medical Res Council

Inventors

• Griffiths Andrew David
• Abell Chris
• Hollfelder Florian
• Mastrobattista Enrico

Classifications

• G01N33/5432

  Liposomes or microcapsules · CPC title

• B01J2219/00596

  Solid-phase processes · CPC title

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• B01J2219/00702

  Processes involving means for analysing and characterising the products · CPC title

• C40B50/08

  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 title