Method of synthesis and testing of combinatorial libraries using microcapsules

• US 20060154298A1
• Filed: 09/29/2005
• Published: 07/13/2006
• Est. Priority Date: 03/31/2003
• Status: Active Grant

• Current Assignee: UK Research and Innovation
• Original Assignee: Medical Research Council (Government of United Kingdom)
• Inventors: Griffiths, Andrew David, Abell, Chris, Mastrobattista, Enrico, Hollfelder, Florian
• Granted Patent: US 7,718,578 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• 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 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• 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 2219/00835 Comprising catalytically active material
• CPC Class Codes: B01J 2219/0086 Dimensions of the flow channels
• CPC Class Codes: B01J 2219/00894 More than two inlets
• CPC Class Codes: B01J 2219/00903 Segmented flow
• 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: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 1/047 Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C12N 15/1075 : by coupling phenotype to genotype, not provided for in other groups of this subclass
• CPC Class Codes: C40B 20/04 : Identifying library members by means of a tag, label, or other readable or detectable entity associated with the library members, e.g. decoding processes
• CPC Class Codes: C40B 30/04 : by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• 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 33/5432 : Liposomes or microcapsules

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. 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 for preparing a repertoire of compounds comprising the steps of:

(a) compartmentalising two or more sets of primary compounds into microcapsules;

such that a proportion of the microcapsules contains multiple copies of one or more compounds representative of each of said sets, and wherein said one or more compounds form a subset of the set of primary compounds;

(b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets.

2. A method according to claim 1, wherein the subset of primary compounds constitutes 10% or less of the set of primary compounds.

3. A method according to claim 2, wherein the subset of primary compounds constitutes 1% or less of the set of primary compounds.

4. A method according to claim 3, wherein the subset of primary compounds is a single compound.

5. A method according to any preceding claim, wherein, in step (a), the modal number of different primary compounds per compartment will be equivalent to the number of primary compounds forming the secondary compound in step (b).

6. A method according to any preceding claim, wherein step (a) comprises forming separate emulsion compartments comprising primary compounds and mixing the emulsion compartments to form an emulsified set of primary compounds wherein a subset of the set of primary compounds is represented in multiple copies in any one microcapsule.

7. A method according to any preceding claim, wherein step (a) comprises

a) attaching the set of primary compounds onto microbeads, such that only a subset of the set of primary compounds is represented on any one microbead;

and

b) compartmentalising the microbeads into microcapsules;

such that a subset of the set of primary compounds is represented in multiple copies in any one microcapsule.

65. Method according to claim 1, wherein the compound or compounds are low molecular weight compounds.

67. A method according to claim 1, wherein microencapsulation is achieved by forming a water-in-oil emulsion.

94. A method according to claim 7, wherein the primary compounds are coupled to beads with different optical properties.

101. A product when isolated according to the method of claim 1.

102. A method according to claim 1, further comprising the step of isolating the secondary compound produced by reaction of the primary compounds.

8-13. -13. (canceled)

14. A method for identifying primary compounds which react together to form secondary compounds capable of binding to or modulating the activity of a target, comprising the steps of:

(a) compartmentalising two or more sets of primary compounds into microcapsules;

such that a proportion of the microcapsules contains two or more compounds;

(b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets; and

(c) identifying subsets of primary compounds which react to form secondary compounds which bind to or modulate the activity of the target.

18. A method according to claim 14, wherein the activity of the target is a binding activity.

19. A method according to claim 14, wherein the activity of the target is a catalytic activity.

20. A method according to claim 14, wherein the secondary compound reacts with the target to generate a reaction product.

21. A method according to claim 14, wherein the target is compartmentalised into microcapsules together with the compounds.

22. A method according to claim 14 wherein a scaffold molecule is compartmentalised into microcapsules together with the compounds.

24. A method according to claim 14, wherein the compound having a desired activity causes a change in the microcapsule or microbead which allows the microcapsule or microbead to be identified, sorted or selected.

36. A method according to claim 14, wherein the compound having a desired activity causes a change in the microcapsule which allows the microcapsule to be identified, sorted or selected.

37. A method according to claim 14, wherein the modification of the target within the microcapsule induces a change in the optical properties of the microcapsule.

48. A method according to claim 24, wherein the compound in a microcapsule is identified by a difference in the optical properties of the microcapsule.

83. A method according to claim 24, wherein the microcapsules are identified and, optionally, sorted by detection of a change in their fluorescence.

15. A method for synthesising compounds with enhanced ability to bind to or modulate the activity of the target, comprising the steps of:

(a) compartmentalising into microcapsules subsets of primary compounds identified in step (c) of the second aspect of the invention and, optionally, compartmentalising additional sets of primary compounds;

(b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets; and

(c) identifying subsets of primary compounds which react to form secondary compounds which bind to or modulate the activity of the target.

16. (canceled)

17. A method for identifying individual compounds which bind to or modulate the activity of the target, comprising the steps of:

(a) compartmentalising into microcapsules a primary compound identified in step (c) of the second or third aspect of the invention and additional sets of primary compounds;

(b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets; and

(c) identifying subsets of primary compounds which react to form secondary compounds which bind to or modulate the activity of the target.

23. (canceled)

25-35. -35. (canceled)

38-47. -47. (canceled)

49-64. -64. (canceled)

66. (canceled)

68-82. -82. (canceled)

84-93. -93. (canceled)

95-100. -100. (canceled)

103-110. -110. (canceled)