Solid Support Having Ligand Immobilized Thereon By Using Photocleavable Linker

• US 20070212731A1
• Filed: 07/28/2005
• Published: 09/13/2007
• Est. Priority Date: 07/30/2004
• Status: Active Grant

• Current Assignee: Kobe Chemical Genetics, Co., Ltd.
• Original Assignee: Reverse Proteomics Research Institute Company Limited
• Inventors: Tanaka, Akito, Haramura, Masayuki
• Granted Patent: US 7,456,022 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: B01J 20/261 obtained by reactions only involving carbon to carbon unsaturated bonds macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds per se C08F
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/289 bonded via a spacer
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/3251 comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
• CPC Class Codes: B01J 20/3253 comprising a cyclic structure not containing any of the heteroatoms nitrogen, oxygen or sulfur, e.g. aromatic structures
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: Y10T 436/207497 Molecular oxygen

Abstract

The present invention provides a method of analyzing the interaction between a ligand and a target molecule, which uses a solid support wherein the ligand is immobilized via a linker cleavable by photoirradiation, particularly a method of searching and purifying a target molecule. By intervening a linker specifically cleavable by photoirradiation between the ligand molecule and the solid support, release and elution of the target molecule from the solid support without using a ligand or salt at a high concentration can be enabled, and suppression of release and elution of a nonspecific protein can be enabled.

Claims

1. A solid support for the analysis of a specific interaction between a ligand and a target molecule, wherein the ligand is immobilized via a linker cleavable by photoirradiation.

2. The solid support of claim 1, which is used for searching a target molecule.

3. The solid support of claim 1, which is used for purifying a target molecule.

4. The solid support of claim 1, which is represented by the following formula (I) or (II):

5. The solid support of claim 4, wherein R₂ is a hydrogen atom, an optionally substituted alkyl group or a optionally substituted alkoxy group;

R₃ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group;

R₄ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group; and

R₅ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group.

6. The solid support of claim 4, which is represented by the following formula:

7. A compound represented by the following formula (I′

) or (II′

);

8. A linker used for immobilizing a ligand on a solid support, which is represented by the following formula (I′

) or (II′

);

9. The linker of claim 8, which is cleavable by photoirradiation.

10. A solid support for the analysis of a specific interaction between a ligand and a target molecule, wherein the ligand is immobilized via a linker of claim 9.

11. A method of searching a target molecule, which comprises (1) a step of immobilizing a ligand on a solid support via a linker cleavable by photoirradiation, (2) a step of mixing a solid support with the ligand immobilized thereon, which is obtained in the above-mentioned (1), and a sample wherein a target molecule of the ligand is or is not contained, (3) a step of cleaving a ligand from the solid support by irradiation of light, and (4) a step of confirming binding of the target molecule to the ligand.

12. The method of claim 11, wherein the solid support with a ligand immobilized thereon is represented by the following formula (I) or (II):

13. The method of claim 12, wherein R₂ is a hydrogen atom, an optionally substituted alkyl group or a optionally substituted alkoxy group;

R₃ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group;

R₄ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group; and

R₅ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group.

14. The method of claim 12, wherein the solid support is represented by the following formula:

15. A method of purifying a target molecule, which comprises (1) a step of immobilizing a ligand on a solid support via a linker cleavable by photoirradiation, (2) a step of mixing a solid support with the ligand immobilized thereon, which is obtained in the above-mentioned (1), and a sample wherein a target molecule of the ligand is contained, (3) a step of cleaving a ligand from the solid support by irradiation of light, and (4) a step of recove

16. The method of claim 15, wherein the solid support with a ligand immobilized thereon is represented by the following formula (I) or (II):

17. The method of claim 16, wherein R₂ is a hydrogen atom, an optionally substituted alkyl group or a optionally substituted alkoxy group;

R₃ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group;

R₄ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group; and

R₅ is a hydrogen atom, an optionally substituted alkyl group or an optionally substituted alkoxy group.

18. The method of claim 16, wherein the solid support is represented by the following formula: