Libraries of oligomers labeled with different tags

• US 7,399,846 B2
• Filed: 05/11/2004
• Issued: 07/15/2008
• Est. Priority Date: 05/15/1998
• Status: Expired due to Fees

• Current Assignee: Oxford Gene Technology IP Limited (Sysmex Corporation)
• Original Assignee: Oxford Gene Technology IP Limited (Sysmex Corporation)
• Inventors: Hamilton, Alan Lewis, Southern, Edwin Mellor, Elder, John Kenneth, Shchepinov, Mikhail Sergeevich, Housby, John Nicholas
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US10/843,873
• Publication Number:
  US 20050003408A1
• Time in Patent Office: 1,526 Days
• Field of Search: 536/23.1, 536/25.3, 536/25.32, 536/26.6, 530/391.1, 530/402, 435/6
• US Class Current: 536/23.1
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6872 involving mass spectrometry
• 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 30/10 by measuring physical properties, e.g. mass
• 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 50/10 involving encoding steps
• CPC Class Codes: C40B 50/16 involving encoding steps
• CPC Class Codes: C40B 60/10 for identifying library members
• CPC Class Codes: C40B 70/00 Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes

Abstract

A method of making a set of labelled compounds by the use of a preferably particulate support, comprises dividing the support into lots, performing a different chemical reaction on each lot of the support, e.g. to couple a chemical moiety to that lot of the support, tagging a fraction of each lot of the support with a different label, and combining the said lots of the support. The steps are repeated several times, preferably to build up oligomer molecules carrying labels which identify the nature and position of a monomer unit of the oligomer, and which are releasable from the support. Preferred labels, which are releasable from the compounds by cleavage to provide charged groups for analysis by mass spectrometry, are groups of the trityl (trimethylphenyl) family. Also claimed are libraries of these labels and their use in assays and nucleic acid analysis methods.

Claims

1. A reagent comprising a solid support which carries on its surface oligomer molecules, wherein said oligomer molecules comprise the same sequence wherein the oligomer molecules include some shorter oligomer molecules and at least one shorter oligomer molecule carries a label which identifies the nature and position of a monomer unit of the oligomer molecule and terminates the at least one shorter oligomer molecule so it cannot be extended.

2. The reagent as claimed in claim 1, wherein the solid support is a bead.

3. The reagent as claimed in claim 1, wherein the label is joined by a link that is photocleavable to give a charged species for mass spectrometry.

4. The reagent of claim 1, wherein each label is a group of formula R¹R²R³C—

, where R¹, R² and R³ are the same or different and each is a monocyclic or fused ring aromatic group that is substituted or unsubstituted.

5. The reagent of claim 1, wherein at least one of R¹, R² and R³ carries a substituent selected from C¹-C²⁰ alkoxy or hydrocarbyl either unsubstituted or substituted by carboxylic acid, sulphonic acid, nitro, cyano, hydroxyl, thiol, primary, secondary or tertiary amino, primary or secondary amido, anhydride, carbonyl halide or active ester.

6. The reagent of claim 1, wherein the oligomers are oligonucleotides.

7. A library consisting of a plurality of the reagents of claim 1.

8. The reagent of claim 4, wherein R¹, R² and R³ C—

is a substituted monomethoxytrityl group.

9. The reagent of claim 5, wherein R¹, R² and R³ C—

is a substituted monomethoxytrityl group.

10. A library consisting of a plurality of the reagents of claim 2.

11. A library consisting of a plurality of the reagents of claim 3.

12. A library consisting of a plurality of the reagents of claim 4.

13. A library consisting of a plurality of the reagents of claim 5.

14. A library consisting of a plurality of the reagents of claim 6.

15. A reagent comprising a solid support which carries on its surface oligomer molecules, wherein said oligomer molecules comprise the same sequence wherein the oligomer molecules include different sizes of shorter oligomer molecules and at least one shorter oligomer molecule carries a label which identifies the nature and position of a monomer unit of the oligomer molecule, wherein at least one of each different size of shorter oligomer molecule carries a different label and each label is a group of formula R¹R²R³C—

, where for each different label R¹, R² and R³ are the same or different and each R¹, R² and R³ is monocyclic or a fused ring aromatic group that is substituted or unsubstituted.

16. The reagent as claimed in claim 15, wherein the solid support is a bead.

17. The reagent as claimed in claim 15, wherein the label is joined by a link that is photocleavable to give a charged species for mass spectrometry.

18. The reagent of claim 15, wherein at least one of R¹, R² and R³ carries a substituent selected from C¹-C²⁰ alkoxy or hydrocarbyl either unsubstituted or substituted by carboxylic acid, sulphonic acid, nitro, cyano, hydroxyl, thiol, primary, secondary or tertiary amino, primary or secondary amido, anhydride, carbonyl halide or active ester.

19. The reagent of any claim 15, wherein the oligomers are oligonucleotides.

20. A library consisting of a plurality of the reagents of claim 15.

21. The reagent of claim 18, wherein the R¹, R² and R³ C—

group is a substituted monomethoxytrityl group.

22. The reagent of claim 19, wherein R¹, R² and R³ C—

group is a substituted monomethoxytrityl group.

23. A library consisting of a plurality of the reagents of claim 16.

24. A library consisting of a plurality of the reagents of claim 17.

25. A library consisting of a plurality of the reagents of claim 18.

26. A library consisting of a plurality of the reagents of claim 19.