Compositions and methods for epitope mapping

• US 20020110843A1
• Filed: 05/10/2001
• Published: 08/15/2002
• Est. Priority Date: 05/12/2000
• Status: Abandoned Application

• Current Assignee: David P. Dumas
• Original Assignee: David P. Dumas
• Inventors: Dumas, David P.
• Application Number:
  US09/853,457
• Publication Number:
  US 20020110843A1
• Time in Patent Office: Days
• US Class Current: 435/7.92
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/0074 Biological products
• 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: G01N 33/6878 in eptitope analysis
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides a composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to the reagent ligands. The ligands can be peptides, oligosaccharides, oligonucleotides, or organic molecules. The invention additionally provides methods of determining an epitope in a sample contacting a composition comprising a diverse population of ligands attached to a solid support and a diverse population of antibodies specifically bound to each of the ligands with a sample; and detecting the antibodies bound to the diverse population of ligands. The invention further provides methods of diagnosing a disease, identifying a potential therapeutic agent, and mapping accessible epitopes of a polypeptide using invention compositions.

Claims

1. A composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to said reagent ligands.

2. The composition of claim 1, wherein each of said reagent ligands is bound to a reagent antibody.

3. The composition of claim 1, wherein said reagent ligands are selected from the group consisting of peptides, oligosaccharides, oligonucleotides, and organic molecules.

4. The composition of claim 1, wherein said reagent ligands are on an array.

5. The composition of claim 1, wherein said reagent antibodies are labeled.

6. The composition of claim 5, wherein said label is a fluorescent label.

7. A composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to a subset of said reagent ligands, wherein an unbound reagent ligand has binding activity for a reagent antibody having specificity for a molecule in a sample.

8. The composition of claim 7, wherein said reagent ligands are selected from the group consisting of peptides, oligosaccharides, oligonucleotides, and organic molecules.

9. The composition of claim 7, wherein said reagent ligands are on an array.

10. The composition of claim 7, wherein said reagent antibodies are labeled.

11. The composition of claim 10, wherein said label is a fluorescent label.

13. The method of claim 12, further comprising the step of identifying which of said reagent ligands is unbound by reagent antibody.

14. The method of claim 12, wherein said reagent ligand unbound by reagent antibody has binding activity for an antibody having specificity for a molecule in said sample.

15. The method of claim 12, wherein said reagent ligands are selected from the group consisting of peptides, oligosaccharides, oligonucleotides, and organic molecules.

16. The method of claim 12, wherein said sample is selected from the group consisting of a cell, a tissue, a body fluid, and an organism.

17. The method of claim 12, wherein said tissue is a biopsy from an individual with a disease.

18. The method of claim 12, wherein said sample is a species of animal or plant.

19. The method of claim 12, wherein said reagent ligands are on an array.

20. The method of claim 12, wherein said reagent antibodies are labeled.

21. The method of claim 20, wherein said label is a fluorescent label.

12. A method of determining an epitope in a sample, comprising:

(a) contacting a composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to said reagent ligands with a sample; and

(b) detecting said reagent antibodies bound to said diverse population of reagent ligands.

22. A method of diagnosing a disease, comprising:

(a) contacting a composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to said reagent ligands with a sample from an individual;

(b) detecting said reagent antibodies bound to said diverse population of reagent ligands; and

(c) identifying which of said reagent ligands is unbound by reagent antibody, wherein a reagent ligand unbound by reagent antibody has binding activity for an antibody having specificity for a molecule associated with said disease.

23. The method of claim 22, wherein said reagent ligands are selected from the group consisting of peptides, oligosaccharides, oligonucleotides, and organic molecules.

24. The method of claim 22, wherein said reagent ligands are on an array.

25. The method of claim 22, wherein said reagent antibodies are labeled.

26. The method of claim 25, wherein said label is a fluorescent label.

28. The method of claim 27, wherein said reagent ligands are selected from the group consisting of peptides, oligosaccharides, oligonucleotides, and organic molecules.

29. The method of claim 27, wherein said reagent ligands are on an array.

30. The method of claim 27, wherein said reagent antibodies are labeled.

31. The method of claim 30, wherein said label is a fluorescent label.

32. The method of claim 27, wherein the reagent antibody displaced from said reagent ligands differing in antibody binding is a potential therapeutic antibody.

27. A method of identifying a potential therapeutic target, comprising:

(a) contacting a composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to said reagent ligands with a sample from an individual having a disease;

(b) detecting reagent antibody binding to said diverse population of reagent ligands;

(c) comparing said reagent antibody binding to said diverse population of reagent ligands to the antibody binding of a normal sample contacted with said composition; and

(d) determining which of said reagent ligands differs in antibody binding between said sample from said individual having a disease and said normal sample, wherein a reagent ligand differing in antibody binding between said samples is a potential therapeutic target.

33. A method of mapping accessible epitopes of a polypeptide, comprising:

(a) contacting a composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent antibodies specifically bound to each of said reagent ligands with a polypeptide;

(b) detecting said reagent antibodies bound to said diverse population of reagent ligands; and

(c) identifying which of said reagent ligands is unbound by reagent antibody, wherein a reagent ligand unbound by reagent antibody has binding activity for an antibody having specificity for a polypeptide epitope accessible to said antibody.

34. The method of claim 33, wherein said reagent ligands are peptides.

35. The method of claim 33, wherein said reagent ligands are on an array.

36. The method of claim 33, wherein said reagent antibodies are labeled.

37. The method of claim 36, wherein said label is a fluorescent label.

39. The method of claim 38, further comprising the step of identifying which of said reagent ligands is unbound by reagent binding molecule.

40. The method of claim 38, wherein said reagent ligand unbound by reagent molecule has binding activity for a binding molecule having specificity for a molecule in said sample.

41. The method of claim 38, wherein said reagent ligands are selected from the group consisting of peptides, oligosaccharides, oligonucleotides, and organic molecules.

42. The method of claim 38, wherein said sample is selected from the group consisting of a cell, a tissue, a body fluid, and an organism.

43. The method of claim 38, wherein said tissue is a biopsy from an individual with a disease.

44. The method of claim 38, wherein said sample is a species of animal or plant.

45. The method of claim 38, wherein said reagent ligands are on an array.

46. The method of claim 38, wherein said reagent binding molecules are labeled.

47. The method of claim 38, wherein said label is a fluorescent label.

38. A method of determining a binding activity in a sample, comprising:

(a) contacting a composition comprising a diverse population of reagent ligands attached to a solid support and a diverse population of reagent binding molecules specifically bound to said reagent ligands with a sample; and

(b) detecting said reagent binding molecules bound to said diverse population of reagent ligands.