Compound arrays for sample profiling

• US 8,969,255 B2
• Filed: 11/21/2012
• Issued: 03/03/2015
• Est. Priority Date: 06/19/2009
• Status: Active Grant

• Current Assignee: Arizona Board of Regents (University of Arizona)
• Original Assignee: Arizona Board of Regents (University of Arizona)
• Inventors: Johnston, Stephen Albert, Stafford, Phillip
• Primary Examiner(s): Gross, Christopher M
• Assistant Examiner(s): REYES, CHRISTOPHER R
• Application Number:
  US13/683,778
• Publication Number:
  US 20130143756A1
• Time in Patent Office: 832 Days
• US Class Current: 506/9
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

The invention provides arrays of compound for use in profiling samples. The arrays include compounds bind to components of the samples at relatively low affinities. The avidity of compounds binding to components of the samples can be increased by forming arrays such that multivalent components of the samples (e.g., antibodies or cells) can bind to more than one molecule of a compound at the same time. When a sample is applied to an array under such conditions, the compounds of the array bind to component(s) of the sample with significantly different avidities generating a profile characteristic of the sample.

Claims

1. A method of determining a status of a subject, comprising:

(a) contacting a sample from the subject comprising at least 100 different antibodies, wherein not all of the at least 100 different antibodies are autoantibodies, with an array of immobilized different peptides of between 10-30 amino acids in length, wherein each different peptide has a unique amino acid sequence, each of the immobilized different peptides occupying different features of the array, wherein at least 50 of the immobilized different peptides have dissociation constants of between 1 mM and 1 μ

M to at least one antibody in the sample, and wherein the array comprises at least 100,000 immobilized different peptides;

(b) detecting binding of the immobilized different peptides in the array to antibodies of the sample to obtain a binding profile of the sample, wherein the binding profile is the aggregate effect of the sample binding to the immobilized different peptides occupying different features of the array; and

(c) analyzing the binding profile by comparing the binding profile detected to a reference binding profile to determine the status of the subject.

2. The method of claim 1, wherein different molecules of a same peptide sequence within a feature of the array are spaced less than 6 nm apart.

3. The method of claim 1, further comprising identifying an antibody of the sample that binds to the immobilized different peptides.

4. The method of claim 3, further comprising detecting the identified antibody with a binding partner known to bind the antibody.

5. The method of claim 3, wherein the identified antibody is detected with a plurality of different binding partners known to bind the antibody.

6. The method of claim 4, wherein the binding partner is an antibody to the identified antibody or a peptide known to bind the identified antibody.

7. The method of claim 4, wherein the binding partner is one of the immobilized different peptides bound to the identified antibody in step (b) of claim 1.

8. The method of claim 4, wherein the binding partner is a synbody.

9. The method of claim 1, wherein the sequences of the peptides are randomly selected.

10. The method of claim 1, wherein the immobilized different peptides are selected without regard to the sample.

11. The method of claim 1, wherein the sequences of the immobilized different peptides have less than 90% sequence identity to a protein expressed by a genome.

12. The method of claim 1, wherein the contacting of the sample to the immobilized different peptides on the array is performed in the presence of a potential competitor of binding of the sample to the immobilized different peptides on the array.

13. The method of claim 12, wherein the competitor is a known binding partner of an antibody.

14. The method of claim 12, wherein the sample is a patient sample, and the potential competitor is a protein known to be associated with a disease affecting the patient.

15. The method of claim 1, wherein the sample is a patient sample.

16. The method of claim 15, wherein the patient is known or suspected to be suffering from a disease.

17. The method of claim 15, wherein the patient is known to be at risk of a disease but is not showing symptoms of the disease.

18. The method of claim 17, wherein the disease is an autoimmune disease.

19. The method of claim 17, wherein the disease is an infectious disease.

20. The method of claim 17, wherein the disease is a disease of the central nervous system (CNS).

21. The method of claim 14, wherein the sample is a blood, urine, or CNS sample from the patient.

22. The method of claim 1, wherein antibody(ies) of the sample are labeled prior to contacting with the array.

23. The method of claim 1, wherein binding of the peptides to antibody(ies) of the sample is detected using a secondary antibody.

24. The method of claim 23, wherein the secondary antibody is an isotype-specific antibody.

25. The method of claim 1, wherein binding of the peptides to antibody(ies) of the sample is detected by surface plasmon resonance (SPR) or mass spectrometry.

26. The method of claim 1, further comprising affinity purifying an antibody of the sample using a peptide determined to bind the antibody.

27. The method of claim 1, further comprising washing unbound antibody(ies) of the sample from the array, and dissociating bound antibody(ies) from the array.

28. The method of claim 1, wherein at least 100 of the immobilized different peptides have dissociation constants of between 1 mM and 1 μ

M to at least one antibody in the sample.

29. The method of claim 1, wherein at least 1000 of the immobilized different peptides have dissociation constants of between 1 mM and 1 μ

M to at least one antibody in the sample.

30. The method of claim 1, wherein at least 1% of the immobilized different peptides have dissociation constants of between 1 mM and 1 μ

M to at least one antibody in the sample.

31. The method of claim 1, wherein the subject'"'"'s sample comprises at least one immunoglobulin isotype.

32. The method of claim 31, wherein the immunoglobulin isotype is IgG, IgA, IgM, IgE or combinations thereof.

33. The method of claim 1, wherein the subject'"'"'s sample comprises the antibody repertoire of the subject.