METHODS FOR DETECTING ANTIBODIES

• US 20190324044A1
• Filed: 06/26/2019
• Published: 10/24/2019
• Est. Priority Date: 03/27/2008
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Kipps, Thomas J., Messmer, Bradley T., Sanchez, Ana B., Kummel, Andrew C., Ruidiaz, Manuel
• Granted Patent: US 10,921,329 B2
• Time in Patent Office: Days
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/577 involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens; G01N33/53 - G01N33/576 take precedence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/6854 Immunoglobulins
• CPC Class Codes: G01N 33/94 involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors

Abstract

Methods for detection of any antibody utilizing a standardized approach applicable to any antibody which provides highly specific assays specific for individual or multiple antibodies. The methods enable improved pharmacokinetic analysis during development and clinical use of antibody-based therapies as well as determination of diagnostic and/or prognostic factors.

Claims

1-20. -20. (Canceled)

21. A complex comprising a therapeutic monoclonal antibody and a peptide, said complex comprising:

a) a therapeutic monoclonal antibody selected from the group consisting of bevacizumab, rituximab, and trastuzumab, wherein the therapeutic monoclonal antibody is not complexed to an epitope of a target protein; and

b) a peptide complexed to said therapeutic monoclonal antibody, said peptide having a length of about 5-40 amino acids and comprising a mimetope recognized by the therapeutic monoclonal antibody, wherein the mimetope comprises a linear sequence of amino acids which is different than a linear sequence of amino acids in the epitope of the target protein.

22. The complex of claim 21, wherein the mimetope binds to the antigen binding site of the therapeutic monoclonal antibody.

23. The complex of claim 21, wherein the therapeutic monoclonal antibody which is not complexed to an epitope of a target protein is present in a biological sample obtained from a subject.

24. The antibody-peptide complex of claim 21, wherein the peptide has a length of about 5-15 amino acids and is attached directly on a solid support.

25. The complex of claim 22, wherein the uncomplexed therapeutic monoclonal antibody is at concentrations from about 50 ng/ml to about 500 ng/ml.

26. The complex of claim 21, wherein the mimetope is identified from a phage-displayed phage library.

27. The complex of claim 21, further comprising a detectable label on the therapeutic monoclonal antibody or the peptide.

28. The complex of claim 27, wherein the detectable label is detectable by a method selected from the group consisting of:

Western blot analysis, flow cytometry, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), competition immunoassay, dual antibody sandwich assay, chemiluminescent assay, bioluminescent assay, fluorescent assay, and agglutination assay.

29. The antibody-peptide complex of claim 21, wherein the antibody is bevacizumab, and the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:

11-14.

30. The antibody-peptide complex of claim 21, wherein the antibody is rituximab, and the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:

4-5.

31. The antibody-peptide complex of claim 21, wherein the antibody is trastuzumab, and the peptide comprises the amino acid sequence of SEQ ID NO:

15.

32. The complex of claim 23, wherein the biological sample is a fluid.

33. The complex of claim 21, wherein the solid support is a bead.

34. The complex of claim 33, wherein the bead is magnetic.

35. The complex of claim 33, wherein the bead comprises magnetic nanoparticles.

36. The complex of claim 35, wherein the magnetic nanoparticles comprise iron oxide (FeO).

37. The complex of claim 33, wherein the bead has a particle size of about 1 μ

m to 50 μ

m.

39. The complex of claim 23, wherein the subject is a human subject.

40. The complex of claim 23, wherein the complex comprises a therapeutic monoclonal antibody which is present at concentrations below 500 ng/ml in the biological sample.