Anti-SOX10 Antibody Systems and Methods

• US 20160216269A1
• Filed: 10/03/2014
• Published: 07/28/2016
• Est. Priority Date: 10/03/2013
• Status: Active Grant

• Current Assignee: Biocare Medical LLC
• Original Assignee: Biocare Medical LLC
• Inventors: Tacha, David, Qi, Weimin
• Granted Patent: US 9,816,997 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/3053 Skin, nerves, brain
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/20 characterized by taxonomic origin
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 33/57496 involving intracellular compounds

Abstract

The present invention is related to the anti-SOX10 antibodies, kits, cocktails, and use of anti-SOX10 antibodies for detection of cancer.

Claims

1. An isolated preparation of antibodies which specifically bind to a SOX10 protein wherein said antibodies bind to an epitope in peptide SEQ ID NO:

3.

4. An isolated preparation of antibodies according to claim 1 wherein said antibodies are monoclonal.

5. An isolated preparation of antibodies according to claim 1 wherein said antibodies are polyclonal.

9. A method for producing a monoclonal antibody according to claim 1 comprising the steps of:

culturing said hybridoma which produces a monoclonal antibody capable of specifically recognizing SOX10; and

allowing said hybridoma to produce the monoclonal antibody.

12. An isolated preparation of antibodies according to claim 1 wherein said antibodies comprise a light chain variable region comprising the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO:

2 and a heavy chain variable region comprising the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO;

1.

13. An isolated preparation of antibodies according to claim 1 wherein said antibodies comprise a light chain variable region comprising the amino acid sequence of SEQ ID NO:

5 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

4.

22. An isolated preparation of antibodies according to claim 1 wherein said antibodies comprise a light chain variable region comprising the amino acid sequence selected from a group consisting SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 11, and any combination thereof.

23. An isolated preparation of antibodies according to claim 1 wherein said antibodies comprise a heavy chain variable region comprising the amino acid sequence selected from a group consisting SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, and any combination thereof.

29. A composition comprising at least two antibodies or fragments thereof, wherein at least one of said at least two antibodies or fragments thereof comprises said anti-SOX10 antibodies according to claim 1 which binds specifically to SOX10.

30. A composition according to claim 29 wherein said anti-SOX10 comprise the SOX10 antibodies.

43. A composition according to claim 29 wherein said composition comprises a primary antibody cocktail.

44. A composition according to claim 30 wherein said at least two antibodies or fragments thereof are derived from at least two different species.

45. A composition according to claim 44 wherein said at least two different species is selected from a group consisting of mouse, rabbit, goat, horse, chicken, human, and any combination thereof.

46. A composition according to claim 29 wherein said at least two antibodies or fragments thereof comprises a double-stain procedure.

47. A composition according to claim 29 wherein said at least two antibodies or fragments thereof are capable of providing different visualization results.

48. A composition according to claim 47 wherein said visualization results comprises color results.

49. A composition according to claim 29 wherein said at least one other of said at least two antibodies or fragments thereof binds specifically to a protein selected from a group consisting of Tyrosinase, MART-1, S100, and any combination thereof.

50. A composition according to claim 29 wherein said at least one other of said at least two antibodies or fragments thereof are selected from a group consisting of:

Tyrosinase antibody, MART-1 antibodies, Tyrosinase antibody [T311], MART-1 antibody [M2-7C10], MART-1 antibody [M2-9E3], and any combination thereof.

51. A composition according to claim 29 wherein said composition comprises a detection composition selected from a group consisting of melanoma detection, nevi detection, breast cancer detection, rhabdomyosarcoma detection, leiomyosarcoma detection, schwannomas detection, desmoplastic melanoma detection, spindle cell melanoma detection, and any combination thereof.

56. An antibody according to claim 4 wherein said monoclonal antibody is selected from a group consisting of a mouse monoclonal antibody, a rabbit monoclonal antibody, a goat monoclonal antibody, a horse monoclonal antibody, a chicken monoclonal antibody, a humanized monoclonal antibody, a chimeric antibody, and any combination thereof.

58. An antibody according to claim 5 wherein said polyclonal antibody is selected from a group consisting of rabbit polyclonal antibody, mouse polyclonal antibody, a goat polyclonal antibody, a horse polyclonal antibody, a chicken polyclonal antibody, a humanized polyclonal antibody, and any combination thereof.

61. An antibody according to claim 29 and further comprising a label attached to at least one of said at least two antibodies or fragments thereof.

63. An antibody according to claim 61 wherein said label is selected from a group consisting of a radioactive element, magnetic particles, radioisotope, fluorescent dye, enzyme, toxin, signal, stain, detection enzymes, horseradish peroxidase (HRP), alkaline phosphatase (AP), beta-galactosidase, chromogens, Fast Red, 3,3′

-diaminobenzidine, 3-amino-9-ethylcarbazole, 5-bromo-4-chloro-3-indolyl phosphate, 3,3′

,5,5′

-tetramethylbenzidine, 5-bromo-4-chloro-3-indolyl-β

-D-glucuronide, and any combination thereof.

68. Use of the antibody or fragment thereof according to claim 1 wherein said use is selected from a group consisting of:

use to detect cancer, use to diagnose or prognose cancer, use to predict outcome of treatment of cancer, use to assess efficacy of treatment of cancer, and use to predict recurrence of cancer.

77. A method for detecting a protein to which an antibody or fragment thereof of claim 1 binds to a biological sample, comprising the steps of contacting a biological sample with the antibody or fragment thereof;

and detecting the presence of the antibody or fragment thereof bound to the protein in the biological sample.

78. A method according to claim 77 wherein said biological sample comprises lung tissue, bladder tissue, breast tissue, and prostate tissue.

79. A method according to claim 77 wherein said biological sample is selected from a group consisting of a normal tissue, neoplastic tissue, bladder tissue, kidney tissue, ovarian tissue, thyroid tissue, endometrial tissue, renal tissue, tonsil tissue, pancreas tissue, colon tissue, lymph node tissue, neoplastic pancreatic tissue, stomach tissue, prostate tissue, lung tissue, skin tissue, and breast tissue.

80. A method according to claim 77 wherein said detecting said presence of the antibody or fragment thereof bound to the protein is performed on an automated staining device.

81. A method according to claim 77 wherein said detecting said presence of the antibody or fragment thereof bound to the protein corn rises detecting selected from a group consisting of:

manually detecting;

automatically detecting, and image analysis.

84. A method according to claim 77 wherein said detecting comprises a method selected from a group consisting of immunohistochemistry (IHC), IHC of FFPE, IHC of frozen-tissue sections, immunocytochemistry, and ELISA.

108. An immunoassay method for detecting SOX10 protein in an animal or human according to claim 77 and further comprising the steps of fixing or freezing said biological sample, and treating said fixed or frozen tissue to unmask epitopes to SOX10.

2. (canceled)

3. An isolated preparation of antibodies which specifically bind to an epitope in peptide SEQ ID NO:

3.

6. An antibody or fragment thereof produced by the hybridoma deposited with the American Type Culture Collection (ATCC) under ATCC Patent Deposit Designation No. PTA-120969.

7-8. -8. (canceled)

10-11. -11. (canceled)

14-21. -21. (canceled)

24-28. -28. (canceled)

31-42. -42. (canceled)

52-55. -55. (canceled)

57. (canceled)

59-60. -60. (canceled)

62. (canceled)

64-67. -67. (canceled)

69-76. -76. (canceled)

82-83. -83. (canceled)

85-107. -107. (canceled)

109-112. -112. (canceled)