Anti-uroplakin II antibodies systems and methods

• US 9,429,577 B2
• Filed: 09/26/2013
• Issued: 08/30/2016
• Est. Priority Date: 09/27/2012
• Status: Active Grant

• Current Assignee: Biocare Medical LLC
• Original Assignee: Biocare Medical LLC
• Inventors: Qi, Weimin, Tacha, David
• Primary Examiner(s): Haq, Shafiqul
• Assistant Examiner(s): BELEI, CARMENCITA
• Application Number:
  US14/432,132
• Publication Number:
  US 20150247861A1
• Time in Patent Office: 1,069 Days
• US Class Current: 1/1
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/3038 Kidney, bladder
• CPC Class Codes: C07K 2317/20 characterized by taxonomic origin
• 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: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

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

Claims

1. An isolated antibody produced by the hybridoma deposited with the American Type Culture Collection (ATCC) under ATCC Patent Deposit Designation No. PTA-13181 or an isolated antigen binding fragment of the isolated antibody, wherein the antibody or antigen binding fragment thereof specifically binds Uroplakin II comprising SEQ ID NO:

4.

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

culturing said hybridoma deposited with the American Type Culture Collection (ATCC) under ATCC Patent Deposit Designation No. PTA-13181 which produces a monoclonal antibody capable of specifically recognizing Uroplakin II; and

allowing said hybridoma to produce the monoclonal antibody.

5. A composition comprising the isolated antibody or isolated antigen binding fragment thereof of claim 1 and at least one additional isolated antibody or isolated antigen binding fragment thereof, wherein the at least one additional isolated antibody or isolated antigen binding fragment thereof specifically binds to an antigen selected from the group consisting of GATA-3, p63, Uroplakin III, PAX8, NKX3.1, PSA and p40.

6. The composition according to claim 5 wherein said isolated antibody or isolated antigen binding fragment thereof and said at least one additional isolated antibody or isolated antigen binding fragment thereof each bind specifically to proteins selected from a group consisting of:

Uroplakin II and GATA-3;

Uroplakin II and p63;

Uroplakin II and Uroplakin III;

Uroplakin II and PAX8;

Uroplakin II and NKX3.1;

Uroplakin II and PSA;

Uroplakin II and Uroplakin III and GATA-3;

Uroplakin II and PAX8 and PSA;

Uroplakin II and PAX8 and NKX3.1; and

Uroplakin II and p40.

7. The isolated antibody or isolated antigen binding fragment thereof of claim 1, wherein the isolated antibody or isolated antigen is conjugated with a label.

8. An antibody according to claim 7 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.

9. A kit comprising:

the isolated antibody or isolated antigen binding fragment thereof of claim 1; and

an antibody detection reagent of said isolated antibody or isolated antigen binding fragment thereof of claim 1, when bound the Uroplakin II antigen.

10. A method for detecting Uroplakin II in a biological sample comprising the steps of:

(a) providing the kit of claim 9; and

(b) detecting binding of said isolated antibody or said isolated antigen binding fragment thereof with the Uroplakin II antigen with an immunoassay comprising;

(i) contacting the biological sample with the isolated antibody or isolated antigen binding fragment thereof produced by hybridoma deposited with the American Type Culture Collection (ATCC) under ATCC Patent Deposit Designation No. PTA-13181, wherein the antibody or antigen fragment thereof specifically binds Uroplakin II comprising SEQ ID NO;

4 to form a complex; and

(ii) detecting the complex with the antibody detecting reagent provided in the kit, wherein the immunoassay is selected from the group consisting of immunohistochemistry (IHC), IHC of FFPE, IHC of frozen-tissue sections, immunocytochemistry, and ELISA.

11. The method according to claim 10 wherein said biological sample comprises blood, urine, urothelial tissue, transitional cell tissue, and bladder tissue.

12. The method according to claim 10 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, stomach tissue, prostate tissue, lung tissue and breast tissue.

2. A hybridoma cell that is deposited at the American Type Culture Collection (ATCC) under ATCC Patent Deposit Designation No. PTA-13181.

3. The hybridoma cell according to claim 2 further comprising an antibody or antigen binding fragment thereof produced by said hybridoma cell.