Monclonal antibodies, hybridomas and methods for use

• US 20120122118A1
• Filed: 11/07/2011
• Published: 05/17/2012
• Est. Priority Date: 01/25/2008
• Status: Abandoned Application

• Current Assignee: HansaBioMed OU, Primm SRL
• Original Assignee: HansaBioMed OU, Primm SRL
• Inventors: Lozupone, Francesco, Fais, Stefano, Chiesi, Antonio, Zarovni, Natasa, Sarmientos, Paolo, Pontillo, Angela Simona
• Application Number:
  US13/290,186
• Publication Number:
  US 20120122118A1
• Time in Patent Office: Days
• US Class Current: 435/7.5
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/30 from tumour cells
• 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: G01N 33/574 for cancer

Abstract

The present invention provides monoclonal antibodies to TM9SF-proteins and hybridoma cell lines that produce the monoclonal antibodies to TM9SF4. This invention also provides a method for determining the level of TM9SF4-protein in biological fluid samples, tissue samples and in microvesicles such as exosomes, comprising kit for determining the level of TM9SF4 protein in human exosomes and other microvesicles, in tissue samples, and in biological fluids.

Claims

1. A hybridoma cell line selected from the group consisting of hybridoma cell lines 1A4-A3, 1A4-A8, 1A4-F2, 1A4-G1, 5C1-B4, 5C1-C5, 5C1-D4, and 5C1-G6.

2. An isolated monoclonal antibody or a fragment thereof, wherein the antibody is produced by the hybridoma cell line of claim 1.

3. The monoclonal antibody or the fragment of claim 2, wherein the antibody or the fragment is conjugated to a detectable label.

4. The monoclonal antibody or the fragment of claim 2, wherein the antibody or the fragment is conjugated to a drug.

5. The monoclonal antibody of claim 4, wherein the drug is a medicinal compound approved for cancer therapy.

6. The monoclonal antibody of claim 4, wherein the drug is a medicinal compound approved for treatment of a TM9SF4 protein related disease condition.

7. An isolated mouse anti-TMSF4 antibody or an antigen binding fragment thereof, wherein the antibody binds to SEQ ID NO:

17 of TMSF4-protein and wherein the antibody is produced by a hybridoma cell line of claim 1.

11. A kit to detect TM9SF4-protein, said kit comprising clones of monoclonal antibodies or fragments thereof of claim 2 and a microplate or microtiter plate.

12. The kit of claim 11, wherein the kit further comprises purified TM9SF4 N-terminal domain fragment corresponding to SEQ ID NO:

17 and optionally isolated TM9SF4 protein according to SEQ ID NO;

2, rabbit anti-TM9SF4 polyclonal antibodies, a biotinylated anti-mouse IgG, horseradish peroxidase (HRP)-conjugated streptavidin, a HRP-conjugated anti mouse secondary antibody, a biotinylated anti-rabbit IgG or a HRP-conjugated anti rabbit secondary antibody, a phosphate buffer solution, a washing buffer solution, a diluent for a biological sample suspected to contain TM9SF4, a chromogenic or luminometric substrate, and a stop solution.

13. The kit of claim 11, wherein the kit is for an Enzyme-Linked Immunosorbent Assay (ELISA) and wherein the monoclonal antibodies are bound to the microplate or the microtiter plate.

8. An isolated mouse anti-TMSF1 antibody or antigen binding fragment thereof, wherein the antibody binds to SEQ ID NO:

19 of TMSF1-protein.

9. An isolated mouse anti-TMSF-2 antibody or antigen binding fragment thereof, wherein the antibody binds to SEQ ID NO:

20 of TMSF2-protein.

10. An isolated mouse anti-TMSF-3 antibody or antigen binding fragment thereof, wherein the antibody binds to SEQ ID NO:

21 of TMSF3-protein.

14. A pharmaceutical composition comprising therapeutically effective amount of an antibody of claim or an antigen binding fragment thereof.