Anti-human tenascin monoclonal antibody

Abstract

A novel anti-human tenascin ST2146 monoclonal antibody is described endowed with high affinity for the native antigen and high tumor selectivity. The cST2146 hybridoma is stably producing the antibody in high density culture conditions and is suitable for the industrial development of ST2146-based products. ST2146 exhibits properties exploitable for both therapeutic and diagnostic applications.

1 . Monoclonal antibody anti-human tenascin whose light and heavy chain variable region sequences are SEQ ID NO:1 and SEQ ID NO:2, respectively, and the proteolytic fragments thereof binding to an antigenic epitope within the EGF-like repeat of human tenascin C.  
     
     
         2 . The antibody and the fragments thereof according to  claim 1  further containing additional markers and/or diagnostic agents.  
     
     
         3 . Recombinant derivatives of the antibody and the fragments thereof of  claim 1  or  2 .  
     
     
         4 . Recombinant derivatives according to  claim 3 , wherein the murine constant region is replaced by the human counterpart or a biologically or pharmacologically active moiety.  
     
     
         5 . Recombinant derivatives according to  claim 4 , wherein said biologically active moiety is selected from the group consisting of: a member of the avidin family, a growth factor useful for stimulating tumor-directed immunological effectors.  
     
     
         6 . Recombinant derivatives according to  claim 4 , wherein said pharmacologically active moiety is selected from the group consisting of: a toxin, a superantigen, a cytokine or any other protein useful in enhancing the antitumor therapeutical efficacy.  
     
     
         7 . Conjugate derivatives of the antibody and the fragments thereof of  claim 1  or  2 .  
     
     
         8 . Conjugate according to  claim 7 , wherein a biologically or pharmacologically active moiety is linked to said antibody or the fragments thereof.  
     
     
         9 . Conjugate according to  claim 8 , wherein said biologically active moiety is selected from the group consisting of a growth factor useful for stimulating tumor-directed immunological effectors or said pharmacologically active moiety is selected from the group consisting of a toxin, a superantigen, a cytokine or any other protein useful in enhancing the antitumor therapeutical efficacy, antitumor drugs, radioisotopes.  
     
     
         10 . A biotinylated derivative of the antibody, or fragments or recombinant derivatives or conjugate thereof of any of claims  1 - 9 .  
     
     
         11 . The hybridoma cell line producing the antibody of  claim 1 , deposited with Advanced Biotechnology Center on Jan. 29, 2002 under the provision of Budapest Treaty, with the number PD02003.  
     
     
         12 . Process for the preparation of antibody of  claim 1 , comprising cultivating the hybridoma of  claim 11  and isolating said antibody.  
     
     
         13 . Use of the antibody or fragments, or recombinant derivatives or conjugates thereof of any of claims  1 - 9  or the biotinylated derivative of  claim 10  for a diagnostic means for the detection of diseases expressing tenascin.  
     
     
         14 . Use of  claim 13 , wherein said disease is a tumor.  
     
     
         15 . Use of  claim 14 , wherein said tumor is selected from the group consisting of gliomas, mammary, lung carcinomas, fibrosarcomas and squamous cell carcinomas.  
     
     
         16 . Use according to any of claims  13 - 15 , wherein said diagnostic means is used in in-vivo imaging techniques.  
     
     
         17 . Use of the antibody or fragments, or recombinant derivatives or conjugates thereof of any of claims  1 - 9  or the biotinylated derivative of  claim 10  for the preparation of a medicament for the treatment of diseases expressing tenascin.  
     
     
         18 . Use of  claim 17 , wherein said disease is a tumor.  
     
     
         19 . Use of  claim 18 , wherein said tumor is selected from the group consisting of gliomas, mammary, lung carcinomas, fibrosarcomas and squamous cell carcinomas.  
     
     
         20 . Use of any of claims  17 - 19 , wherein said medicament is in the form of a kit suitable for carrying out the three-step pre-targeting method.  
     
     
         21 . Therapeutic kit composed of 5 vials, whose first vial contains the the biotinylated derivative of  claim 10 , the second vial contains an avidin, the third vial contains biotinylated albumine, the fourth vial contains a streptavidin, the fifth vial contains radiolabelled biotin or biotin derivative.  
     
     
         22 . Therapeutic kit according to  claim 21 , wherein said vials are suitable for human injection.  
     
     
         23 . DNA encoding the antibody or and fragments thereof of  claim 1 .  
     
     
         24 . Specific CDRs and proteins comprising or containing CDRs.  
     
     
         25 . Vector containing DNA of  claim 23 .  
     
     
         26 . Host cells containing vectors of  claim 25 .  
     
     
         27 . Use of antibody or fragments, or recombinant derivatives or conjugates thereof of any of claims  1 - 9  in combination with a second tenascin-specific antibody in a sandwich assay for the production of a diagnostic kit for the determination of the level of circulating tenascin.  
     
     
         28 . Diagnostic kit comprising the antibody or fragments, or recombinant derivatives or conjugates thereof of any of claims  1 - 9 .  
     
     
         29 . A container, optionally containing multiple compartments, comprising the biotinylated antibody or fragments thereof of  claim 10 , buffers and reagents suitable for use in a therapeutic kit for a three-step pre-targeting method.  
     
     
         30 . The container according to  claim 29 , further comprising a separate tenascin-specific antibody preferentially directed towards the A-D fragment.  
     
     
         31 . The container according to  claim 29 , further comprising a separate tumor specific antibody.  
     
     
         32 . The antibody of  claim 1  or  claim 2  in combination with a second tenascin-specific antibody in a sandwich ELISA in vitro assay under conditions where said second antibody binds to a second antigenic epitope of tenascin, said in vitro ELISA assay useful for determining the level of circulating tenascin.  
     
     
         33 . Pharmaceutical composition comprising the antibody and/or a fragment and/or a recombinant derivative and/or a conjugate thereof of any of claims  1 - 9 , in admixture with at least one pharmaceutically acceptable excipient and/or vehicle.