Prokaryotically produced antibodies and use thereof

Abstract

The present invention provides methods and compositions for improved expression and production of recombinant antibodies in prokaryotic expression systems. Particularly contemplated are prokaryotic expression and production of full length aglycosylated antibodies. The antibody products of the invention can be used in various aspects of biological research, diagnosis and medical treatment.

1 - 15 . (canceled)  
     
     
         16 . A process for producing a biologically active intact antibody in a prokaryotic host cell, said process comprising expressing in the host cell a polynucleotide comprising (1) a first promoter and a first cistron forming a first promoter-cistron pair and (2) a second promoter and a second cistron forming a second promoter-cistron pair, wherein the first cistron of said first promoter-cistron pair comprises a first translational initiation region (TIR-L) operably linked to a nucleic acid sequence encoding an immunoglobulin light chain and the second cistron of said second promoter-cistron pair comprises a second translational initiation region (TIR-H) operably linked to a nucleic acid sequence encoding an immunoglobulin heavy chain, wherein upon expression of said polynucleotide, secreted light chain and heavy chain are folded and assembled to form a biologically active intact antibody; and recovering said intact antibody.  
     
     
         17 - 27 . (canceled)  
     
     
         28 . An glycosylated full length antibody produced by a process according to  claim 16 .  
     
     
         29 . The aglycosylated full length antibody of  claim 28 , wherein the immunoglobulin is a multispecific antibody.  
     
     
         30 . The aglycosylated full length antibody of  claim 28 , which is a non-human antibody.  
     
     
         31 . The aglycosylated full length antibody of  claim 30 , wherein the non-human antibody is humanized.  
     
     
         32 . The aglycosylated full length antibody of  claim 28 , which is a human antibody.  
     
     
         33 . An immunoconjugate comprising the aglycosylated full length antibody of  claim 28  conjugated with a cytotoxic agent.  
     
     
         34 . The immunoconjugate of  claim 33 , wherein the cytotoxic agent is selected from the group consisting of a radioactive isotope, a chemotherapeutic agent and a toxin.  
     
     
         35 . The immunoconjugate of  claim 34 , wherein the toxin is selected from the group consisting of calicheamicin, maytansine and trichothene.  
     
     
         36 . A composition comprising the aglycosylated full length antibody of  claim 28  and a carrier.  
     
     
         37 . The composition of  claim 36 , wherein the carrier is pharmaceutically acceptable.  
     
     
         38 . A composition comprising the immunoconjugate of  claim 33  and a carrier.  
     
     
         39 . The composition of  claim 38 , wherein he carrier is pharmaceutically acceptable.  
     
     
         40 . An article of manufacture comprising a) a container and a composition contained therein, wherein the composition comprises an aglycosylated full length of  claim 28;  and b) instruction for using said composition.  
     
     
         41 . An article of manufacture comprising a) a container and a composition contained therein, wherein the composition comprises an immunoconjugate according to  claim 33;  and b) instruction for using said composition.