Novel oprf/i fusion proteins, their preparation and use

Abstract

The present invention relates to a novel trimeric OprF/I fusion protein comprising a portion of the Pseudomonas aeruginosa outer membrane protein F which is fused with its carboxy terminal end to a portion of the amino terminal end of the Pseudomonas aeruginosa out membrane protein I, wherein said portion of the Pseudomonas aeruginosa outer membrane protein F comprises the amino acids 190-342 of SEQ ID NO: 1 and wherein said portion of the Pseudomonas aeruginosa outer membrane protein I comprises the amino acids 21-83 of SEQ ID NO: 2, and further to a novel Opr F/I fusion protein which contains a disulphide bond pattern, preferably selected from the group consisting of (a) Cys18-Cys27-bond, (b) Cys18-Cys27-bond and Cys33-Cys47-bond, and (c) Cys18-Cys47 and Cys27-Cys33-bond, and to immunogenic variants thereof having at least 85% identity to the amino acid sequence of SEQ ID NO: 3. The present invention also relates to a novel method for producing said OprF/I fusion proteins and to their use for the preparation of a pharmaceutical composition and for the preparation of antibodies or antibody derivatives which specifically bind said novel OprF/I fusion proteins.

1 .- 12 . (canceled) 
     
     
         13 . An antibody or antibody derivative which specifically binds a protein complex comprising three OprF/I fusion proteins of SEQ ID NO: 4 or an immunogenic variant thereof having at least 85% identity to the amino acid sequence of SEQ ID NO: 4. 
     
     
         14 . An antibody or antibody derivative which selectively binds a protein complex comprising three OprF/I fusion proteins of SEQ ID NO: 4 or an immunogenic variant thereof having at least 85% identity to the amino acid sequence of SEQ ID NO: 4. 
     
     
         15 . A pharmaceutical composition comprising the antibody or antibody derivative according to  claim 13  and one or more pharmaceutically acceptable excipients. 
     
     
         16 . A pharmaceutical composition comprising the antibody or antibody derivative according to  claim 14  and one or more pharmaceutically acceptable excipients. 
     
     
         17 . The antibody or antibody derivative of  claim 13 , wherein the OprF/I fusion proteins are selected from the group consisting of
 (a) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond (SEQ ID NO: 9),   (b) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond and Cys33-Cys47-bond (SEQ ID NO: 10),   (c) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys47-bond and Cys27-Cys33-bond (SEQ ID NO: 11), and   immunogenic variants thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in (a), (b) or (c).   
     
     
         18 . The antibody or antibody derivative of  claim 17 , wherein the sum of a) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond (SEQ ID NO: 9), b) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond and Cys33-Cys47-bond (SEQ ID NO: 10), and c) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys47-bond and Cys27-Cys33-bond (SEQ ID NO: 11) is equal or greater than 75%. 
     
     
         19 . The antibody or antibody derivative of  claim 14 , wherein the OprF/I fusion proteins are selected from the group consisting of
 (a) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond (SEQ ID NO: 9),   (b) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond and Cys33-Cys47-bond (SEQ ID NO: 10),   (c) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys47-bond and Cys27-Cys33-bond (SEQ ID NO: 11), and   immunogenic variants thereof having at least 85% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in either (a), (b) or (c).   
     
     
         20 . The antibody or antibody derivative of  claim 19 , wherein the sum of a) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond (SEQ ID NO: 9), b) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond and Cys33-Cys47-bond (SEQ ID NO: 10), and c) the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys47-bond and Cys27-Cys33-bond (SEQ ID NO: 11) is equal or greater than 75%. 
     
     
         21 . The antibody or antibody derivative of  claim 13 , wherein the OprF/I fusion proteins are the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond (SEQ ID NO: 9) or an immunogenic variant thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in SEQ ID NO: 9. 
     
     
         22 . The antibody or antibody derivative of  claim 13 , wherein the OprF/I fusion proteins are the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond and Cys33-Cys47-bond (SEQ ID NO: 10) or an immunogenic variant thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in SEQ ID NO: 10. 
     
     
         23 . The antibody or antibody derivative of  claim 13 , wherein the OprF/I fusion proteins are the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys47-bond and Cys27-Cys33-bond (SEQ ID NO: 11) or an immunogenic variant thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in SEQ ID NO: 11. 
     
     
         24 . The antibody or antibody derivative of  claim 14 , wherein the OprF/I fusion proteins are the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond (SEQ ID NO: 9) or an immunogenic variant thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in SEQ ID NO: 9. 
     
     
         25 . The antibody or antibody derivative of  claim 14 , wherein the OprF/I fusion proteins are the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys27-bond and Cys33-Cys47-bond (SEQ ID NO: 10) or an immunogenic variant thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in SEQ ID NO: 10. 
     
     
         26 . The antibody or antibody derivative of  claim 14 , wherein the OprF/I fusion proteins are the OprF/I fusion protein of SEQ ID NO: 4 with a Cys18-Cys47-bond and Cys27-Cys33-bond (SEQ ID NO: 11) or an immunogenic variant thereof having at least 85%, preferably 90%, in particular 95% identity to the amino acid sequence of SEQ ID NO: 4, and the same disulphide bond pattern as specified in SEQ ID NO: 11.