Single-chain fusion polypeptide comprising trail

Abstract

The present invention refers to single-chain fusion proteins comprising three soluble TNF superfamily (TNFSF) cytokine domains and nucleic acid molecules encoding these fusion proteins. The fusion proteins are substantially non-aggregating and suitable for therapeutic, diagnostic and/or research applications.

What is claimed is: 
     
         1 . A single-chain fusion polypeptide comprising: (i) a first soluble TNF-related apoptosis-inducing ligand (TRAIL) cytokine domain, (ii) a first peptide linker, (iii) a second soluble TRAIL cytokine domain, (iv) a second peptide linker, and (v) a third soluble TRAIL cytokine domain. 
     
     
         2 . The polypeptide of  claim 1 , wherein the first and second peptide linkers are independently glycine/serine linkers. 
     
     
         3 . The polypeptide of  claim 2 , wherein the glycine/serine linkers are each independently 3-15 amino acids in length. 
     
     
         4 . The polypeptide of  claim 3 , wherein at least one of the soluble TRAIL domains ends with amino acid G281 of SEQ ID NO: 10 and comprises one or more mutations at positions R130, G160, H168, R170, H177, Y189, R191, Q193, E195, N199, K201, Y213, T214, S215, H264, I266, D267 or D269 of SEQ ID NO: 10. 
     
     
         5 . The polypeptide of  claim 4 , wherein the polypeptide additionally comprises a further domain at the N-terminal and/or C-terminal end. 
     
     
         6 . The polypeptide of  claim 5 , wherein the further domain is a Fc fragment domain. 
     
     
         7 . A pharmaceutical composition comprising the fusion polypeptide of  claim 6  and a pharmaceutically acceptable carrier, diluent and/or adjuvant. 
     
     
         8 . An isolated nucleic acid molecule encoding the fusion polypeptide of  claim 6 . 
     
     
         9 . A vector comprising the isolated nucleic acid molecule of  claim 8 . 
     
     
         10 . An isolated host cell or a non-human organism transformed or transfected with the vector of  claim 9 .