Single-chain cd27-receptor agonist proteins

Abstract

Provided herein are specific CD27 receptor agonist proteins, nucleic acids encoding the same, and methods of treating a subject having a CD27L-associated disease or disorder. The CD27 receptor agonist proteins provided herein comprise three soluble CD27L domains an and Fc fragment. The CD27 receptor agonist proteins are substantially non-aggregating and suitable for therapeutic, diagnostic and/or research applications.

1 . A nucleic acid molecule encoding a cluster of differentiation 27 (CD27) receptor agonist protein, whereby said protein is a single-chain fusion polypeptide comprising:
 (i) a first soluble CD27L (CD27 ligand) domain,   (ii) a first peptide linker having 3 to 8 amino acids,   (iii) a second soluble CD27L domain,   (iv) a second peptide linker having 3 to 8 amino acids,   (v) a third soluble CD27L domain,   (vi) a hinge-linker selected from the group consisting of SEQ ID NOs: 16 and 19-24, and   (vii) an antibody Fc fragment consisting of the amino acid sequence of SEQ ID NO: 13 or amino acids 1-217 of SEQ ID NO: 13;   wherein each of the soluble CD27L domains (i), (iii), and (v) comprises amino acids Asp56-Pro193 of SEQ ID NO: 1.   
     
     
         2 . The nucleic acid molecule of  claim 1 , wherein each of the soluble CD27L domains (i), (iii), and (v) consists of amino acids Glu51-Pro193 or Asp56-Pro193 of SEQ ID NO: 1. 
     
     
         3 . The nucleic acid molecule of  claim 1 , wherein each of the soluble CD27L domains (i), (iii), and (v) consists of amino acids Glu51-Pro193 or Asp56-Pro193 of SEQ ID NO: 1, with one or more of the soluble CD27L domains (i), (iii), and (v) having a mutation at the amino acid position Glu51, Trp55, Asn63, Arg83, Arg122, Arg138, Arg144, His123, His124, His148, Asn170, Arg179, or Asp182 of SEQ ID NO: 1. 
     
     
         4 . The nucleic acid molecule of  claim 1 , in operative linkage with an expression control sequence. 
     
     
         5 . An expression vector comprising the nucleic acid molecule of  claim 4 . 
     
     
         6 . A cell or a non-human organism transformed or transfected with the nucleic acid molecule of  claim 4 . 
     
     
         7 . The nucleic acid molecule of  claim 1 , comprising SEQ ID NO: 37. 
     
     
         8 . The nucleic acid molecule of  claim 1 , comprising nucleotides 35 to 2146 of SEQ ID NO: 37. 
     
     
         9 . The nucleic acid molecule of  claim 1 , comprising nucleotides 95 to 2146 of SEQ ID NO: 37. 
     
     
         10 . An expression vector comprising the nucleic acid molecule of  claim 8 , in operative linkage with an expression control sequence. 
     
     
         11 . An expression vector comprising the nucleic acid molecule of  claim 9 , in operative linkage with an expression control sequence. 
     
     
         12 . A cell comprising the nucleic acid molecule of  claim 8 . 
     
     
         13 . A cell comprising the nucleic acid molecule of  claim 9 . 
     
     
         14 . The cell according to  claim 12 , which is a eukaryotic cell. 
     
     
         15 . The cell according to  claim 14 , wherein the cell is a mammalian cell. 
     
     
         16 . The cell according to  claim 15 , wherein the cell is a Chinese Hamster Ovary (CHO) cell. 
     
     
         17 . The cell according to  claim 13 , which is a eukaryotic cell. 
     
     
         18 . The cell according to  claim 17 , wherein the cell is a mammalian cell. 
     
     
         19 . The cell according to  claim 18 , wherein the cell is a Chinese Hamster Ovary (CHO) cell.