CD27 ligand (CD27L) single-chain fusion polypeptides
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.
1. A single-chain fusion polypeptide comprising:
(i) a first soluble CD27 ligand (CD27L) cytokine domain,
(ii) a first peptide linker,
(iii) a second soluble CD27L cytokine domain,
(iv) a second peptide linker, and
(v) a third soluble CD27L cytokine domain,
wherein each of the soluble CD27L cytokine domains lacks a stalk region.
2. The polypeptide of claim 1 , wherein the first, the second, and the third soluble CD27L cytokine domains are, independently an N-terminally shortened domain of SEQ ID NO:7, and optionally consists of an amino acid mutation.
3. The polypeptide of claim 1 , wherein the first, the second, and the third soluble CD27L cytokine domains independently have the amino sequence of 51-193 or 56-193 of SEQ ID NO: 7.
4. The polypeptide of claim 1 , wherein the first and second peptide linkers are, independently, glycine/serine linkers.
5. The polypeptide of claim 4 , wherein the glycine/serine linkers comprise substituted asparagine residues.
6. The polypeptide of claim 1 , which additionally comprises an N-terminal signal peptide domain.
7. The polypeptide of claim 6 , wherein the N-terminal signal peptide domain comprises a protease cleavage site.
8. The polypeptide of claim 1 , which additionally comprises a further domain at the N-terminal and/or C-terminal end.
9. The polypeptide of claim 1 , which additionally comprises a further domain and a third peptide linker at the C-terminal end.
10. An isolated nucleic acid molecule encoding the fusion polypeptide of claim 1 .
11. An isolated host cell or a non-human organism transformed or transfected with the nucleic acid molecule of claim 10 .