Fusion polypeptides capable of activating receptors
A fusion polypeptide comprising (A) x -M-(A′) y , wherein A and A′ are each polypeptides capable of binding a target receptor. The fusion polypeptides of the invention form multimeric proteins which activate the target receptor. A and A′ may be each be an antibody or fragment derived from an antibody specific for a target receptor, such as the same or different scFv fragments, and/or a ligand or ligand fragment or derivative capable of binding the target protein, M is a multimerizing component, and X and Y are independently a number between 1-10.
1. A dimer formed from two fusion polypeptides of the form (A)-M-(A′), wherein:
component A comprises an antibody fragment specific to a target receptor,
component A′ comprises either
i) an antibody fragment specific to the target receptor; or
ii) a ligand or ligand fragment specific to the target receptor, and
component M is a multimerizing component comprising an Fc domain of IgG,
the dimer being characterized in that, when contacted with a system comprising the target receptor, it binds to and clusters four or more molecules of the target receptor, wherein the target receptor is selected from the group consisting of an EGF receptor, Eph receptor, EPO receptor, FGF receptor, G-CSF receptor, GM-CSF receptor, a VEGF receptor, a PDGF receptor, TGFβ receptor, a Tie receptor and a Trk receptor.
2. The dimer of claim 1 , wherein A and A′ are different antibody fragments.
3. The dimer of claim 1 , wherein A and A′ are the same antibody fragment.
4. The dimer of claim 1 , wherein A′ is a ligand.
5. The dimer of claim 1 , wherein A′ is a ligand fragment.
6. The dimer of claim 1 , wherein M comprises an Fc domain of human IgG1.
7. A pharmaceutical composition comprising the dimer of claim 1 and a pharmaceutically acceptable carrier.
8. A kit comprising at least one container, therein containing the dimer of claim 1 .
9. A nucleic acid molecule encoding a fusion polypeptide which binds to a target receptor selected from the group consisting of an EGF receptor, an Eph receptor, an EPO receptor, an FGF receptor, a G-CSF receptor, a GM-CSF receptor, a VEGF receptor, a PDGF receptor, a TGFβ receptor, a Tie receptor, and a Trk receptor, the fusion polypeptide having the form (A)-M-(A′), wherein:
component A comprises an antibody fragment specific to said target receptor,
component A′ comprises either
i) an antibody fragment specific to said target receptor; or
ii) a ligand or ligand fragment specific to said target receptor, and component M is a multimerizing component comprising an Fc domain of IgG.
10. A vector comprising the nucleic acid molecule of claim 9 .
11. A host cell for the production of a fusion polypeptide comprising the vector of claim 10 .
12. The host cell of claim 11 , wherein the host cell is selected from the group consisting of bacterial cells, yeast cells, insect cells and mammalian cells.
13. The host cell of claim 12 , wherein the host cell is selected from the group consisting of E. coli, Pichia pastoris, Spodoptera frugiperda , COS, CHO, 293, BHK and a NSO cell.
14. The host cell of claim 13 , wherein the host cell is a CHO cell.
15. A method of producing a fusion polypeptide comprising culturing the host cell of claim 11 under conditions sufficient to express the fusion polypeptide from the host cell, and recovering the fusion polypeptide.