Multivalent heteromultimer scaffold design and constructs
Provided herein are multifunctional heteromer proteins. In specific embodiments is a heteromultimer that comprises: at least two monomeric proteins, wherein each monomeric protein comprises at least one cargo polypeptide, attached to a transporter polypeptide, such that said monomeric proteins associate to form the heteromultimer. These therapeutically novel molecules comprise monomers that function as scaffolds for the conjugation or fusion of therapeutic molecular entities resulting in the creation of bispecific or multivalent molecular species.
1. A heteromultimer comprising a first monomeric protein that comprises (i) a first transporter polypeptide comprising a first segment of albumin; and (ii) at least one cargo polypeptide and a second monomeric protein that comprises (iii) a second transporter polypeptide comprising a second segment of albumin and (iv) at least one cargo polypeptide; wherein said first segment of albumin and said second segment of albumin are derived from an albumin by segmentation of the albumin to remove a loop, said first transporter polypeptide is different from said second transporter polypeptide, and said transporter polypeptides self-assemble to form a quasi-native albumin structure.
2. The heteromultimer of claim 1 , wherein said heteromultimer is a heterodimer.
3. The heteromultimer of claim 1 wherein said albumin is human serum albumin having a sequence of SEQ ID NO: 1 or analog or variant thereof.
4. The heteromultimer of claim 1 , wherein said first transporter polypeptide has a sequence comprising SEQ ID NO:2 or an analog or variant thereof, and wherein said second transporter polypeptide has a sequence comprising SEQ ID NO:3 or an analog or variant thereof.
5. The heteromultimer of claim 1 , wherein said first transporter polypeptide has a sequence comprising SEQ ID NO:8 or an analog or variant thereof, and wherein said second transporter polypeptide has a sequence comprising SEQ ID NO:10or an analog or variant thereof.
6. The heteromultimer of claim 1 , wherein at least one transporter polypeptide is an allo-albumin derivative.
7. The heteromultimer according to claim 1 , wherein the transporter polypeptides are derived from the same albumin.
8. The heteromultimer of claim 7 , wherein said first transporter polypeptide and said second transporter polypeptide are derived from the same alloalbumin.
9. The heteromultimer according to claim 1 , wherein the transporter polypeptides are derived from different albumins.
10. The heteromultimer of claim 9 , wherein one of said first transporter polypeptide and said second transporter polypeptide is derived from human serum albumin and the other is derived from an alloalbumin.
11. The heteromultimer of claim 1 , wherein at least one cargo polypeptide is an antibody, or a fragment or variant thereof.
12. The heteromultimer of claim 1 wherein the at least one cargo polypeptide of said first monomeric protein binds a target antigen, and the at least one cargo polypeptide of said second monomeric protein comprises a toxin moiety.
13. The heteromultimer of claim 1 , wherein said at least one cargo polypeptide binds a target antigen, wherein said target antigen is at least one of alpha-chain (CD25) of IL-2R, Amyloid beta, anti-EpCAM, anti-CD3, CD16, CD20, CD22, CD23, CD3, CD4, CD52, CD80, CTLA-4, EGFR, EpCAM, F protein of RSV, G250, glycoprotein IIB/IIIa R, HER2, HSP90, IgE antibody, IL-12, IL-23, IL-1 beta, IL-5, IL-6, RANKL, TNF alpha, TNFR, VEGF-A, glucagon receptor, GLP receptor, and LDL receptor.
14. The heteromultimer of claim 1 wherein at least one cargo polypeptide is an enzyme, hormone, therapeutic polypeptide, antigen, chemotoxin, radiotoxin, cytokine or a variant or fragment thereof.
15. The heteromultimer of claim 1 , wherein said first monomeric protein and said second monomeric protein comprise the same cargo polypeptide.
16. The heteromultimer of claim 1 , wherein the cargo polypeptide is attached to the transporter polypeptide by chemical conjugation, native ligation, chemical ligation, a disulfide bond, or direct fusion or fusion via a linker.
17. The heteromultimer of claim 16 , wherein the linker is a GGSG linker or a (GGSG) 4 GG linker.
18. The heteromultimer of claim 1 , wherein the heteromultimer binds to FcRn.
19. The heteromultimer of claim 1 , wherein the first segment of albumin and the second segment of albumin form a complementary pair of transporter polypeptides.
20. The heteromultimer of claim 1 , wherein said first transporter polypeptide and said second transporter polypeptide are derived from a mammalian albumin.
21. The heteromultimer of claim 1 , wherein said first transporter polypeptide and said second transporter polypeptide are derived from a non-mammalian albumin.
22. A host cell comprising nucleic acid encoding the heteromultimer of claim 1 .
23. The host cell of claim 22 , wherein the nucleic acid encoding the first monomeric-protein and the nucleic acid encoding the second monomeric protein are present in a single vector.
24. The host cell of claim 22 , wherein the nucleic acid encoding the first monomeric protein and the nucleic acid encoding the second monomeric protein are present in separate vectors.