SINGLE-CHAIN MULTIVALENT BINDING PROTEINS WITH EFFECTOR FUNCTION
Multivalent binding peptides, including bi-specific binding peptides, having immunoglobulin effector function are provided, along with encoding nucleic acids, vectors and host cells as well as methods for making such peptides and methods for using such peptides to treat or prevent a variety of diseases, disorders or conditions, as well as to ameliorate at least one symptom associated with such a disease, disorder or condition.
1 - 25 . (canceled)
26 . A polypeptide comprising, from amino-terminus to carboxy-terminus:
(a) a first binding domain;
(b) a constant sub-region comprising an immunoglobulin C H2 domain and a C H3 domain;
(c) a linker peptide comprising an amino acid sequence from a stalk region of a type II C-lectin protein; and
(e) a second binding domain.
27 . The polypeptide of claim 26 , wherein the first and/or second binding domain comprises variable regions from an immunoglobulin.
28 . The polypeptide of claim 26 , wherein the first and second binding domains specifically bind different target molecules located on the same cell.
29 . The polypeptide of claim 26 , wherein the first and second binding domains specifically bind different target molecules located on physically distinct cells.
30 . The polypeptide of claim 26 , wherein at least one binding domain specifically binds a cell-free molecular target.
31 . The polypeptide of claim 26 , wherein at least one binding domain is an scFv comprising a sequence selected from SEQ ID NO:2, 4, 6, 103, 105, 107, and 109.
32 . The polypeptide of claim 26 , wherein the type II C-lectin protein is selected from the group consisting of CD69, CD72, CD94, NKG2A, and NKG2D.
33 . The polypeptide of claim 26 , wherein the linker comprises a sequence selected from SEQ ID NOS: 373, 374, 375, 376, and 377.
34 . The polypeptide of claim 26 , wherein the linker is at least 5 amino acids in length.
35 . The polypeptide of claim 34 , wherein the linker is between 5 and 45 amino acids in length.
36 . The polypeptide of claim 26 , wherein the linker is resistant to proteolytic cleavage.
37 . The polypeptide of claim 27 , wherein the first and/or second binding domains comprise chimeric, humanized, or human immunoglobulin variable domains.
38 . The polypeptide of claim 26 , wherein the constant sub-region comprises IgG1 immunoglobulin C H2 and C H3 domains.
39 . The polypeptide of claim 26 , wherein the constant sub-region comprises C H2 and C H3 domains from a human immunoglobulin.
40 . The polypeptide of claim 26 , wherein the polypeptide does not comprise a C H1 domain.
41 . The polypeptide of claim 26 , wherein the constant sub-region further comprises an immunoglobulin hinge region.
42 . The polypeptide of claim 41 , wherein the hinge region is a hinge region selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgE, IgA2, synthetic hinge and the hinge-like C H2 domain of IgM.
43 . The polypeptide of claim 42 , wherein the hinge region is an IgG1 hinge region.
44 . The polypeptide of claim 43 , wherein the hinge region is a human IgG1 hinge region with a mutation at one or two cysteine residues.
45 . The polypeptide of claim 26 , wherein at least one binding domain specifically binds a target selected from a tumor antigen, a B-cell target, a TNF receptor superfamily member, a Hedgehog family member, a receptor tyrosine kinase, a proteoglycan-related molecule, a TGF-beta superfamily member, a Wnt-related molecule, a receptor ligand, a T-cell target, a Dendritic cell target, an NK cell target, a monocyte/macrophage cell target, a myeloid target, and an angiogenesis target.
46 . The polypeptide of claim 26 , wherein one of the two binding domains specifically binds a target selected from CD3, CD20, CD37, CD79b, CD80, CD86, IRTA1, IRTA2, IRTA3, IRTA4, IRTA5, TNFRI/TNFRSF1A, TNFRII/TNFRSF1B, Fas/TNFRSF6, TRAILR/TNFRSF10, RANK/TNFRSF11A, Osteoprotegerin/TNFRSF11B, TWEAKR/TNFRSF12, HVEM/TNFRSF14, GITR/TNFRSF18, TNF-α/TNFSF1A, TNF-β/TNFSF1B, TRAIL/TNFSF10, Fas Ligand/TNFSF6, TWEAK/TNFSF12, APRIL/TNFSF13, LIGHT/TNFSF14, GITRL/TNFSF18, FGFR, Flt-3, HGFR, IGF-IR, IGF-IIR, MSPR/Ron, PDGFRα, PDGFRβ, EGFR, ErbB2, ErbB3, VEGFR1/Flt-1, VEGFR2/Flk-1, VEGFR3/Flt-4, TGF-βRI/ALK-5, TGF-βRII, TGF-βRIIb, EGF, TGF-α, IGF-I, IGF-II, BMP, TGF-β, FGF, P1GF, PDGF-A, PDGF-B, PDGF-C, PDGF-D, VEGF, VEGF-B, VEGF-C, VEGF-D, IL-2R, IL-2Rβ, IL-4R, B7-H3, IL-6R, IL-10Rα, IL-10Rβ, IL-12Rβ1, IL-12Rβ2, IL-13Rα1, Osteopontin, PD-1, CTLA-4, IFN-γ R1, IFN-γ R2, Receptor for Advanced Glycation End products (RAGE), IL-13, IL-22R, IL-21, and IL-4.
47 . The polypeptide of claim 26 , wherein the polypeptide comprises a binding domain pair specifically recognizing a pair of targets selected from CD19/CD20, CD19/CD22, CD19/CL II, CD20/CD21, CD20/CD22, CD20/CD40, CD20/CD79a, CD20/CD79b, CD20/CD81, CD20/CL II, CD21/CD22, CD21/CD79b, CD21/CL II, CD22/CD23, CD22/CD30, CD22/CD37, CD22/CD40, CD22/CD70, CD22/CD72, CD22/CD79a, CD22/CD79b, CD22/CD80, CD22/CD86, CD22/CL II, CD23/CL II, CD30/CL II, CD37/CD79b, CD37/CL II, CD40/CD79b, CD40/CL II, CD70/CD79b, CD70/CL II, CD72/CD79b, CD72/CL II, CD79a/CD79b, CD79b/CD80, CD79b/CD81, CD79b/CD86, CD79b/CL II, CD80/CL II, and CD86/CL II.
48 . The polypeptide of claim 26 , wherein the immunoglobulin constant sub-region provides an effector function selected from the group consisting of antibody-dependent cell-mediated cytotoxicity, complement-dependent cytotoxicity, complement fixation, antibody-dependent cellular phagocytosis, binding to Fc receptors, and protein A binding.
49 . The polypeptide of claim 26 , wherein the polypeptide has an extended in vivo half-life as compared to a polypeptide lacking the constant sub-region.
50 . The polypeptide of claim 26 , wherein the polypeptide has an in vivo half-life of at least 28 hours in a human.
51 . The polypeptide of claim 26 , wherein the polypeptide is capable of forming dimers.
52 . A nucleic acid encoding the polypeptide of claim 26 .
53 . A vector comprising the nucleic acid of claim 52 .
54 . A host cell comprising the nucleic acid of claim 52 .
55 . A host cell comprising the vector of claim 53 .
56 . A composition comprising the polypeptide of claim 26 and a pharmaceutically acceptable carrier.