Multivariable Antigens Complexed with Targeting Humanized Monoclonal Antibody

Abstract

The present invention includes compositions and methods for designing, making and using modular recombinant antibodies or fragments thereof with one half of a cohesin-dockerin pair that permits the rapid assembly of multivariant antigen conjugates.

1 . A modular rAb carrier comprising an antigen-specific binding domain linked to one or more antigen carrier domains comprising one half of a cohesin-dockerin binding pair. 
     
     
         2 .- 10 . (canceled) 
     
     
         11 . A vaccine comprising a modular rAb carrier comprising an antigen specific domain linked to one or more domains comprising one half of the cohesin-dockerin binding pair bound to a complementary half of the cohesin-dockerin binding pair bound to an antigen. 
     
     
         12 . The vaccine of  claim 11 , wherein the antigen specific domain is specific for an immune cell surface protein selected from MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40,CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor or other receptor relatively specifically expressed by antigen presenting cells. 
     
     
         13 . The vaccine of  claim 11 , wherein the antigen comprises a bacterial, viral, fungal, protozoan or cancer protein. 
     
     
         14 . The vaccine of  claim 11 , wherein the modular rAb carrier is further defined:
 an rAb.Doc:Coh.antigen;   an rAb.Coh:Doc.antigen;   an rAb.(Coh) x :(Doc.antigen) x ;   an rAb.(Doc) x :(Coh.antigen) x ;   an rAb.(Coh.Doc) x :(Doc.antigen 1 )(Coh.antigen 2 ); or an rAb.(Coh) x (Doc) x :(Doc.antigen 1 ) x (Coh.antigen 2 ) x ;   wherein x is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.   
     
     
         15 . An isolated nucleic acid comprising a coding segment for an target specific domain and one or more domains and one half of a cohesin-dockerin binding pair. 
     
     
         16 . The nucleic acid of  claim 15 , wherein the target is an antigen and the specific domain encodes at least a portion of an antibody. 
     
     
         17 . The nucleic acid of  claim 15 , wherein the one or more domains encodes one or more cohesin domains, one or more dockerin domains or a combination of one or more cohesin and dockerin domains. 
     
     
         18 . The nucleic acid of  claim 15 , wherein the target specific domain comprises an rAb is further defined as: an rAb.Doc; an rAb.Coh; an rAb.(Coh) x ; an rAb.(Doc) x ; an rAb.(Coh.Doc) x ; or an rAb.(Coh) x (Doc) x ; wherein x is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. 
     
     
         19 . A vector comprising a nucleic acid encoding an antigen specific domain and one or more domains that comprise one half of a cohesin-dockerin binding pair, a one half of a cohesin-dockerin binding pair with a protein molecule to be carried and combinations thereof. 
     
     
         20 . The vector of  claim 19 , wherein the one half of a cohesin-dockerin binding pair, a one half of a cohesin-dockerin binding pair with a protein molecule to be carried and combinations thereof are under the control of the same promoter, different promoters, transcribed in-line, transcribed in opposite directions. 
     
     
         21 . A host cell comprising a vector comprising a nucleic acid encoding an antigen specific domain and one or more domains and one half of a cohesin-dockerin binding pair. 
     
     
         22 . A method of making a modular rAb carrier comprising:
 combining an antigen specific domain linked to one or more domains comprising one half of a cohesin-dockerin binding pair.   
     
     
         23 . The method of  claim 22 , wherein the rAb is further defined as: an rAb.Doc; an rAb.Coh; an rAb.(Coh) x ; an rAb.(Doc) x ; an rAb.(Coh.Doc) x ; or an rAb.(Coh) x (Doc) x ; wherein x is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. 
     
     
         24 . The method of  claim 22 , wherein the rAb is complexed with a complementary half of a cohesion:dokerin pair bound to an antigen and is selected from:
 an rAb.Doc:Coh.antigen;   an rAb.Coh:Doc.antigen;   an rAb.(Coh) x :(Doc.antigen) x ;   an rAb.(Doc) x :(Coh.antigen) x ;   an rAb.(Coh.Doc) x : (Doc.antigen 1 )(Coh.antigen 2 ); or   an rAb.(Coh) x (Doc) x :(Doc.antigen 1 ) x (Coh.antigen 2 ) x ;   wherein x is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.   
     
     
         25 . An immunotoxin comprising an rAb.Doc:Coh.toxin self-assembled conjugate, wherein the rAb is specific for a cell target. 
     
     
         26 . The immunotoxin of  claim 25 , wherein the toxin is selected from wherein the toxin is selected from the group consisting of a radioactive isotope, metal, enzyme, botulin, tetanus, ricin, cholera, diphtheria, aflatoxins, perfringens toxin, mycotoxins, shigatoxin, staphylococcal enterotoxin B, T2, seguitoxin, saxitoxin, abrin, cyanoginosin, alphatoxin, tetrodotoxin, aconotoxin, snake venom and spider venom. 
     
     
         27 . The immunotoxin of  claim 25 , wherein the cell target comprises a cancer cell selected from hematological cancers, leukemias, lymphomas, neurological tumors, astrocytomas or glioblastomas, melanoma, breast cancer, lung cancer, head and neck cancer, gastrointestinal tumors such as gastric or colon cancer, liver cancer, pancreatic cancer, genitourinary tumors such cervix, uterus, ovarian cancer, vaginal cancer, testicular cancer, prostate cancer or penile cancer, bone tumors, vascular tumors, or cancers of the lip, nasopharynx, pharynx and oral cavity, esophagus, rectum, gall bladder, biliary tree, larynx, lung and bronchus, bladder, kidney, brain and other parts of the nervous system, thyroid, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma and leukemia. 
     
     
         28 . The immunotoxin of  claim 25 , wherein the cell target comprises a pathogen selected from a bacteria, a protozoan, a helminth, a virally-infected cell or a fungus. 
     
     
         29 . A method for protein purification, comprising:
 separating a cohesin or dockerin fusion protein by interacting the fusion protein with a rAb that is conjugated to the complementary cohesin or dockerin bound to a substrate.   
     
     
         30 . The method of  claim 29 , further comprising the step of administering the protein in a therapeutic application comprising transplantation, autoimmune disease, infectious disease or cancer. 
     
     
         31 . The use of the cohesin as a fusion partner for toxins for conferring beneficial biochemical properties favoring ready purification of active cohesin.toxin fusion protein. 
     
     
         32 . The use of anti-DC rAb.Doc to target DC for therapeutic applications where ablating DC. 
     
     
         33 . An anti-DC-SIGN/L antibody provided in an amount that is sufficient to enhance the survival of dendritic cells, wherein the antibody matures and activates the dendritic cells for immunization. 
     
     
         34 . The antibody of  claim 33 , wherein the antibody is targeted in vivo to dendritic cells as an adjuvant in vaccines. 
     
     
         35 . (canceled) 
     
     
         36 . A bivalent and multivalent (rAb.Doc:Coh.cytokine), (rAb.Coh:Doc.cytokine) or (cytokine 1 .Coh:cytokine 2  Doc) self-assembled conjugates as therapeutic, cell proliferation or maturing agents. 
     
     
         37 . A method for making modular rAb comprising:
 screening one or more multivalent rAb and/or rAb.cytokine and/or cytokine cytokine combinations that are capable of specifically binding to a target cell and delivering the cytokine such that it exerts its effect on the target cell.   
     
     
         38 . The method of  claim 37 , wherein the cytokine comprises interleukins, transforming growth factors (TGFs), fibroblast growth factors (FGFs), platelet derived growth factors (PDGFs), epidermal growth factors (EGFs), connective tissue activated peptides (CTAPs), osteogenic factors, and biologically active analogs, fragments, and derivatives of such growth factors, B/T-cell differentiation factors, B/T-cell growth factors, mitogenic cytokines, chemotactic cytokines, colony stimulating factors, angiogenesis factors, IFN-α, IFN-β, IFN-γ, IL1, IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL9, IL10, IL11, IL12, IL13, IL14, IL15, IL16, IL17, IL18, etc., leptin, myostatin, macrophage stimulating protein, platelet-derived growth factor, TNF-α, TNF-β, NGF, CD40L, CD137L/4-1BBL, human lymphotoxin-β, G-CSF, M-CSF, GM-CSF, PDGF, IL-1α, IL1-β, IP-10, PF4, GRO, 9E3, erythropoietin, endostatin, angiostatin, VEGF, transforming growth factor (TGF) supergene family include the beta transforming growth factors (for example TGF-β1, TGF-β2, TGF-β3); bone morphogenetic proteins (for example, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9); heparin-binding growth factors (fibroblast growth factor (FGF), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF)); Inhibins (for example, Inhibin A, Inhibin B); growth differentiating factors (for example, GDF-1); and Activins (for example, Activin A, Activin B, Activin AB).