Methods and compositions for the production of monoclonal antibodies

Abstract

The present invention comprises compositions and methods for making monoclonal antibodies. The present invention further comprises vectors that replicate the immune system components, particularly an antigen-presenting cell (APC) element of the immune synapse. Additionally, the present invention may further comprise synthetic T-cells.

1 . A vector composition, comprising a synthetic APC, wherein the synthetic APC comprises a colloidal metal, an antigen and a component specific immunostimulating agent.  
     
     
         2 . The vector composition of  claim 1 , wherein the component specific immunostimulating agent comprises antigens, colloidal metals, adjuvants, receptor molecules, nucleic acids, immunogenic proteins, accessory cytokine/immunostimulators, pharmaceuticals, chemotherapy agents or carriers.  
     
     
         3 . The vector composition of  claim 1 , wherein the component specific immunostimulating agent comprises IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-11, IL-12, IL-13, lipid A, phospholipase A2, endotoxins,  staphylococcal  enterotoxin B, Type I interferon, Type II interferon, Tumor Necrosis Factor (TNF-α), Flt-3 ligand, Transforming Growth Factor (TGF-β), lymphotoxin, migration inhibition factor, Granulocyte-Macrophage colony stimulating factor (CSF), monocyte-macrophage CSF, Granulocyte CSF, Vascular epithelial growth factor (VEGF), angiogenin, Transforming Growth Factor (TGF-α), heat shock proteins, carbohydrate moieties of blood groups, Rh factors, fibroblast growth factor, inflammatory and immune regulatory proteins, nucleotides, DNA, RNA, mRNA, sense, antisense, polynucleotides, cancer cell specific antigens, mutant p53, tyrosinase, mucines, autoimmune antigens, immunotherapy drugs, angiogenic or anti-angiogenic drugs.  
     
     
         4 . The component specific immunostimulating agent of  claim 3 , wherein the cancer cell specific antigen is MAGE, BAGE or MART.  
     
     
         5 . The component specific immunostimulating agent of  claim 3 , wherein the mucine is MUC-1, PSA or TSH.  
     
     
         6 . The vector composition of  claim 1 , wherein the colloidal metal comprises colloidal gold colloidal silver, colloidal iron, colloidal aluminum, or colloidal platinum.  
     
     
         7 . The vector composition of  claim 1 , further comprising a pharmaceutically-acceptable component comprising excipients, buffers or carriers.  
     
     
         8 . The vector composition of  claim 1 , further comprising an adjuvant, wherein the adjuvant comprises liposomes, emulsions, microspheres, biodegradable polymers and polystyrene, alum, heat killed  M. butyricum  and  M. tuberculosis, Pertussis  toxin and  Tetanus  toxin, or LPS and  Staphylococcal  enterotoxin B.  
     
     
         9 . The vector composition of  claim 1 , wherein the vector further comprises a co-stimulatory protein.  
     
     
         10 . The vector composition of  claim 1 , wherein the vector further comprises a major histocompatibility complex.  
     
     
         11 . The vector composition of  claim 1 , wherein the vector further comprises a germinal center.  
     
     
         12 . The vector composition of  claim 1 , wherein the vector further comprises a structural protein.  
     
     
         13 . The vector composition of  claim 1 , further comprising targeting molecules, integrating molecules, PEG, derivatized PEG, poly-L-lysine or derivatized poly-L-lysine, biotin-avidin, bifunctional PEG, or bifunctional polymers.  
     
     
         14 . The vector composition of  claim 13 , wherein the bifunctional PEG comprises thiol and biotin.  
     
     
         15 . The vector composition of  claim 13 , wherein the poly-L-lysine comprises thiol and biotin.  
     
     
         16 . The vector composition of  claim 1 , wherein the synthetic APC is lyophilized and stored for a period of time prior to administration.  
     
     
         17 . A method for the production of monoclonal antibodies in vitro, comprising: 
 a) admixing or binding an antigen, a component specific immunostimulating agent with a colloidal metal to form a synthetic APC; and    b) stimulating immune cells from a human in vitro to activate the immune cells to produce a primary response to a synthetic APC; wherein the synthetic APC comprises an antigen, a component specific immunostimulating agent, and a colloidal metal; and    c) immortalizing the activated immune cells; and    d) selecting a monoclonal antibody producing cell.    
     
     
         18 . The method of  claim 17 , further comprising a major histocompatibility complex.  
     
     
         19 . The method of  claim 17 , further comprising a germinal center.  
     
     
         20 . An antigen-specific, human-specific monoclonal antibody comprised entirely of human protein, produced by a process of: 
 a) stimulating immune cells from a human in vitro to activate the immune cells to produce a primary response to a synthetic APC; wherein the synthetic APC comprises an antigen, a component specific immunostimulating agent, and a colloidal metal; and    b) immortalizing the activated immune cells; and    c) selecting a monoclonal antibody producing cell.    
     
     
         21 . A method for affecting an immune response, comprising, administering a composition comprising at least one colloidal metal particle, an antigen, and at least one component-specific immunostimulating agent.  
     
     
         22 . A method for enhancing vaccine efficacy comprising administering a composition comprising at least one colloidal metal particle, an antigen, and at least one component-specific immunostimulating agent.  
     
     
         23 . A method of treating disease and immune related dysfunctions and pathologies, comprising, administering to a human or animal a therapeutically effective amount of a vector composition comprising a synthetic APC, wherein the synthetic APC comprises at least one colloidal metal, an antigen and an component specific immunostimulating agent.  
     
     
         24 . The method of  claim 23 , wherein the colloidal metal particles are of different sizes.  
     
     
         25 . The method of  claim 24 , wherein administering the component specific immunostimulating agent, the antigen, and the colloidal metal particles of different sizes comprises simultaneously administering the component specific immunostimulating agent, the antigen and the colloidal metal particles of different sizes as a single dose.  
     
     
         26 . The method of  claim 23 , wherein the method is used to stimulate or suppress immune components in the treatment of disease, wherein the disease is cancer, solid tumor cancer, Crohn's disease, breast cancer, Psoriasis, inflammatory bowel disease, adult respiratory distress syndrome, allergies, rhinitis, eczema, urticaria, anaphylaxis, transplant rejection, rheumatic diseases, systemic lupus erthymatosus, rheumatoid arthritis, seronegative spondyloarthritides, Sjogren's syndrome, systemic sclerosis, polymyositis, dermatomyositis, Type I Diabetes Mellitus, Acquired Immune Deficiency Syndrome, Hashimoto's thyroiditis, Graves' disease, Addison's disease, polyendocrine autoimmune disease, hepatitis, sclerosing cholangitis, primary biliary cirrhosis, pernicious anemia, coeliac disease, antibody-mediated nephritis, glomerulonephritis, Wegener's granulomatosis, microscopic polyarteritis, polyarteritis nodosa, pemphigus, dermatitis herpetiformis, psoriasis, vitiligo, multiple sclerosis, encephalomyelitis, Guillain-Barre syndrome, Myasthenia Gravis, Lambert-Eaton syndrome, sclera, episclera, uveitis, chronic mucocutaneous candidiasis, Bruton's syndrome, transient hypogammaglobulinemia of infancy, myeloma, X-linked hyper IgM syndrome, Wiskott-Aldrich syndrome, ataxia telangiectasia, autoimmune hemolytic anemia, autoimmune thrombocytopenia, autoimrune neutropenia, Waldenstrom's macroglobulinemia, amyloidosis, chronic lymphocytic leukemia, or non-Hodgkin's lymphoma.  
     
     
         27 . The method of  claim 23 , further comprising a major histocompatibility complex.  
     
     
         28 . A multi-particle self assembling synthetic APC, comprising at least two colloidal metals, an antigen and at least one component specific immunostimulating agent, wherein the multi-particle synthetic APC is assembled using scaffolding molecules, and wherein the scaffolding molecules comprises biotinylated spacer arms, alkane linkers, protein, polyethylene glycol, derivatized PEG, multi-arm PEG or thiolated poly-L-lysine.  
     
     
         29 . The multi-particle self-assembling synthetic APC of  claim 28 , wherein the colloidal metal particles are of different sizes.  
     
     
         30 . The multi-particle self-assembling synthetic APC of  claim 28 , further comprising targeting molecules and integrating molecules.  
     
     
         31 . The method of  claim 28 , further comprising a major histocompatibility complex.