Dual variable domain immunoglobulins and uses thereof

Abstract

Engineered multivalent and multispecific binding proteins, methods of making, and their uses in the prevention, diagnosis, and/or treatment of disease are provided.

1 - 60 . (canceled) 
     
     
         61 . A binding protein comprising first and second polypeptide chains, each polypeptide chain independently comprising VD1-(X1)n-VD2-C-(X2)n, wherein
 VD1 is a first variable domain;   VD2 is a second variable domain;   C is a constant domain;   X1 is a linker;   X2 is an Fc region; and   n is 0 or 1,   wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site, wherein:   (a) X1 between the first and second variable domains is ASTKGP (SEQ ID NO: 21) on the first polypeptide chain and TVAAPSVFIFPP (SEQ ID NO: 14) on the second polypeptide chain;   (b) X1 between the first and second variable domains is ASTKGPSVFPLAP (SEQ ID NO: 22) on the first polypeptide chain and TVAAP (SEQ ID NO: 13) on the second polypeptide chain; or   (c) X1 between the first and second variable domains is ASTKGPSVFPLAP (SEQ ID NO: 22) on the first polypeptide chain and TVAAPSVFIFPP (SEQ ID NO: 14) on the second polypeptide chain;   and wherein the binding protein is capable of binding:   1) TNF and SOST, wherein   a. the variable domains that form a functional target binding site for TNF comprise:   CDRs 1-3 from SEQ ID NO: 36 and CDRs 1-3 from SEQ ID NO: 37,   CDRs 1-3 from SEQ ID NO: 38 and CDRs 1-3 from SEQ ID NO: 39,   CDRs 1-3 from SEQ ID NO: 40 and CDRs 1-3 from SEQ ID NO: 41,   CDRs 1-3 from SEQ ID NO: 44 and CDRs 1-3 from SEQ ID NO: 45, or   CDRs 1-3 from SEQ ID NO: 46 and CDRs 1-3 from SEQ ID NO: 47; and   b. the variable domains that form a functional target binding site for SOST comprise CDRs 1-3 from SEQ ID NO: 32 and CDRs 1-3 from SEQ ID NO: 33;   2) TNF and PGE2, wherein   a. the variable domains that form a functional target binding site for TNF comprise:   CDRs 1-3 from SEQ ID NO: 36 and CDRs 1-3 from SEQ ID NO: 37,   CDRs 1-3 from SEQ ID NO: 38 and CDRs 1-3 from SEQ ID NO: 39,   CDRs 1-3 from SEQ ID NO: 40 and CDRs 1-3 from SEQ ID NO: 41,   CDRs 1-3 from SEQ ID NO: 44 and CDRs 1-3 from SEQ ID NO: 45, or   CDRs 1-3 from SEQ ID NO: 46 and CDRs 1-3 from SEQ ID NO: 47; and   b. the variable domains that form a functional target binding site for PGE2 comprise CDRs 1-3 from SEQ ID NO: 34 and CDRs 1-3 from SEQ ID NO: 35;   or   3) TNF and LPA, wherein   a. the variable domains that form a functional target binding site for TNF comprise:   CDRs 1-3 from SEQ ID NO: 36 and CDRs 1-3 from SEQ ID NO: 37,   CDRs 1-3 from SEQ ID NO: 38 and CDRs 1-3 from SEQ ID NO: 39,   CDRs 1-3 from SEQ ID NO: 40 and CDRs 1-3 from SEQ ID NO: 41,   CDRs 1-3 from SEQ ID NO: 44 and CDRs 1-3 from SEQ ID NO: 45, or   CDRs 1-3 from SEQ ID NO: 46 and CDRs 1-3 from SEQ ID NO: 47; and   b. the variable domains that form a functional target binding site for LPA comprise CDRs 1-3 from SEQ ID NO: 42 and CDRs 1-3 from SEQ ID NO: 43.   
     
     
         62 . The binding protein of  claim 61 , wherein the binding protein is capable of binding
 1) TNF and SOST, wherein
 a. the variable domains that form a functional target binding site for TNF comprise a sequence selected from the group consisting of SEQ ID NO: 36-41 and 44-47; and 
 b. the variable domains that form a functional target binding site for SOST comprise SEQ ID NO: 32 and/or SEQ ID NO: 33; 
   2) TNF and PGE2, wherein
 a. the variable domains that form a functional target binding site for TNF comprise a sequence selected from the group consisting of SEQ ID NO: 36-41 and 44-47; and 
 b. the variable domains that form a functional target binding site for PGE2 comprise SEQ ID NO: 34 and/or SEQ ID NO: 35; 
 or 
   3) TNF and LPA, wherein
 a. the variable domains that form a functional target binding site for TNF comprise a sequence selected from the group consisting of SEQ ID NO: 36-41 and 44-47; and 
 b. the variable domains that form a functional target binding site for LPA comprise SEQ ID NO: 42 and/or SEQ ID NO: 43. 
   
     
     
         63 . The binding protein of  claim 61 , wherein the binding protein comprises two first polypeptide chains and two second polypeptide chains, and four functional target binding sites. 
     
     
         64 . The binding protein of  claim 61 , wherein the first polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein
 VD1 is a first heavy chain variable domain;   VD2 is a second heavy chain variable domain;   C is a heavy chain constant domain;   X1 is a linker;   X2 is an Fc region;   n is 0 or 1; and   wherein the second polypeptide chains comprises VD1-(X1)n-VD2-C-(X2)n, wherein   VD1 is a first light chain variable domain;   VD2 is a second light chain variable domain;   C is a light chain constant domain;   X1 is a linker;   n is 0 or 1 for (X1)n; and   n is 0 for (X2)n.   
     
     
         65 . The binding protein of  claim 61 , wherein the binding protein is capable of binding
 1) TNF and SOST, wherein the binding protein comprises any one of:   DVD1453 (comprising SEQ ID NOs: 72 and 73); DVD1454 (comprising SEQ ID NOs: 74 and 75); DVD1455 (comprising SEQ ID NOs: 76 and 77); DVD1456 (comprising SEQ ID NOs: 78 and 79); DVD1457 (comprising SEQ ID NOs: 80 and 81); DVD1458 (comprising SEQ ID NOs: 82 and 83); DVD1475 (comprising SEQ ID NOs: 84 and 85); DVD1476 (comprising SEQ ID NOs: 86 and 87); DVD1483 (comprising SEQ ID NOs: 88 and 89); DVD1484 (comprising SEQ ID NOs: 90 and 91); DVD1495 (comprising SEQ ID NOs: 156 and 157); DVD1496 (comprising SEQ ID NOs: 158 and 159); DVD1497 (comprising SEQ ID NOs: 160 and 161); DVD1498 (comprising SEQ ID NOs: 162 and 163); DVD1499 (comprising SEQ ID NOs: 164 and 165); DVD1500 (comprising SEQ ID NOs: 166 and 167); DVD1517 (comprising SEQ ID NOs: 168 and 169); DVD1518 (comprising SEQ ID NOs: 170 and 171); DVD1525 (comprising SEQ ID NOs: 172 and 173); DVD1526 (comprising SEQ ID NOs: 174 and 175); DVD1537 (comprising SEQ ID NOs: 240 and 241); DVD1538 (comprising SEQ ID NOs: 242 and 243); DVD1539 (comprising SEQ ID NOs: 244 and 245); DVD1540 (comprising SEQ ID NOs: 246 and 247); DVD1541 (comprising SEQ ID NOs: 248 and 249); DVD1542 (comprising SEQ ID NOs: 250 and 251); DVD1559 (comprising SEQ ID NOs: 252 and 253); DVD1560 (comprising SEQ ID NOs: 254 and 255); DVD1567 (comprising SEQ ID NOs: 256 and 257); and DVD1568 (comprising SEQ ID NOs: 258 and 259);   2) TNF and PGE2, wherein the binding protein comprises any one of:   DVD1447 (comprising SEQ ID NOs: 92 and 93); DVD1448 (comprising SEQ ID NOs: 94 and 95); DVD1449 (comprising SEQ ID NOs: 96 and 97); DVD1450 (comprising SEQ ID NOs: 98 and 99); DVD1451 (comprising SEQ ID NOs: 100 and 101); DVD1452 (comprising SEQ ID NOs: 102 and 103); DVD1473 (comprising SEQ ID NOs: 104 and 105); DVD1474 (comprising SEQ ID NOs: 106 and 107); DVD1481 (comprising SEQ ID NOs: 108 and 109); DVD1482 (comprising SEQ ID NOs: 110 and 111); DVD1489 (comprising SEQ ID NOs: 176 and 177); DVD1490 (comprising SEQ ID NOs: 178 and 179); DVD1491 (comprising SEQ ID NOs: 180 and 181); DVD1492 (comprising SEQ ID NOs: 182 and 183); DVD1493 (comprising SEQ ID NOs: 184 and 185); DVD1494 (comprising SEQ ID NOs: 186 and 187); DVD1515 (comprising SEQ ID NOs: 188 and 189); DVD1516 (comprising SEQ ID NOs: 190 and 191); DVD1523 (comprising SEQ ID NOs: 192 and 193); DVD1524 (comprising SEQ ID NOs: 194 and 195); DVD1531 (comprising SEQ ID NOs: 260 and 261); DVD1532 (comprising SEQ ID NOs: 262 and 263); DVD1533 (comprising SEQ ID NOs: 264 and 265); DVD1534 (comprising SEQ ID NOs: 266 and 267); DVD1535 (comprising SEQ ID NOs: 268 and 269); DVD1536 (comprising SEQ ID NOs: 270 and 271); DVD1557 (comprising SEQ ID NOs: 272 and 273); DVD1558 (comprising SEQ ID NOs: 274 and 275); DVD1565 (comprising SEQ ID NOs: 276 and 277); and DVD1566 (comprising SEQ ID NOs: 278 and 279);   or   3) TNF and LPA, wherein the binding protein comprises any one of:   DVD1465 (comprising SEQ ID NOs: 112 and 113); DVD1466 (comprising SEQ ID NOs: 114 and 115); DVD1467 (comprising SEQ ID NOs: 116 and 117); DVD1468 (comprising SEQ ID NOs: 118 and 119); DVD1469 (comprising SEQ ID NOs: 120 and 121); DVD1470 (comprising SEQ ID NOs: 122 and 123); DVD1471 (comprising SEQ ID NOs: 124 and 125); DVD1472 (comprising SEQ ID NOs: 126 and 127); DVD1479 (comprising SEQ ID NOs: 128 and 129); DVD1480 (comprising SEQ ID NOs: 130 and 131); DVD1487 (comprising SEQ ID NOs: 132 and 133); DVD1488 (comprising SEQ ID NOs: 134 and 135); DVD1507 (comprising SEQ ID NOs: 196 and 197); DVD1508 (comprising SEQ ID NOs: 198 and 199); DVD1509 (comprising SEQ ID NOs: 200 and 201); DVD1510 (comprising SEQ ID NOs: 202 and 203); DVD1511 (comprising SEQ ID NOs: 204 and 205); DVD1512 (comprising SEQ ID NOs: 206 and 207); DVD1513 (comprising SEQ ID NOs: 208 and 209); DVD1514 (comprising SEQ ID NOs: 210 and 211); DVD1521 (comprising SEQ ID NOs: 212 and 213); DVD1522 (comprising SEQ ID NOs: 214 and 215); DVD1529 (comprising SEQ ID NOs: 216 and 217); DVD1530 (comprising SEQ ID NOs: 218 and 219); DVD1549 (comprising SEQ ID NOs: 280 and 281); DVD1550 (comprising SEQ ID NOs: 282 and 283); DVD1551 (comprising SEQ ID NOs: 284 and 285); DVD1552 (comprising SEQ ID NOs: 286 and 287); DVD1553 (comprising SEQ ID NOs: 288 and 289); DVD1554 (comprising SEQ ID NOs: 290 and 291); DVD1555 (comprising SEQ ID NOs: 292 and 293); DVD1556 (comprising SEQ ID NOs: 294 and 295); DVD1563 (comprising SEQ ID NOs: 296 and 297); DVD1564 (comprising SEQ ID NOs: 298 and 299); DVD1571 (comprising SEQ ID NOs: 300 and 301); and DVD1572 (comprising SEQ ID NOs: 302 and 303).   
     
     
         66 . The binding protein of  claim 61 , wherein:
 1) The binding protein is capable of binding TNF and SOST, and is capable of binding to TNF with a dissociation constant (K D ) of at most about 2.10×10 −10  M as measured by surface plasmon resonance, and/or is capable of binding to SOST with a dissociation constant (K D ) of at most about 2.20×10 −8  M as measured by surface plasmon resonance;   2) The binding protein is capable of binding TNF and PGE2, and is capable of binding to TNF with a dissociation constant (K D ) of at most about 9.20×10 −11  M as measured by surface plasmon resonance;   or   3) The binding protein is capable of binding TNF and LPA, and is capable of binding to TNF with a dissociation constant (K D ) of at most about 5.5×10 −10  M as measured by surface plasmon resonance.   
     
     
         67 . The binding protein of  claim 61 , wherein the Fc region is a variant sequence Fc region. 
     
     
         68 . The binding protein of  claim 61 , wherein the Fc region is from an IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or an IgD. 
     
     
         69 . The binding protein of  claim 61 , wherein the binding protein has a dissociation constant (K D ) to at least one target of at most about 10 −7  M; at most about 10 −8  M; at most about 10 −9  M; at most about 10 −10  M; at most about 10 −11  M; at most about 10 −12  M; or at most 10 −13 M, as measured by surface plasmon resonance. 
     
     
         70 . The binding protein of  claim 61 , wherein the binding protein is a crystallized binding protein. 
     
     
         71 . A binding protein conjugate comprising the binding protein of  claim 61 , the binding protein conjugate further comprising an immunoadhension molecule, an imaging agent, a therapeutic agent, or a cytotoxic agent. 
     
     
         72 . The binding protein conjugate of  claim 71 , wherein the imaging agent is a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, or biotin. 
     
     
         73 . The binding protein conjugate of  claim 72 , wherein the radiolabel is  3 H,  14 C,  35 S,  90 Y,  99 Tc,  111 In,  125 I,  131 I,  177 Lu,  166 Ho, or  153 Sm. 
     
     
         74 . The binding protein conjugate of  claim 71 , wherein the therapeutic agent or cytotoxic agent is an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, a toxin, or an apoptotic agent. 
     
     
         75 . An isolated nucleic acid encoding the binding protein of  claim 61 . 
     
     
         76 . A vector comprising the isolated nucleic acid of  claim 75 . 
     
     
         77 . An isolated host cell comprising the vector of  claim 76 . 
     
     
         78 . The host cell of  claim 77 , wherein the host cell is selected from the group consisting of a prokaryotic cell, an  Escherichia coli  cell, a eukaryotic cell, an animal cell, a plant cell, a fungal cell, a mammalian cell, a yeast cell, an avian cell, an insect cell, a CHO cell, a COS cell, an  Saccharomyces cerevisiae  cell, and an Sf9 cell. 
     
     
         79 . A method of producing a binding protein, the method comprising the step of culturing the host cell of  claim 77  in culture medium under conditions sufficient to produce the binding protein. 
     
     
         80 . A pharmaceutical composition comprising the binding protein of  claim 61  and a pharmaceutically acceptable carrier. 
     
     
         81 . The pharmaceutical composition of  claim 80 , further comprising at least one additional therapeutic agent selected from the group consisting of an imaging agent, a cytotoxic agent, an angiogenesis inhibitor, a kinase inhibitor, a co-stimulation molecule blocker, an adhesion molecule blocker, an anti-cytokine antibody or functional fragment thereof, methotrexate, cyclosporin, rapamycin, FK506, a detectable label or reporter, a TNF antagonist, an antirheumatic, a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteroid, an anabolic steroid, an erythropoietin, an immunization, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist. 
     
     
         82 . A method of treating a subject for a disease or a disorder, the method comprising the step of administering to the subject the binding protein of  claim 61  such that treatment is achieved. 
     
     
         83 . The method of  claim 82 , wherein the disease or disorder is selected from the group consisting of rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, septic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic lupus erythematosus, Crohn's disease, ulcerative colitis, inflammatory bowel disease, insulin dependent diabetes mellitus, thyroiditis, asthma, allergic diseases, psoriasis, dermatitis scleroderma, graft versus host disease, organ transplant rejection, acute or chronic immune disease associated with organ transplantation, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki's disease, Grave's disease, nephrotic syndrome, chronic fatigue syndrome, Wegener's granulomatosis, Henoch-Schoenlein purpurea, microscopic vasculitis of the kidneys, chronic active hepatitis, uveitis, septic shock, toxic shock syndrome, sepsis syndrome, cachexia, infectious diseases, parasitic diseases, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington's chorea, Parkinson's disease, Alzheimer's disease, stroke, primary biliary cirrhosis, hemolytic anemia, malignancies, heart failure, Addison's disease, sporadic, polyglandular deficiency type I and polyglandular deficiency type II, Schmidt's syndrome, adult (acute) respiratory distress syndrome, alopecia, alopecia areata, arthropathy, Reiter's disease, psoriatic arthropathy, ulcerative colitic arthropathy, enteropathic synovitis, chlamydia, yersinia and salmonella associated arthropathy, atheromatous disease/arteriosclerosis, atopic allergy, autoimmune bullous disease, pemphigus vulgaris, pemphigus foliaceus, pemphigoid, linear IgA disease, autoimmune haemolytic anaemia, Coombs positive haemolytic anaemia, acquired pernicious anaemia, juvenile pernicious anaemia, myalgic encephalitis/Royal Free Disease, chronic mucocutaneous candidiasis, giant cell arteritis, primary sclerosing hepatitis, cryptogenic autoimmune hepatitis, acquired immunodeficiency related diseases, hepatitis B, hepatitis C, common varied immunodeficiency (common variable hypogammaglobulinaemia), dilated cardiomyopathy, female infertility, ovarian failure, premature ovarian failure, fibrotic lung disease, cryptogenic fibrosing alveolitis, post-inflammatory interstitial lung disease, interstitial pneumonitis, connective tissue disease associated interstitial lung disease, mixed connective tissue disease associated lung disease, systemic sclerosis associated interstitial lung disease, rheumatoid arthritis associated interstitial lung disease, systemic lupus erythematosus associated lung disease, dermatomyositis/polymyositis associated lung disease, Sjögren's disease associated lung disease, ankylosing spondylitis associated lung disease, vasculitic diffuse lung disease, haemosiderosis associated lung disease, drug-induced interstitial lung disease, fibrosis, radiation fibrosis, bronchiolitis obliterans, chronic eosinophilic pneumonia, lymphocytic infiltrative lung disease, postinfectious interstitial lung disease, gouty arthritis, autoimmune hepatitis, type-1 autoimmune hepatitis (classical autoimmune or lupoid hepatitis), type-2 autoimmune hepatitis (anti-LKM antibody hepatitis), autoimmune mediated hypoglycaemia, type B insulin resistance with acanthosis nigricans, hypoparathyroidism, acute immune disease associated with organ transplantation, chronic immune disease associated with organ transplantation, osteoarthrosis, primary sclerosing cholangitis, psoriasis type 1, psoriasis type 2, idiopathic leucopaenia, autoimmune neutropaenia, renal disease NOS, glomerulonephritides, microscopic vasulitis of the kidneys, lyme disease, discoid lupus erythematosus, male infertility idiopathic or NOS, sperm autoimmunity, multiple sclerosis (all subtypes), sympathetic ophthalmia, pulmonary hypertension secondary to connective tissue disease, Goodpasture's syndrome, pulmonary manifestation of polyarteritis nodosa, acute rheumatic fever, rheumatoid spondylitis, Still's disease, systemic sclerosis, Sjörgren's syndrome, Takayasu's disease/arteritis, autoimmune thrombocytopaenia, idiopathic thrombocytopaenia, autoimmune thyroid disease, hyperthyroidism, goitrous autoimmune hypothyroidism (Hashimoto's disease), atrophic autoimmune hypothyroidism, primary myxoedema, phacogenic uveitis, primary vasculitis, vitiligo acute liver disease, chronic liver diseases, alcoholic cirrhosis, alcohol-induced liver injury, choleosatatis, idiosyncratic liver disease, drug-induced hepatitis, non-alcoholic steatohepatitis, allergy and asthma, group B streptococci (GBS) infection, mental disorders, depression, schizophrenia, Th2 Type and Th1 Type mediated diseases, acute and chronic pain, different forms of pain, cancers, lung cancer, breast cancer, stomach cancer, bladder cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer, rectal cancer, hematopoietic malignancies, leukemia, lymphoma, Abetalipoprotemia, acrocyanosis, acute and chronic parasitic or infectious processes, acute leukemia, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute or chronic bacterial infection, acute pancreatitis, acute renal failure, adenocarcinomas, aerial ectopic beats, AIDS dementia complex, alcohol-induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergic rhinitis, allograft rejection, alpha-1-antitrypsin deficiency, amyotrophic lateral sclerosis, anemia, angina pectoris, anterior horn cell degeneration, anti-CD3 therapy, antiphospholipid syndrome, anti-receptor hypersensitivity reactions, aortic and peripheral aneuryisms, aortic dissection, arterial hypertension, arteriosclerosis, arteriovenous fistula, ataxia, atrial fibrillation (sustained or paroxysmal), atrial flutter, atrioventricular block, B cell lymphoma, bone graft rejection, bone marrow transplant (BMT) rejection, bundle branch block, Burkitt's lymphoma, burns, cardiac arrhythmias, cardiac stun syndrome, cardiac tumors, cardiomyopathy, cardiopulmonary bypass inflammation response, cartilage transplant rejection, cerebellar cortical degenerations, cerebellar disorders, chaotic or multifocal atrial tachycardia, chemotherapy associated disorders, chronic myelocytic leukemia (CML), chronic alcoholism, chronic inflammatory pathologies, chronic lymphocytic leukemia (CLL), chronic obstructive pulmonary disease (COPD), chronic salicylate intoxication, colorectal carcinoma, congestive heart failure, conjunctivitis, contact dermatitis, cor pulmonale, coronary artery disease, Creutzfeldt-Jakob disease, culture negative sepsis, cystic fibrosis, cytokine therapy associated disorders, dementia pugilistica, demyelinating diseases, dengue hemorrhagic fever, dermatitis, dermatologic conditions, diabetes, diabetes mellitus, diabetic ateriosclerotic disease, diffuse Lewy body disease, dilated congestive cardiomyopathy, disorders of the basal ganglia, Down's syndrome in middle age, drug-induced movement disorders induced by drugs which block CNS dopamine receptors, drug sensitivity, eczema, encephalomyelitis, endocarditis, endocrinopathy, epiglottitis, epstein-barr virus infection, erythromelalgia, extrapyramidal and cerebellar disorders, familial hematophagocytic lymphohistiocytosis, fetal thymus implant rejection, Friedreich's ataxia, functional peripheral arterial disorders, fungal sepsis, gas gangrene, gastric ulcer, glomerular nephritis, graft rejection of any organ or tissue, gram negative sepsis, gram positive sepsis, granulomas due to intracellular organisms, hairy cell leukemia, Hallervorden-Spatz disease, Hashimoto's thyroiditis, hay fever, heart transplant rejection, hemachromatosis, hemodialysis, hemolytic uremic syndrome/thrombolytic thrombocytopenic purpura, hemorrhage, hepatitis A, His bundle arrythmias, HIV infection/HIV neuropathy, Hodgkin's disease, hyperkinetic movement disorders, hypersensitity reactions, hypersensitivity pneumonitis, hypertension, hypokinetic movement disorders, hypothalamic-pituitary-adrenal axis evaluation, idiopathic Addison's disease, idiopathic pulmonary fibrosis, antibody mediated cytotoxicity, Asthenia, infantile spinal muscular atrophy, inflammation of the aorta, influenza a, ionizing radiation exposure, iridocyclitis/uveitis/optic neuritis, ischemia-reperfusion injury, ischemic stroke, juvenile rheumatoid arthritis, juvenile spinal muscular atrophy, Kaposi's sarcoma, kidney transplant rejection, legionella, leishmaniasis, leprosy, lesions of the corticospinal system, lipedema, liver transplant rejection, lymphederma, malaria, malignant lymphoma, malignant histiocytosis, malignant melanoma, meningitis, meningococcemia, metabolic/idiopathic, migraine headache, mitochondrial multisystem disorder, mixed connective tissue disease, monoclonal gammopathy, multiple myeloma, multiple systems degenerations (Mencel Dejerine-Thomas Shi-Drager and Machado-Joseph), mycobacterium avium intracellulare, mycobacterium tuberculosis, myelodyplastic syndrome, myocardial infarction, myocardial ischemic disorders, nasopharyngeal carcinoma, neonatal chronic lung disease, nephritis, nephrosis, neurodegenerative diseases, neurogenic muscular atrophies, neutropenic fever, non-hodgkins lymphoma, occlusion of the abdominal aorta and its branches, occulsive arterial disorders, okt3 therapy, orchitis/epidydimitis, orchitis/vasectomy reversal procedures, organomegaly, osteoporosis, pancreas transplant rejection, pancreatic carcinoma, paraneoplastic syndrome/hypercalcemia of malignancy, parathyroid transplant rejection, pelvic inflammatory disease, perennial rhinitis, pericardial disease, peripheral atherlosclerotic disease, peripheral vascular disorders, peritonitis, pernicious anemia, pneumocystis carinii pneumonia, pneumonia, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes syndrome), post perfusion syndrome, post pump syndrome, post-MI cardiotomy syndrome, preeclampsia, progressive supranucleo palsy, primary pulmonary hypertension, radiation therapy, Raynaud's phenomenon and disease, Raynoud's disease, Refsum's disease, regular narrow QRS tachycardia, renovascular hypertension, reperfusion injury, restrictive cardiomyopathy, sarcomas, scleroderma, senile chorea, senile dementia of Lewy body type, seronegative arthropathies, shock, sickle cell anemia, skin allograft rejection, skin changes syndrome, small bowel transplant rejection, solid tumors, specific arrythmias, spinal ataxia, spinocerebellar degenerations, streptococcal myositis, structural lesions of the cerebellum, subacute sclerosing panencephalitis, syncope, syphilis of the cardiovascular system, systemic anaphalaxis, systemic inflammatory response syndrome, systemic onset juvenile rheumatoid arthritis, T-cell or FAB ALL telangiectasia, thromboangitis obliterans, thrombocytopenia, toxicity, transplants, trauma/hemorrhage, type III hypersensitivity reactions, type IV hypersensitivity, unstable angina, uremia, urosepsis, valvular heart diseases, varicose veins, vasculitis, venous diseases, venous thrombosis, ventricular fibrillation, viral and fungal infections, vital encephalitis/aseptic meningitis, vital-associated hemaphagocytic syndrome, Wernicke-Korsakoff syndrome, Wilson's disease, xenograft rejection of any organ or tissue, acute coronary syndromes, acute idiopathic polyneuritis, acute inflammatory demyelinating polyradiculoneuropathy, acute ischemia, adult Still's disease, anaphylaxis, anti-phospholipid antibody syndrome, aplastic anemia, atopic eczema, atopic dermatitis, autoimmune dermatitis, autoimmune disorder associated with streptococcus infection, autoimmune enteropathy, autoimmune hearing loss, autoimmune lymphoproliferative syndrome (ALPS), autoimmune myocarditis, autoimmune premature ovarian failure, blepharitis, bronchiectasis, bullous pemphigoid, cardiovascular disease, catastrophic antiphospholipid syndrome, celiac disease, cervical spondylosis, chronic ischemia, cicatricial pemphigoid, clinically isolated syndrome (cis) with risk for multiple sclerosis, childhood onset psychiatric disorder, dacryocystitis, dermatomyositis, diabetic retinopathy, disk herniation, disk prolaps, drug induced immune hemolytic anemia, endometriosis, endophthalmitis, episcleritis, erythema multiforme, erythema multiforme major, gestational pemphigoid, Guillain-Barré syndrome (GBS), Hughes syndrome, idiopathic Parkinson's disease, idiopathic interstitial pneumonia, IgE-mediated allergy, immune hemolytic anemia, inclusion body myositis, infectious ocular inflammatory disease, inflammatory demyelinating disease, inflammatory heart disease, inflammatory kidney disease, IPF/UIP, iritis, keratitis, keratojuntivitis sicca, Kussmaul disease or Kussmaul-Meier disease, Landry's paralysis, Langerhan's cell histiocytosis, livedo reticularis, macular degeneration, microscopic polyangiitis, morbus bechterev, motor neuron disorders, mucous membrane pemphigoid, multiple organ failure, myasthenia gravis, myelodysplastic syndrome, myocarditis, nerve root disorders, neuropathy, non-A non-B hepatitis, optic neuritis, osteolysis, pauciarticular JRA, peripheral artery occlusive disease (PAOD), peripheral vascular disease (PVD), peripheral artery, disease (PAD), phlebitis, polyarteritis nodosa (or periarteritis nodosa), polychondritis, poliosis, polyarticular JRA, polyendocrine deficiency syndrome, polymyositis, polymyalgia rheumatica (PMR), primary Parkinsonism, prostatitis, pure red cell aplasia, primary adrenal insufficiency, recurrent neuromyelitis optica, restenosis, rheumatic heart disease, sapho (synovitis, acne, pustulosis, hyperostosis, and osteitis), secondary amyloidosis, shock lung, scleritis, sciatica, secondary adrenal insufficiency, silicone associated connective tissue disease, sneddon-wilkinson dermatosis, spondilitis ankylosans, Stevens-Johnson syndrome (SJS), temporal arteritis, toxoplasmic retinitis, toxic epidermal necrolysis, transverse myelitis, TRAPS (tumor necrosis factor receptor, type I allergic reaction, type II diabetes, urticaria, usual interstitial pneumonia (UIP), vasculitis, vernal conjunctivitis, viral retinitis, Vogt-Koyanagi-Harada syndrome (VKH syndrome), wet macular degeneration, and wound healing. 
     
     
         84 . The method of  claim 83 , wherein the administering is parenteral, subcutaneous, intramuscular, intravenous, intrarticular, intrabronchial, intraabdominal, intracapsular, intracartilaginous, intracavitary, intracelial, intracerebellar, intracerebroventricular, intracolical, intracervical, intragastric, intrahepatic, intramyocardial, intraosteal, intrapelvic, intrapericardial, intraperitoneally, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravesical, by bolus, vaginal, rectal, buccal, sublingual, intranasal, or transdermal. 
     
     
         85 . A method of determining the presence, amount or concentration of at least one target or fragment thereof in a test sample by an immunoassay,
 wherein the method comprises the step of contacting the test sample with at least one binding protein and at least one detectable label,   and wherein the at least one binding protein comprises the binding protein of  claim 61 .   
     
     
         86 . The method of  claim 85 , further comprising the steps of:
 contacting the test sample with the at least one binding protein, wherein the binding protein binds to an epitope on the target or fragment thereof so as to form a first complex;   contacting the complex with the at least one detectable label, wherein the detectable label binds to the binding protein or an epitope on the target or fragment thereof that is not bound by the binding protein to form a second complex; and   detecting the presence, amount, or concentration of the target or fragment thereof in the test sample based on the signal generated by the detectable label in the second complex, wherein the presence, amount, or concentration of the target or fragment thereof is directly correlated with the signal generated by the detectable label.   
     
     
         87 . The method of  claim 85 , further comprising the steps of:
 contacting the test sample with the at least one binding protein, wherein the binding protein binds to an epitope on a target or fragment thereof so as to form a first complex;   contacting the complex with the at least one detectable label, wherein the detectable label competes with the target or fragment thereof for binding to the binding protein so as to form a second complex; and   detecting the presence, amount, or concentration of the target or fragment thereof in the test sample based on the signal generated by the detectable label in the second complex, wherein the presence, amount, or concentration of the target or fragment thereof is indirectly correlated with the signal generated by the detectable label.   
     
     
         88 . A kit for assaying a test sample for the presence, amount, or concentration of the target or fragment thereof, the kit comprising
 (a) instructions for assaying the test sample for the target or fragment thereof, and   (b) at least one binding protein comprising the binding protein of  claim 61 .