Binding members to tnf alpha

Abstract

The present invention relates to anti-TNF alpha binding members and in particular to monovalent, high potency TNF alpha-binding antibody fragments being highly stable and soluble. Such binding members may be used in the treatment of inflammatory and other diseases as well as in diagnostics. Also provided are related nucleic acids, vectors, cells, and compositions.

1 - 20 . (canceled) 
     
     
         21 . A method of treating a TNF alpha-mediated disease, the method comprising administering to a subject in need thereof an antibody or fragment thereof having a binding specificity to TNF alpha, the antibody or fragment thereof comprising
 (i) the variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID Nos: 6, 7 and 8; and   (ii) the variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID Nos: 3, 4 and 5.   
     
     
         22 . The method of  claim 21 , wherein the TNF alpha-mediated disease is at least one of proliferative diabetic retinopathy, gouty arthritis, acute gouty arthritis and chronic gouty arthritis, Schnitzler syndrome, systemic juvenile idiopathic arthritis, rheumatoid arthritis, urticaria, vasculitis, type 1 diabetes, type 2 diabetes, recurrent multifocal osteomyelitis, relapsing polychondritis, cyropyrin-associated periodic syndrome (CAPS), Behçet's disease, familial mediterranean fever, chronic obstructive pulmonary disease, polymyalgia rheumatic, NALP3-mutations, pyoderma gangrenosum, chronic idiopathic urticaria, osteoarthritis, wet age-related macular degeneration, dry eye syndrome, psoriasis, synovitis-acne-pustulosis-hyperostosis-osteitis syndrome, macrophage activation syndrome, periodic fever, adenitis, pharyngitis, aphthous ulcer syndrome, adult-onset Still's disease, mevalonate kinase deficiency, uveitis, inflammatory bowel disease, atherosclerosis, TNF-receptor associated periodic syndrome (TRAPS), ankylosing spondylitis, hidradenitis suppurativa, psoriasis, and acne vulgaris. 
     
     
         23 - 35 . (canceled) 
     
     
         36 . The method of  claim 21 , wherein the antibody or fragment thereof has an IC 50  with regard to human TNF alpha of lower than 50 pM. 
     
     
         37 . The method of  claim 21 , wherein the antibody or fragment thereof is humanized. 
     
     
         38 . The method of  claim 21 , wherein the antibody or fragment thereof comprises
 (i) a variable light chain as set forth in SEQ ID No. 1; and/or   (ii) a variable heavy chain as set forth in SEQ ID No. 2.   
     
     
         39 . The method of  claim 21 , wherein the antibody or fragment thereof further comprises a linker sequence. 
     
     
         40 . The isolated nucleic acid molecule of  claim 39 , wherein the linker sequence is the sequence set forth in SEQ ID No: 10. 
     
     
         41 . The method of  claim 21 , wherein the antibody or fragment thereof comprises SEQ ID No. 9. 
     
     
         42 . The method of  claim 21 , wherein the variable heavy chain of the antibody or fragment thereof further comprises at least one of the following residues:
 (i) Serine (S) at heavy chain amino acid position 12 (according to AHo numbering);   (ii) Serine (S) or Threonine (T) at heavy chain amino acid position 103 (according to AHo numbering); and/or   (iii) Serine (S) or Threonine (T) at heavy chain amino acid position 144 (according to AHo numbering).   
     
     
         43 . The method of  claim 21 , wherein the antibody or fragment thereof is
 (i) a Fab, a Fab′, a F(ab)′ 2 , a scFv, or a Fv fragment, or;   (ii) a full-length immunoglobulin molecule.   
     
     
         44 . The method of  claim 21 , wherein the antibody or fragment thereof is monovalent or multivalent, wherein the binding member is optionally bispecific. 
     
     
         45 . The method of  claim 21 , wherein the antibody or fragment thereof is a monovalent antibody or fragment thereof. 
     
     
         46 . The method of  claim 45 , wherein the antibody or fragment thereof is a scFv. 
     
     
         47 . The method of  claim 21 , wherein the antibody or fragment thereof is a diabody, a single-chain diabody or a tandem scFv. 
     
     
         48 . The method of  claim 21 , wherein the antibody or fragment thereof remains at least 93% monomeric after incubation for 1 week at 37° C. at a concentration of 10 mg/ml PBS pH7.2.