MODIFIED PROTEINS AND PEPTIDES
The present invention relates to modified proteins and peptides that have reduced ability to bind to pre-existing antibodies. Such modified protein/peptide molecules can comprise C-terminal additions, extensions or tags and/or certain amino acid substitutions. Such modified molecules (e.g. fusions and conjugates) comprise proteins, peptides, antigen binding molecules, antibodies or antibody fragments such as single variable domains e.g. human immunoglobulin (antibody) single variable domains, and also single variable domains derived from non-human sources such as a llama or camel, e.g. a VHH including a Nanobody™ (described in e.g. WO 94/04678 and WO 95/04079 inter alia). The invention further relates to uses, formulations, compositions comprising such modified C terminally extended and/or amino acid substituted molecules and also to methods of production and expression of these molecules.
1 . A single immunoglobulin variable domain (dAb), wherein said dAb comprises a sequence selected from a sequence having at least 90% sequence identity to SEQ ID NO 4; a sequence having at least 90% sequence identity to SEQ ID NO 5; and a sequence having at least 90% sequence identity to SEQ ID NO 6; and wherein said dAb comprises
a C-terminal extension which comprises an amino acid extension of from one amino acid to 5 amino acids.
2 . The single immunoglobulin variable domain (dAb) according to claim 1 , wherein said dAb is derived from a Camelid heavy chain (VHH).
3 . The single immunoglobulin variable domain (dAb) according to claim 1 , which comprises a C-terminal extension of from one amino acid to 4 amino acids.
4 . The single immunoglobulin variable domain (dAb) according to claim 1 , wherein said C-terminal extension comprises an alanine residue.
5 . The single immunoglobulin variable domain (dAb) according to claim 1 , wherein said C terminal extension comprises an amino acid extension selected from: (a) A (b) AS, (c) AST (d) ASTK, (e) ASTKG (f) AAA or (g) T.
6 - 24 . (canceled)
25 . A pharmaceutical composition comprising the dAb according to claim 1 in combination with a pharmaceutically or physiologically acceptable carrier, excipient or diluent.
26 - 39 . (canceled)
40 . A method of treating or preventing an inflammatory disease or disorder, said method comprising administering to a subject a therapeutically or prophylactically effective amount of the dAb according to claim 1 .
41 . The method according to claim 40 , wherein the dAb has an extension of a single alanine at the C terminus.
42 . The method of claim 40 , wherein said dAb comprises a sequence set forth in SEQ ID NO: 4 or SEQ ID NO:5, and wherein said inflammatory disease or disorder is arthritis.
43 . The method of claim 40 , wherein said dAb comprises a sequence set forth in SEQ ID NO:6, and wherein said inflammatory disease or condition is Von Willebrand's disease.
44 . The method according to claim 40 , wherein said administering comprises subcutaneous, intravenous or intramuscular injection.
45 . The method according to claim 40 , wherein said administering comprises parenteral, oral, rectal, transmucosal, ocular, pulmonary or GI tract delivery.
46 - 59 . (canceled)
60 . The single immunoglobulin variable domain (dAb) of claim 1 , wherein said dAb comprises:
a) a sequence set forth in SEQ ID NO 4, and
b) a single alanine C terminal extension.
61 . The single immunoglobulin variable domain (dAb) of claim 1 , wherein said dAb comprises:
a) a sequence set forth in SEQ ID NO 5, and
b) a single alanine C terminal extension.
62 . The single immunoglobulin variable domain (dAb) of claim 1 , wherein said dAb comprises:
a) a sequence set forth in SEQ ID NO 6, and
b) a single alanine C terminal extension.
63 . A single immunoglobulin variable domain (dAb) comprising a sequence set forth in SEQ ID NO 7.
64 . The single immunoglobulin variable domain (dAb) of claim 63 , wherein said dAb consists of the sequence set forth in SEQ ID NO 7.
65 . A single immunoglobulin variable domain (dAb) comprising a sequence set forth in SEQ ID NO 8.
66 . The single immunoglobulin variable domain (dAb) of claim 65 , wherein said dAb consists of the sequence set forth in SEQ ID NO 8.
67 . A single immunoglobulin variable domain (dAb) comprising a sequence set forth in SEQ ID NO 9.
68 . The single immunoglobulin variable domain (dAb) of claim 67 , wherein said dAb consists of the sequence set forth in SEQ ID NO 9.