Fc fusion
The present invention relates to a simple method for generating antibody-based structures suitable for in vivo use. In particular, the invention relates to a method for the generation of antibody-based structures suitable for in vivo use comprising the steps of: (a) selecting an antibody single variable domain having an epitope binding specificity; and (b) attaching the single domain of step (a) to an effector group. Uses of molecules generated using the method of the Invention are also described.
1 . A method for synthesising a single-domain-effector group (dAb-effector group) suitable for in vivo use comprising the steps of:
(a) selecting an antibody single variable domain having an epitope binding specificity; and
(b) attaching the single domain of step (a) to an effector group.
2 . A method according to claim 1 wherein the antibody single variable domain is a heavy chain variable domain.
3 . A method according to claim 1 wherein the antibody single variable domain is a light chain variable domain.
4 . A method according to claim 3 wherein the light chain variable domain is a member of the Vκ sub-group of domains.
5 . A method according to claim 3 wherein the light chain variable domain is a member of the Vλ sub-group of domains.
6 . A method according to claim 1 wherein the effector group comprises any one or more of those groups selected from the group consisting of: an antibody light chain constant region (C L ), an antibody CH1 heavy chain domain, an antibody CH2 heavy chain domain, an antibody CH3 heavy chain domain, an Fc region of an antibody and a binge region of an antibody molecule.
7 . A method according to claim 1 , wherein the effector group constitutes an Fc region of an antibody.
8 . A method according to claim 1 , wherein the effector group consists of a CH2 and CH3 domain.
9 . A method according to claim 6 , wherein the effector group consists of a CH2 domain, a CH3 domain and the binge region of an antibody molecule.
10 . A method according to claim 1 , wherein the antibody single variable domain is a non-Camelid variable domain.
11 . A method according to claim 10 , wherein the antibody single variable domain is a human variable domain.
12 . A method according to claim 1 , wherein the antibody single variable domain comprises one or more human framework regions.
13 . A method according to claim 1 , wherein the antibody single variable domain comprises four framework regions as defined by Kabat, which are derived from a human.
14 . A method according to claim 13 , wherein one or more of the human framework regions as defined by Kabat are identical on the amino acid level to those encoded by human germline antibody genes.
15 . A method according to claim 1 , wherein the antibody single variable domain is isolated, in part, by human immunisation.
16 . A method according to claim 1 , wherein the antibody single variable domain is not isolated by animal immunisation.
17 . A method according to claim 1 , wherein the antibody single variable domain binds to the superantigens protein A or protein L.
18 . A method according to claim 1 , wherein the effector group is of Camelid or human origin.
19 . A method according to claim 1 , wherein the single variable domain comprises one or more human framework regions and the immunoglobulin effector group is of human origin.
20 . A method according to claim 19 , wherein the single variable domain comprises four human framework regions and the immunoglobulin effector group is of human origin.
21 . A method according to claim 1 , wherein attaching of the single variable domain to the effector group in step (b) is effected by expressing the single-domain-effector group as a fusion polypeptide.
22 . A dAb-effector group comprising:
(a) an antibody single variable domain having an epitope binding specificity; and
(b) an effector group attached to said antibody single variable domain.
23 . A medicament comprising the dAb-effector group of claim 22 .
24 . A dAb-effector group according to claim 22 , wherein the antibody single variable domain is a heavy chain variable domain.
25 . A dAb-effector group according to claim 22 wherein the antibody single variable domain is a light chain variable domain.
26 . A dAb-effector group according to claim 25 wherein the light chain variable domain is a member of the Vκ sub-group of domains.
27 . A dAb-effector group according to claim 25 wherein the light chain variable domain is a member of the Vλ sub-group of domains.
28 . A dAb-effector group according to claim 22 , wherein the effector group comprises any one or more of those groups selected from the group consisting of: an antibody light chain constant region (C L ), an antibody CH1 heavy chain domain, an antibody CH2 heavy chain domain, an antibody CH3 heavy chain domain, an Fe region of an antibody and a hinge region of an antibody molecule.
29 . A dAb-effector group according to claim 28 wherein the effector group consists of a CH2 and CH3 domain.
30 . A dAb-effector group according to claim 28 wherein the effector group consists of a CH2 domain, a CH3 domain and the hinge region of an antibody molecule.
31 . A dAb-effector group according to claim 28 wherein the effector group constitutes an Fc region of an antibody.
32 . A dAb-effector group according to claim 22 , wherein the antibody single variable domain is of human origin.
33 . A dAb-effector group according to claim 22 , wherein the antibody single variable domain comprises human framework regions.
34 . A dAb-effector group according to claim 22 , wherein the effector group is of Camelid or human origin.
35 . A dAb-effector group according to claim 22 , wherein the single variable domain comprises one or more human framework regions and the immunoglobulin effector group is of human origin.
36 . Two or more dAb-effector groups according to claim 22 provided as a higher order structure selected from the group consisting of the following: dimers, trimers and multimers.
37 . Two dAb-effector groups according to claim 36 provided as a heterodimer or a homodimer.
38 . Two dAb-effector groups according to claim 37 provided as a homodimer.
39 . A nucleic acid molecule encoding a dAb-effector group according to claim 22 .
40 . A nucleic acid molecule according to claim 39 further encoding a signal sequence for export of the dAb and effector group from the cytoplasm of a host cell upon expression.
41 . A vector comprising nucleic acid according to claim 39 .
42 . A host cell transfected with a vector according to claim 41 .
43 . A composition comprising a dAb-effector group(s) according to claim 22 and a pharmaceutically acceptable carrier, diluent or excipient.
44 . A composition according to claim 43 having a t1/2 alpha of 15 minutes or more.
45 . A composition according to claim 43 having a t1/2 alpha from 1 to 6 hours.
46 . A composition according to claim 43 having a t1/2 beta of 2.5 hours or more.
47 . A composition according to claim 43 having a t1/2 beta of 1 day or more
48 . A composition according to claim 47 having a t1/2 beta of 2 days or more.
49 . A composition according to claim 48 having a t1/2 beta of 3 days or more.
50 . A composition according to claim 43 having an AUC of 1 mg.min/ml or more.
51 . A composition according to claim 50 having an AUC from 15 to 150 mg.min/ml.
52 . A method of treating and/or preventing disease in a patient, wherein the method comprises administering to the patient a dAb-effector group(s) according to claim 22 or a composition according to claims 43 .
53 . A medicament for the treatment and/or prevention of disease, comprising the dAb-effector group of claim 22 or the composition of claim 43 .
54 . A method for the treatment and/or prophylaxis of an inflammatory disease in a patient in need of such treatment and/or prophylaxis which comprises the step of administering to that patient a therapeutically effective amount of a dAb-effector group according to claim 22 .
55 . A method according to claim 54 wherein the inflammatory disease is mediated by TNF alpha and is selected from the group consisting of the following: rheumatoid arthritis, psoriasis, Crohns disease, inflammatory bowel disease (IBD), multiple sclerosis, septic shock, Alzheimer's, coronary thrombosis, chronic obstructive pulmonary disease (COPD) and glomerular nephritis.
56 . A method for reducing and/or preventing and/or suppressing cachexia in a patient which is mediated by TNF alpha which method comprises the step of administering to a patient in need of such treatment a therapeutically effective amount of a dAb-effector group according to claim 22 .
57 . A method according to claim 56 , wherein the TNF alpha is human TNF alpha and the patient is a human.
58 . A method according to claim 54 to 56 wherein the dAb-effector group is TAR1-5-19-effector group.
59 . A method or a use according to claim 58 wherein the effector group is Fc.
60 . A method or a use according to claim 54 to 56 wherein the dAb-effector group is administered in a dosage range of 0.5 to 20 mg/Kg.
61 . A method or a use according to claim 60 wherein the dAb-effector group is administered in a dose of range of 1 to 10 mg/Kg.