U.S. patent application number 12/861205 was filed with the patent office on 2011-03-03 for nogo-a neutralizing immunoglobulins for the treatment of neurological diseases.
Invention is credited to JONATHAN HENRY ELLIS, PAUL ANDREW HAMBLIN, ALAN PETER LEWIS, PAUL ALEXANDER WILSON.
Application Number | 20110054149 12/861205 |
Document ID | / |
Family ID | 34712703 |
Filed Date | 2011-03-03 |
United States Patent
Application |
20110054149 |
Kind Code |
A1 |
ELLIS; JONATHAN HENRY ; et
al. |
March 3, 2011 |
NOGO-A Neutralizing Immunoglobulins for the Treatment of
Neurological Diseases
Abstract
The present invention relates to antibodies to NOGO,
pharmaceutical formulations containing them and to the use of such
antibodies in the treatment and/or prophylaxis of neurological
diseases/disorders.
Inventors: |
ELLIS; JONATHAN HENRY;
(STEVENAGE HERTFORDSHIRE, GB) ; HAMBLIN; PAUL ANDREW;
(STEVENAGE HERTFORDSHIRE, GB) ; LEWIS; ALAN PETER;
(STEVENAGE HERTFORDSHIRE, GB) ; WILSON; PAUL
ALEXANDER; (STEVENAGE HERTFORDSHIRE, GB) |
Family ID: |
34712703 |
Appl. No.: |
12/861205 |
Filed: |
August 23, 2010 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11177648 |
Jul 6, 2005 |
7780964 |
|
|
12861205 |
|
|
|
|
PCT/GB04/05325 |
Dec 20, 2004 |
|
|
|
11177648 |
|
|
|
|
Current U.S.
Class: |
530/388.2 |
Current CPC
Class: |
A61P 9/00 20180101; A61P
25/00 20180101; C12N 5/0618 20130101; A61P 25/28 20180101; C07K
16/18 20130101; C07K 2317/56 20130101; A61P 25/16 20180101; C07K
16/22 20130101; C12N 2501/998 20130101; C07K 2317/76 20130101; C12N
2501/70 20130101; A61K 2039/505 20130101; A61P 25/14 20180101; C07K
2317/24 20130101; A61P 25/02 20180101 |
Class at
Publication: |
530/388.2 |
International
Class: |
C07K 16/18 20060101
C07K016/18 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 22, 2003 |
GB |
0329684.5 |
Dec 22, 2003 |
GB |
0329711.6 |
Claims
1.-30. (canceled)
31. A monoclonal antibody which binds to and neutralises human NOGO
A, the monoclonal antibody comprising a heavy chain variable region
which has the sequence of SEQ ID NO:77, with the exception that the
amino acid residue at position 48 of SEQ ID NO:77 is substituted
with isoleucine, the amino acid residue at position 67 of SEQ ID
NO:77 is substituted with lysine and the amino acid residues at
position 68 and 79 of SEQ ID NO:77 are substituted with alanine; or
an analog of said antibody, wherein the analog comprises an amino
acid modification within a CDR of said heavy chain.
32. The monoclonal antibody of claim 31, wherein said analog
comprises an amino acid modification within CDRH1 of said heavy
chain.
33. The monoclonal antibody of claim 31, wherein said analog
comprises an amino acid modification within CDRH2 of said heavy
chain.
34. The monoclonal antibody of claim 31, wherein said analog
comprises an amino acid modification within CDRH3 of said heavy
chain.
35. The monoclonal antibody of claim 31, comprising a light chain
variable region which has the sequence of SEQ ID NO:23.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 11/177,648, filed Jul. 6, 2005, now U.S. Pat. No. 7,780,964,
which is a continuation in part of International application No.
PCT/GB04/05325, filed Dec. 20, 2004, which claims priority from
United Kingdom application Nos. 0329711.6 and 0329684.5 both filed
on Dec. 22, 2003. The contents of all of the above-referenced
applications are herein incorporated by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to immunoglobulins,
particularly antibodies that bind to NOGO and neutralise the
activity thereof, polynucleotides encoding such antibodies,
pharmaceutical formulations containing said antibodies and to the
use of such antibodies in the treatment and/or prophylaxis of
neurological diseases.
BACKGROUND OF THE INVENTION
[0003] Stroke is a major cause of death and disability in the
Western World. There is no approved therapy for the treatment of
stroke other than tissue plasminogen (t-PA) which has to be
administered within 3 hours of onset following a computer
tomography (CT) scan to exclude haemorrhage. To date most
therapeutic agents directed towards the treatment of acute stroke
(i.e. neuroprotection), have predominantly involved targeting
glutamate receptors and their down stream signalling pathways known
to be involved in acute cell death. However to date these
strategies have proved unsuccessful in clinical trials and are
often associated with dose-limiting side effects (Hill &
Hachinski, The Lancet, 352: (suppl III) 10-14 (1998)). Therefore
there is a need for novel approaches directed towards the
amelioration of cell death following the cessation of blood flow.
Neuroprotection is the ability of a treatment to prevent or
ameliorate neuronal cell loss in response to an insult or disease
process. This maybe achieved by targeting the neurons directly or
indirectly by preventing glial (including oligodendrocyte) cell
loss.
[0004] Following the onset of stroke, some degree of spontaneous
functional recovery is observed in many patients, suggesting that
the brain has the (albeit limited) ability to repair and/or remodel
following injury. Agents that have the potential to enhance this
recovery may therefore allow intervention to be made much later
(potentially days) following the onset of cerebral ischaemia.
Agents which are able to offer both acute neuroprotection and
enhance functional recovery may provide significant advantages over
current potential neuroprotective strategies.
[0005] Alzheimer's disease (AD) is characterised by the presence of
two diagnostic features of pathology. These are amyloid plaques and
neurofibrillary tangles composed of aggregated beta-amyloid peptide
(A.beta.40 and A.beta.42) and hyperphosphorylated tau respectively
(Dawbarn & Allen 2001 Neurobiology of Alzheimer's Disease
OUP).
[0006] A comprehensive study has shown a strong link in patients
between beta-amyloid accumulation and cognitive decline (Naslund et
al, JAMA, March 22/29, 2000, Vol. 283, No; 12, page 1571-1577).
This is consistent with genetic and epidemiological studies that
suggest that some mutations in APP and presenilin genes can
predispose to early onset AD, which mutations also enhance the
levels of A.beta.40 and A.beta.42 peptide, including the ratio
thereof.
[0007] Cleavage of the type I transmembrane amyloid precursor
protein (APP) by two distinct proteases designated beta- and
gamma-secretase is necessary for the formation of beta-amyloid
peptide. The molecular identity of beta-secretase as the
aspartyl-protease Asp2/BACE1 has been confirmed (Hussain et al Mol.
Cell. NeuroSci. 16, 609-619 (2000); Vassar et al, Science (1999),
Oct. 22; 286 (5440):735-741). The nature of gamma-secretase remains
the source of some debate and is likely to consist of a high
molecular weight complex consisting of at least the following
proteins: presenilins, Aph1, Pen2 and nicastrin (reviewed in Medina
& Dotti Cell Signalling 2003 15(9):829-41).
[0008] The processing of APP within the CNS is likely to occur
within a number of cell-types including neurons, oligodendrocytes,
astrocytes and microglia. While the overall rate of APP processing
in these cells will be influenced by the relative level of
expression of APP, BACE1/Asp2, presenilin-1 and -2, Aph1, Pen2 and
nicastrin.
[0009] Furthermore, additional factors regulating the subcellular
location of APP can also influence its processing as shown by the
finding that mutation of the YENP motif in the APP cytoplasmic
domain which blocks its endocytosis reduces beta-amyloid production
(Perez et al 1999 J Biol Chem 274 (27) 18851-6). Retention of the
APP-beta-CTF in the ER by the addition of the KKQN retention motif
is sufficient to reduce amyloid production in transfected cells
(Maltese et al 2001 J Biol Chem 276 (23) 20267-20279). Conversely,
elevation of endocytosis, by overexpression of RabS is sufficient
to elevate amyloid secretion from transfected cells (Grbovic et al
2003 J Biol Chem 278 (33) 31261-31268).
[0010] Consistent with these findings further studies have shown
that reduction of cellular cholesterol levels (a well known risk
factor for AD) reduced beta-amyloid formation. This change was
dependent on altered endocytosis as demonstrated by the use of the
dominant negative dynamin mutants (K44A) and overexpression of the
Rab5 GTPase activating protein RN-Tre (Ehehalt et al 2003 J Cell
Biol 160 (1) 113-123).
[0011] Cholesterol rich microdomains or rafts are also an important
cellular site of beta-amyloid production and APP, BACE1 and
components of the gamma-secretase complex have all been shown to
transiently reside within rafts. Antibody cross-linking of APP and
BACE1 towards cholesterol rich rafts was able to elevate
beta-amyloid production (Ehehalt et al 2003 J Cell Biol 160 (1)
113-123). Expression of GPI-anchored BACE1, which is exclusively
targeted to lipid rafts, is similarly able to elevate APP cleavage
and beta-amyloid production (Cordy et al 2003 PNAS 100(20)
11735-11740).
[0012] The mechanisms underlying functional recovery are currently
unknown. The sprouting of injured or non-injured axons has been
proposed as one possible mechanism. However, although in vivo
studies have shown that treatment of spinal cord injury or stroke
with neurotrophic factors results in enhanced functional recovery
and a degree of axonal sprouting, these do not prove a direct link
between the degree of axonal sprouting and extent of functional
recovery (Jakeman, et al. 1998, Exp. Neurol. 154: 170-184, Kawamata
et al. 1997, Proc Natl Acad. Sci. USA., 94:8179-8184, Ribotta, et
al. 2000, J Neurosci. 20: 5144-5152). Furthermore, axonal sprouting
requires a viable neuron. In diseases such as stroke which is
associated with extensive cell death, enhancement of functional
recovery offered by a given agent post stroke may therefore be
through mechanisms other than axonal sprouting such as
differentiation of endogenous stem cells, activation of redundant
pathways, changes in receptor distribution or excitability of
neurons or glia (Fawcett & Asher, 1999, Brain Res. Bulletin,
49: 377-391, Horner & Gage, 2000, Nature 407 963-970).
[0013] The limited ability of the central nervous system (CNS) to
repair following injury is thought in part to be due to molecules
within the CNS environment that have an inhibitory effect on axonal
sprouting (neurite outgrowth). CNS myelin is thought to contain
inhibitory molecules (Schwab M E and Caroni P (1988) J. Neurosci.
8, 2381-2193). Two myelin proteins, myelin-associated glycoprotein
(MAG) and NOGO have been cloned and identified as putative
inhibitors of neurite outgrowth (Sato S. et al (1989) Biochem.
Biophys. Res. Comm.163, 1473-1480; McKerracher L et al (1994)
Neuron 13, 805-811; Mukhopadhyay G et al (1994) Neuron 13, 757-767;
Torigoe K and Lundborg G (1997) Exp. Neurology 150, 254-262;
Schafer et al (1996) Neuron 16, 1107-1113; WO9522344; WO9701352;
Prinjha R et al (2000) Nature 403, 383-384; Chen M S et al (2000)
Nature 403, 434-439; GrandPre T et al (2000) Nature 403, 439-444;
US005250414A; WO200005364A1; WO0031235).
[0014] Three forms of human NOGO have been identified: NOGO-A
having 1192 amino acid residues (GenBank accession no. AJ251383);
NOGO-B, a splice variant which lacks residues 186 to 1004 in the
putative extracellular domain (GenBank accession no. AJ251384) and
a shorter splice variant, NOGO-C, which also lacks residues 186 to
1004 and also has smaller, alternative amino terminal domain
(GenBank accession no. AJ251385) (Prinjha et al (2000) supra).
[0015] Inhibition of the CNS inhibitory proteins such as NOGO may
provide a therapeutic means to ameliorate neuronal damage and
promote neuronal repair and growth thereby potentially assisting
recovery from neuronal injury such as that sustained in stroke.
Examples of such NOGO inhibitors may include small molecules,
peptides and antibodies.
[0016] Antibodies typically comprise two heavy chains linked
together by disulphide bonds and two light chains. Each light chain
is linked to a respective heavy chain by disulphide bonds. Each
heavy chain has at one end a variable domain followed by a number
of constant domains. Each light chain has a variable domain at one
end and a constant domain at its other end. The light chain
variable domain is aligned with the variable domain of the heavy
chain. The light chain constant domain is aligned with the first
constant domain of the heavy chain. The constant domains in the
light and heavy chains are not involved directly in binding the
antibody to antigen.
[0017] The variable domains of each pair of light and heavy chains
form the antigen binding site. The variable domains on the light
and heavy chains have the same general structure and each domain
comprises a framework of four regions, whose sequences are
relatively conserved, connected by three complementarity
determining regions (CDRs) often referred to as hypervariable
regions. The four framework regions largely adopt a beta-sheet
conformation and the CDRs form loops connecting, and in some cases
forming part of, the beta-sheet structure. The CDRs are held in
close proximity by the framework regions and, with the CDRs from
the other domain, contribute to the formation of the antigen
binding site. CDRs and framework regions of antibodies may be
determined by reference to Kabat et al ("Sequences of proteins of
immunological interest" US Dept. of Health and Human Services, US
Government Printing Office, 1987).
[0018] It has been reported that a murine monoclonal antibody,
IN-1, that was raised against NI-220/250, a myelin protein which is
a potent inhibitor of neurite growth (and subsequently shown to be
fragment of NOGO-A), promotes axonal regeneration (Caroni, P and
Schwab, M E (1988) Neuron 1 85-96; Schnell, L and Schwab, M E
(1990) Nature 343 269-272; Bregman, B S et al (1995) Nature 378
498-501 and Thallmair, M et al (1998) Nature Neuroscience 1
124-131). It has also been reported that NOGO-A is the antigen for
IN-1 (Chen et al (2000) Nature 403 434-439). Administration of IN-1
Fab fragment or humanised IN-1 to rats that have undergone spinal
cord transection, enhanced recovery (Fiedler, M et al (2002)
Protein Eng 15 931-941; Brosamle, C et al (2000) J. Neuroscience 20
8061-8068). However to date there is no evidence in the literature
to suggest that IN-1, or its humanised form, can bind and inhibit
human NOGO-A, a necessary requirement for a monoclonal antibody to
be useful in the therapeutic treatment of NOGO-mediated diseases
and disorders such as stroke and neurodegenerative diseases in
humans.
[0019] Monoclonal antibodies which bind to NOGO are described in WO
04/052932 and WO2005028508. WO 04/052932 discloses a murine
antibody 11C7 which binds to certain forms of human NOGO with high
affinity.
[0020] It is desirable to isolate and develop further
therapeutically useful monoclonal antibodies that bind to, and
inhibit the activity of, human NOGO. The process of
neurodegeneration underlies many neurological diseases/disorders
including, but not limited to, acute diseases such as stroke
(ischemic or haemorrhagic), traumatic brain injury and spinal cord
injury as well as chronic diseases including Alzheimer's disease,
fronto-temporal dementias (tauopathies), peripheral neuropathy,
Parkinson's disease, Creutzfeldt-Jakob disease (CJD),
Schizophrenia, amyotrophic lateral sclerosis (ALS), multiple
sclerosis, Huntington's disease, multiple sclerosis and inclusion
body myositis. Consequently an anti-NOGO monoclonal antibody may be
useful in the treatment of these diseases/disorders. Such
antibodies for the treatment of the above mentioned
disease/disorders are provided by the present invention and
described in detail below.
[0021] The unpublished patent application, with the International
Patent Application number PCT/GB2004/005325, also discloses high
affinity monoclonal antibodies, including a murine monoclonal
antibody 2A10, and a humanised variant thereof H1L11.
SUMMARY OF THE INVENTION
[0022] The present invention provides a number of monoclonal
antibodies that bind to human NOGO. The present application gives
reference to many SEQ ID numbers, which are summarised in Table 12,
with the actual sequences following that table towards the end of
this document.
[0023] The murine antibody 2A10 binds to human NOGO with high
affinity, and binds to the form of NOGO which is expressed by human
cell lines with high affinity. 2A10 has been humanised successfully
in the past (H1L11 which comprises the heavy chain variable region
H1 (SEQ ID NO. 77) and the light chain variable region L11 (SEQ ID
NO. 78)). The present invention provides additional humanised
monoclonal antibodies which retain a high proportion of the
affinity of the donor antibody (2A10) to human NOGO. In particular
the antibodies of the present invention have a high affinity to
human NOGO both in recombinant form as expressed in bacterial cells
(such as E. Coli), and also in the form that it is expressed by a
human neuroblastoma cell line (for example the human neuroblastoma
cell line IMR32).
[0024] For the purposes of providing humanised variants of 2A10,
the present inventors have identified a number of key amino acid
residues in the framework sequence of the 2A10 variable regions
which are believed to be important in optimal retention of binding
affinity to human NOGO. The 2A10 heavy chain variable region (VH)
is provided in SEQ ID NO. 7; the 2A10 light chain variable region
(VL) is provided in SEQ ID NO. 8. Chimeric heavy and light chains
comprising the murine variable regions and human constant regions
are provided in SEQ ID NOs 9 and 10 respectively (the combination
of the two chimeric chains is termed HcLc). The skilled reader will
understand that SEQ ID NOs 9 and 10 represent the heavy chain or
light chains prior to any processing (e.g. host cell mediated
processing) for removal of a signal sequence. Typically the
processed forms of the antibody chains will begin at position 20
(after the removal of the signal sequence (residues 1-19) which
corresponds to SEQ ID NO. 75).
TABLE-US-00001 TABLE 1 CDRs of the 2A10 heavy chain are: CDR
According to Kabat H1 SYWMH (SEQ ID NO: 1) H2 NINPSNGGTNYNEKFKS
(SEQ ID NO: 2) H3 GQGY (SEQ ID NO: 3)
TABLE-US-00002 TABLE 2 CDRs of 2A10 light chain: CDR According to
Kabat L1 RSSKSLLYKDGKTYLN (SEQ ID NO: 4) L2 LMSTRAS (SEQ ID NO: 5)
L3 QQLVEYPLT (SEQ ID NO: 6)
[0025] The CDRs were identified according to Kabat (Kabat et al.
(1991) "Sequences of proteins of immunological interest"; Fifth
Edition; US Department of Health and Human Services; NIH
publication No 91-3242). CDRs preferably are as defined by Kabat
but following the principles of protein structure and folding as
defined by Chothia and Lesk, (Chothia et al., (1989) "Conformations
of immunoglobulin hypervariable regions"; Nature 342, p 877-883) it
will be appreciated that additional residues may also be considered
to be part of the antigen binding region and are thus encompassed
by the present invention.
[0026] The VH and VL domains H1 and L11 have been previously
described in PCT/GB2004/005325, and are a result of the CDRs
mentioned in tables 1 and 2 being grafted into human variable
regions with high homology to the 2A10 donor antibody, each grafted
construct further comprising back mutations in kabat positions 93
and 94 (for the H1 VH) or 4, 45 and 46 (for L11 VL).
[0027] The 2A10 antibody is capable of binding to human NOGO, and
also binds to Marmoset and Rat NOGO, and it is believed that the
new humanised antibodies of the present invention will retain that
property. The sequence of Marmoset NOGO-A fragment is given in SEQ
ID NO. 113.
[0028] Heavy Chain Variable Region (VH)
[0029] In one aspect of the present invention the antibodies
comprise a heavy chain variable region having the amino acid
sequence of SEQ ID NO. 77 (H1 variable region) further comprising a
number of substitutions at one or more of positions 12, 20, 38, 40,
48, 67, 68, 70, 72, 74, 76, 79 and 91; wherein each substituted
amino acid residue is replaced with the amino acid residue at the
equivalent position in SEQ ID NO 7 (the heavy chain variable region
of the donor antibody 2A10) and the number of substitutions is
between 1 and 13. In other embodiments the number of substitutions
is 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11,
or 12, or 13. In other embodiments the number of substitutions is
between 2 and 13, or 3 and 13, or 4 and 13, or 5 and 13, or 6 and
13, or 7 and 13, or 8 and 13, or 9 and 13, or 10 and 13 or 11 and
13, or 12 and 13.
[0030] In this context the substitutions that are described are
equivalent in concept to "back-mutations" where the human framework
amino acid residues in specific positions within the H1 sequence
are back-mutated to the amino acid residues in the equivalent
position within the 2A10 donor antibody sequence.
[0031] Unless specifically stated otherwise, when a numerical
position of an amino acid residue found within a specific sequence
is mentioned in this document, for example "position 12", it is
intended that the skilled reader assigns the first amino acid in
the sequence the position "1" and counts from position one and
identifies the amino acid which is in the desired position, in this
example the twelth amino acid residue in the sequence. The skilled
reader will notice that this numbering system does not correspond
with the Kabat numbering system which is often used for amino acid
positions within antibody sequences. The following table (Table 3)
illustrates the substitutions/back-mutations of the present
invention and gives their numerical positions and the Kabat number
for that numerical position:
TABLE-US-00003 TABLE 3 Corresponding Human framework Kabat number
of residue residue of H1 (SEQ Numerical that numerical in murine
2A10 ID NO 77) Position position (SEQ ID NO 7) K 12 12 V V 20 20 L
R 38 38 K A 40 40 R M 48 48 I R 67 66 K V 68 67 A M 70 69 L R 72 71
V T 74 73 K T 76 75 S V 79 78 A T 91 87 S
[0032] With reference to Table 3, in one embodiment the monoclonal
antibodies of the present invention comprise the
substitution/backmutation at position 79 to form antibodies of
"Class A".
[0033] For optimal binding affinity, it has been found that the
pair of amino acid residues in positions 48 and 68, should be I and
A respectively (as they exist in 2A10) or M and V respectively (as
they exist in H1). Accordingly, again with reference to Table 3, in
another embodiment, "Class B", the monoclonal antibody comprises
the substitution at positions 79, 48 and 68.
[0034] In another embodiment ("Class C") the monoclonal antibodies
of "Class A" or "Class B" further comprise a substitution at
positions 40 and/or 67.
[0035] In another embodiment the "Class C" monoclonal antibodies of
the present invention further comprise a substitution at positions
38, 72 or 70 to form "Class D" antibodies.
[0036] In another embodiment the "Class D" monoclonal antibodies
further comprise substitutions at one or more of positions 12, 20,
74, 76 or 91 ("Class E").
[0037] The following table includes details of 20 different heavy
chain variable (VH) regions which may form part of the antibodies
of the present invention. Each of the disclosed VH is based on the
H1 VH (SEQ ID NO. 77) further comprising the substitutions
mentioned in the table (Table 4) where the H1 residue at the
relevant position is substituted with the 2A10 residue at that
position (in the table, "-" means that there is no substitution in
that position, and so the residue remains as in the sequence of
H1):
TABLE-US-00004 TABLE 4 Numerical Residue No. 12 20 38 40 48 67 68
70 72 74 76 79 91 Kabat No. 12 20 38 40 48 66 67 69 71 73 75 78 87
2A10 New VH V L K R I K A L V K S A S (SEQ ID H1 NO. X) K V R A M R
V M R T T V T H5 (11) -- -- -- -- -- -- -- -- -- -- -- A -- H6 (12)
-- -- -- -- I -- A -- -- -- -- A -- H700 (13) -- -- -- -- I -- A L
V K -- A -- H8 -- -- -- -- I -- A L V -- -- A -- H9 -- L -- -- I --
A L V K -- A -- H10 -- L K -- I -- A L V K -- A -- H12 -- -- -- --
I -- A L -- K -- -- -- H13 -- -- -- -- I -- A -- V -- -- A -- H14
(14) V L K -- I -- A L -- -- -- A -- H15 (15) V L K R I K A L V --
S A S H16 (16) -- -- K R I K A L V K -- A -- H17 (17) V L K R I K A
L V K -- A -- H18 (18) V L K R I K A L V K S A S H19 (85) -- -- --
R I -- A -- -- -- -- A -- H20 (86) -- -- -- -- I K A -- -- -- -- A
-- H21 (87) -- -- -- R I K A -- -- -- -- A -- H22 (88) -- -- K R I
K A -- -- -- -- A -- H23 (89) -- -- K R I K A -- V -- -- A -- H24
(90) -- -- K R I K A L V -- -- A -- H25 (91) -- -- -- R -- -- -- --
-- -- -- A --
[0038] Accordingly there is provided a monoclonal antibody
comprising the heavy chain variable region having a sequence given
in any one of SEQ ID NOs: 11-18 or 85-91. In another embodiment
there is provided a monoclonal antibody comprising a heavy chain
having a sequence given in any one of SEQ ID NOs: 26-33 or
92-98.
[0039] In a particular embodiment the antibody comprises the VH
regions provided in SEQ ID NOs 11, 12, 16, 18, 85, 86, 87 or 91 or
the heavy chains provided in SEQ ID NOs 26, 27, 31, 33, 92, 93, 94
or 98.
[0040] Light Chain Variable Region
[0041] In one aspect of the present invention the antibodies
comprise a light chain variable region having the amino acid
sequence of SEQ ID NO. 20 (L13 variable region) optionally further
comprising a number of substitutions at one or more of positions 4,
7, 11, 19, 42, 64 and 70; wherein each substituted amino acid
residue is replaced with the amino acid residue at the equivalent
position in SEQ ID NO. 8 (the light chain variable region of the
donor antibody 2A10) and the number of substitutions is between 0
and 7. In other embodiments the number of substitutions is 1, or 2,
or 3, or 4, or 5, or 6, or 7. In other embodiments the number of
substitutions is between 2 and 7, or 3 and 7, or 4 and 7, or 5 and
7, or 6 and 7.
[0042] The following table (Table 5) illustrates the
substitutions/back-mutations of the present invention and gives
their numerical positions and the Kabat number for that numerical
position:
TABLE-US-00005 TABLE 5 Human framework Corresponding residue of L13
Numerical Kabat residue in murine 2A10 (SEQ ID NO 20) Position
number (SEQ ID NO 8) M 4 4 I S 7 7 D L 11 11 N A 19 19 V Q 42 37 L
P 64 59 S G 70 65 S
[0043] With reference to Table 5, in another embodiment, "Class X",
the monoclonal antibodies of the present invention comprise a VL
region having the substitution/backmutation at positions 11 and 19.
In another embodiment "Class Y" the monoclonal antibodies of "Class
X" further comprise the substitution in position 42. In a further
embodiment the monoclonal antibodies of "Class X" or "Class Y"
further comprise a back mutation in positions 7, 64 or 70 to form
"Class Z".
[0044] Again with reference to Table 5, in another embodiment the
monoclonal antibodies of the present invention comprise a VL region
having a substitution at position 4 (corresponding to L11 (SEQ ID
NO.78).
[0045] The following table (Table 6) includes details of seven
different light chain variable (VL) regions which may form part of
the antibodies of the present invention. Each of the disclosed VL
is, or is based on, the L13 VL (SEQ ID NO. 20) optionally further
comprising the substitutions mentioned in the table where the L13
residue at the relevant position is substituted with the 2A10
residue at that position (in the table, "-" means that there is no
substitution in that position, and so the residue remains as in the
sequence of L13):
TABLE-US-00006 TABLE 6 Numerical Residue No. 4 7 11 19 42 64 70
Kabat No. 4 7 11 19 37 59 65 2A10 New VL I D N V L S S (SEQ ID L13
NO. X) M S L A Q P G L11 (78) I -- -- -- -- -- -- L13 (20) -- -- --
-- -- -- -- L14 (21) -- -- -- -- L -- -- L15 (22) -- -- -- -- L S
-- L16 (23) -- -- N V L -- -- L17 (24) -- D -- -- -- S S L18 (25)
-- D N V L S S
[0046] Accordingly there is provided a monoclonal antibody
comprising the light chain variable region having a sequence given
in any one of SEQ ID NOs: 20-25 or 78. In another embodiment there
is provided a monoclonal antibody comprising a light chain having a
sequence given in any one of SEQ ID NOs: 35-40 or 80.
[0047] Alternatively there is provided a monoclonal antibody
comprising the light chain variable region having the sequence
given in SEQ ID NO. 19, or a light chain having the sequence given
in SEQ ID NO. 34.
[0048] In a particular embodiment the antibody comprises the VL
regions provided in SEQ ID NOs 20, 23 and 25 or the light chains
provided in SEQ ID NOs 35, 38 and 40.
Specific Combinations of Heavy and Light Chain Variable Regions
[0049] The antibodies of the present invention comprise a heavy
chain variable region and a light chain variable region.
[0050] In one embodiment the antibody comprises: [0051] (a) A heavy
chain variable region having the amino acid sequence of SEQ ID NO.
77 (H1 variable region) further comprising a number of
substitutions at one or more of positions 12, 20, 38, 40, 48, 67,
68, 70, 72, 74, 76, 79 and 91; wherein each substituted amino acid
residue is replaced with the amino acid residue at the equivalent
position in SEQ ID NO 7 (the heavy chain variable region of the
donor antibody 2A10) and the number of substitutions is between 1
and 13. In other embodiments the number of substitutions is 1, or
2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11, or 12,
or 13; and [0052] (b) a light chain variable region selected from
SEQ ID NOs 20-25 or 78.
[0053] Particular embodiments are antibodies comprising the
following combinations of heavy and light chain variable regions:
H1L13 (SEQ ID 77+SEQ ID 20), H5L13 (SEQ ID 11+SEQ ID 20), H6L13
(SEQ ID 12+SEQ ID 20), H14L13 (SEQ ID 14+SEQ ID 20), H15L13 (SEQ ID
15+SEQ ID 20), H16L13 (SEQ ID 16+SEQ ID 20), H17L13 (SEQ ID 17+SEQ
ID 20), H18L13 (SEQ ID 18+SEQ ID 20), H19L13 (SEQ ID 85+SEQ ID 20),
H20L13 (SEQ ID 86+SEQ ID 20), H21L13 (SEQ ID 87+SEQ ID 20), H22L13
(SEQ ID 88+SEQ ID 20), H23L13 (SEQ ID 89+SEQ ID 20), H24L13 (SEQ ID
90+SEQ ID 20), H25L13 (SEQ ID 91+SEQ ID 20), H700L13 (SEQ ID 13+SEQ
ID 20), H1L16 (SEQ ID 77+SEQ ID 23), H5L16 (SEQ ID 11+SEQ ID 23),
H6L16 (SEQ ID 12+SEQ ID 23), H14L16 (SEQ ID 14+SEQ ID 23), H15L16
(SEQ ID 15+SEQ ID 23), H16L16 (SEQ ID 16+SEQ ID 23), H17L16 (SEQ ID
17+SEQ ID 23), H18L16 (SEQ ID 18+SEQ ID 23), H19L16 (SEQ ID 85+SEQ
ID 23), H20L16 (SEQ ID 86+SEQ ID 23), H21L16 (SEQ ID 87+SEQ ID 23),
H22L16 (SEQ ID 88+SEQ ID 23), H23L16 (SEQ ID 89+SEQ ID 23), H24L16
(SEQ ID 90+SEQ ID 23), H25L16 (SEQ ID 91+SEQ ID 23), H700L16 (SEQ
ID 13+SEQ ID 23), H1L18 (SEQ ID 77+SEQ ID 25), H5L18 (SEQ ID 11+SEQ
ID 25), H6L18 (SEQ ID 12+SEQ ID 25), H14L18 (SEQ ID 14+SEQ ID 25),
H15L18 (SEQ ID 15+SEQ ID 25), H16L18 (SEQ ID 16+SEQ ID 25), H17L18
(SEQ ID 17+SEQ ID 25), H18L18 (SEQ ID 18+SEQ ID 25), H19L18 (SEQ ID
85+SEQ ID 25), H20L18 (SEQ ID 86+SEQ ID 25), H21L18 (SEQ ID 87+SEQ
ID 25), H22L18 (SEQ ID 88+SEQ ID 25), H23L18 (SEQ ID 89+SEQ ID 25),
H24L18 (SEQ ID 90+SEQ ID 25), H25L18 (SEQ ID 91+SEQ ID 25), H700L18
(SEQ ID 13+SEQ ID 25).
[0054] Other embodiments are antibodies comprising the following
combinations of heavy and light chain variable regions: H1L14 (SEQ
ID 77+SEQ ID 21), H5L14 (SEQ ID 11+SEQ ID 21), H6L14 (SEQ ID 12+SEQ
ID 21), H14L14 (SEQ ID 14+SEQ ID 21), H15L14 (SEQ ID 15+SEQ ID 21),
H16L14 (SEQ ID 16+SEQ ID 21), H17L14 (SEQ ID 17+SEQ ID 21), H18L14
(SEQ ID 18+SEQ ID 21), H19L14 (SEQ ID 85+SEQ ID 21), H20L14 (SEQ ID
86+SEQ ID 21), H21L14 (SEQ ID 87+SEQ ID 21), H22L14 (SEQ ID 88+SEQ
ID 21), H23L14 (SEQ ID 89+SEQ ID 21), H24L14 (SEQ ID 90+SEQ ID 21),
H25L14 (SEQ ID 91+SEQ ID 21), H700L14 (SEQ ID 13+SEQ ID 21), H1L15
(SEQ ID 77+SEQ ID 22), H5L15 (SEQ ID 11+SEQ ID 22), H6L15 (SEQ ID
12+SEQ ID 22), H14L15 (SEQ ID 14+SEQ ID 22), H15L15 (SEQ ID 15+SEQ
ID 22), H16L15 (SEQ ID 16+SEQ ID 22), H17L15 (SEQ ID 17+SEQ ID 22),
H18L15 (SEQ ID 18+SEQ ID 22), H19L15 (SEQ ID 85+SEQ ID 22), H20L15
(SEQ ID 86+SEQ ID 22), H21L15 (SEQ ID 87+SEQ ID 22), H22L15 (SEQ ID
88+SEQ ID 22), H23L15 (SEQ ID 89+SEQ ID 22), H24L15 (SEQ ID 90+SEQ
ID 22), H25L15 (SEQ ID 91+SEQ ID 22), H700L15 (SEQ ID 13+SEQ ID
22), H1L17 (SEQ ID 77+SEQ ID 24), H5L17 (SEQ ID 11+SEQ ID 24),
H6L17 (SEQ ID 12+SEQ ID 24), H14L17 (SEQ ID 14+SEQ ID 24), H15L17
(SEQ ID 15+SEQ ID 24), H16L17 (SEQ ID 16+SEQ ID 24), H17L17 (SEQ ID
17+SEQ ID 24), H18L17 (SEQ ID 18+SEQ ID 24), H19L17 (SEQ ID 85+SEQ
ID 24), H20L17 (SEQ ID 86+SEQ ID 24), H21L17 (SEQ ID 87+SEQ ID 24),
H22L17 (SEQ ID 88+SEQ ID 24), H23L17 (SEQ ID 89+SEQ ID 24), H24L17
(SEQ ID 90+SEQ ID 24), H25L17 (SEQ ID 91+SEQ ID 24), H700L17 (SEQ
ID 13+SEQ ID 24).
[0055] Further embodiments are antibodies comprising the following
combinations of heavy and light chain variable regions: H1L6 (SEQ
ID 77+SEQ ID 19), H5L6 (SEQ ID 11+SEQ ID 19), H6L6 (SEQ ID 12+SEQ
ID 19), H14L6 (SEQ ID 14+SEQ ID 19), H15L6 (SEQ ID 15+SEQ ID 19),
H16L6 (SEQ ID 16+SEQ ID 19), H17L6 (SEQ ID 17+SEQ ID 19), H18L6
(SEQ ID 18+SEQ ID 19), H19L6 (SEQ ID 85+SEQ ID 19), H20L6 (SEQ ID
86+SEQ ID 19), H21L6 (SEQ ID 87+SEQ ID 19), H22L6 (SEQ ID 88+SEQ ID
19), H23L6 (SEQ ID 89+SEQ ID 19), H24L6 (SEQ ID 90+SEQ ID 19),
H25L6 (SEQ ID 91+SEQ ID 19), H700L6 (SEQ ID 13+SEQ ID 19), H5L11
(SEQ ID 11+SEQ ID 78), H6L11 (SEQ ID 12+SEQ ID 78), H14L11 (SEQ ID
14+SEQ ID 78), H15L11 (SEQ ID 15+SEQ ID 78), H16L11 (SEQ ID 16+SEQ
ID 78), H17L11 (SEQ ID 17+SEQ ID 78), H18L11 (SEQ ID 18+SEQ ID 78),
H19L11 (SEQ ID 85+SEQ ID 78), H20L11 (SEQ ID 86+SEQ ID 78), H21L11
(SEQ ID 87+SEQ ID 78), H22L11 (SEQ ID 88+SEQ ID 78), H23L11 (SEQ ID
89+SEQ ID 78), H24L11 (SEQ ID 90+SEQ ID 78), H25L11 (SEQ ID 91+SEQ
ID 78), H700L11 (SEQ ID 13+SEQ ID 78).
[0056] Whole Antibodies
[0057] Further, the invention also provides a humanised antibody
which binds to and neutralises NOGO, preferably human NOGO, more
preferably human NOGO-A. More specifically there is provided a
humanised antibody comprising a heavy chain variable region as
described herein and a light chain variable region as described
herein.
[0058] The humanised antibodies of the present invention bind to
human NOGO with a comparable affinity to that of the murine donor
antibody 2A10. In one embodiment the binding of the antibody of the
present invention to NOGO has an affinity constant (KD, as measured
by BiaCore techniques) within 10 fold of 2A10, and in another
embodiment the affinity constant is within three or two fold of
2A10. In another embodiment the affinity constant is within three
or two fold of that of 2A10 and the off-rate (kd) is within 10
fold, or three fold, or two fold of 2A10. The method of measuring
the affinity constant and the off-rate of the antibody should be
clear to the skilled reader, however the kinetic analysis BiaCore
method given in Example 5 of this document is illustrative in this
regard. For example, the affinity constant and the off-rate of 2A10
as measured by BiaCore kinetic analysis is about commonly in the
region of 1 nM and 1.84.times.10.sup.-3 (kd 1/s) respectively; in
the same assay the antibodies of one embodiment of the present
invention will have an affinity constant of less than 8-10 nM and
1.84.times.10.sup.-2.
[0059] In typical embodiments, the antibodies of the invention are
of the IgG class, more typically human IgG1 or IgG4, with a K type
human light chain.
[0060] A further aspect of the invention provides a pharmaceutical
composition comprising an anti-NOGO antibody of the present
invention or functional fragment thereof together with a
pharmaceutically acceptable diluent or carrier.
[0061] In a further aspect, the present invention provides a method
of treatment or prophylaxis of stroke (particularly ischemic
stroke) and other neurological diseases, in particular Alzheimer's
disease, in a human which comprises administering to said human in
need thereof an effective amount of an anti-NOGO antibody of the
invention or functional fragments thereof.
[0062] In another aspect, the invention provides the use of an
anti-NOGO antibody of the invention or a functional fragment
thereof in the preparation of a medicament for treatment or
prophylaxis of stroke (particularly ischemic stroke) and other
neurological diseases, in particular Alzheimer's disease.
[0063] In a further aspect, the present invention provides a method
of inhibiting neurodegeneration and/or promoting functional
recovery in a human patient afflicted with, or at risk of
developing, a stroke (particularly ischemic stroke) or other
neurological disease, in particular Alzheimer's disease, which
comprises administering to said human in need thereof an effective
amount of an anti-NOGO antibody of the invention or a functional
fragment thereof.
[0064] In a yet further aspect, the invention provides the use of
an anti-NOGO antibody of the invention or a functional fragment
thereof in the preparation of a medicament for inhibiting
neurodegeneration and/or promoting functional recovery in a human
patient afflicted with, or at risk of developing, a stroke and
other neurological disease, in particular Alzheimer's disease.
[0065] Other aspects and advantages of the present invention are
described further in the detailed description and the preferred
embodiments thereof.
[0066] In one embodiment the full length antibodies are those
comprising the light chains of SEQ ID NOs 34-40 and the heavy
chains of SEQ ID NOs 92-98; and in particular the light chains of
SEQ ID NOs 35, 38 or 40 and heavy chains of SEQ ID NOs 92, 93, 94
or 98. It will be apparent to those skilled in the art that all of
the sequences given for the full length antibody chains in Table 7
and 12 (and appended sequences) represent the heavy chain or light
chains prior to any processing (e.g. host cell mediated processing)
for removal of a signal sequence. Typically the processed forms of
the antibody chains will begin at position 20 (after the removal of
the signal sequence (residues 1-19) which corresponds to SEQ ID NO.
75). The present invention provides the antibodies having the
polypeptide sequences listed (after removal of the first 19 amino
acids of the signal sequence), and also provides the antibodies in
the form in which they are produced and purified from host cells
expressing the polynucleotides encoding the heavy and light
chain.
[0067] Table 7, Specific full length antibodies include:
TABLE-US-00007 Antibody Light Chain Heavy Chain H5L13 FL SEQ ID NO.
35 SEQ ID NO. 26 H6L13 FL SEQ ID NO. 35 SEQ ID NO. 27 H19L13 FL SEQ
ID NO. 35 SEQ ID NO. 92 H20L13 FL SEQ ID NO. 35 SEQ ID NO. 93
H21L13 FL SEQ ID NO. 35 SEQ ID NO. 94 H25L13 FL SEQ ID NO. 35 SEQ
ID NO. 98 H16L16 FL SEQ ID NO. 38 SEQ ID NO. 31 H19L16 FL SEQ ID
NO. 38 SEQ ID NO. 92 H20L16 FL SEQ ID NO. 38 SEQ ID NO. 93 H21L16
FL SEQ ID NO. 38 SEQ ID NO. 94 H25L16 FL SEQ ID NO. 38 SEQ ID NO.
98 H16L18 FL SEQ ID NO. 40 SEQ ID NO. 31 H18L16 FL SEQ ID NO. 38
SEQ ID NO. 33 H19L18 FL SEQ ID NO. 40 SEQ ID NO. 92 H20L18 FL SEQ
ID NO. 40 SEQ ID NO. 93 H21L18 FL SEQ ID NO. 40 SEQ ID NO. 94
H25L18 FL SEQ ID NO. 40 SEQ ID NO. 98
[0068] Alternatively the substitutions mentioned above, being the
back mutations to the exact amino residue found in the equivalent
position within the donor 2A10 murine sequence, may be any
substitution to an amino acid which is a conservative substitution
of the exact residue found in the equivalent position within the
donor 2A10 murine sequence. The term "conservative substitution" is
clear to the skilled reader, and includes for instance a
substitution of an amino acid with another amino acid residue
having a similar physical, chemical or structural property such as
pH, charge, hydrophobicity, aromaticity etc.
DESCRIPTION OF THE FIGURES
[0069] FIGS. 1 A and B, ELISA data for monoclonal antibody
supernatants binding to recombinant human NOGO.
[0070] FIGS. 2 A and B, ELISA data for purified monoclonal antibody
binding to recombinant human NOGO.
[0071] FIGS. 3 A and B, ELISA data for monoclonal antibody
supernatants binding to recombinant human NOGO.
[0072] FIGS. 4 A and B, ELISA data for monoclonal antibody
supernatants binding to recombinant human NOGO.
[0073] FIGS. 5 A and B, ELISA data for monoclonal antibody
supernatants binding to recombinant human NOGO.
[0074] FIG. 6, A to F, FACS data for purified antibody binding to
human NOGO expressed by human neuroblastoma cells.
[0075] FIG. 7, Competition ELISA results
DETAILED DESCRIPTION OF THE INVENTION
[0076] Antibodies of the invention typically have the structure of
a natural antibody or functional fragment thereof. The antibody may
therefore comprise a full length antibody, a (Fab').sub.2 fragment,
a Fab fragment, a light chain dimer or a heavy chain dimer. The
antibody may be an IgG1, IgG2, IgG3, or IgG4; or IgM; IgA, IgE or
IgD or a modified variant thereof. The constant domain of the
antibody heavy chain may be selected accordingly. The light chain
constant domain may be a kappa or lambda constant domain.
Furthermore, the antibody may comprise modifications of all classes
eg. IgG dimers, Fc mutants that no longer bind Fc receptors or
mediate Clq binding. The antibody may also be a chimeric antibody
of the type described in WO86/01533 which comprises an antigen
binding region and a non-immunoglobulin region. The antigen binding
region is an antibody light chain variable domain or heavy chain
variable domain. Typically the antigen binding region comprises
both light and heavy chain variable domains. The non-immunoglobulin
region is fused at its C terminus to the antigen binding region.
The non-immunoglobulin region is typically a non-immunoglobulin
protein and may be an enzyme, a toxin or protein having known
binding specificity. The two regions of this type of chimeric
antibody may be connected via a cleavable linker sequence.
Immunoadhesins having the CDRs as hereinbefore described are also
contemplated in the present invention.
[0077] The constant region is selected according to the
functionality required. Normally an IgG1 will demonstrate lytic
ability through binding to complement and/or will mediate ADCC
(antibody dependent cell cytotoxicity). An IgG4 will be preferred
if a non-cytotoxic blocking antibody is required. However, IgG4
antibodies can demonstrate instability in production and therefore
it may be more preferable to modify the generally more stable IgG1.
Suggested modifications are described in EP0307434 preferred
modifications include at positions 235 and 237. The invention
therefore provides a lytic or a non-lytic form of an antibody
according to the invention.
[0078] In one embodiment the antibody of the invention is a full
length (i.e. a tetramer comprising two heavy and two light chains)
non-lytic IgG1 antibody having the VH or VL sequences described
supra. In another embodiment we provide a full length non-lytic
IgG1 antibody having the VHs of SEQ ID NOs 11, 12, 16, 18 or 85,
86, 87 or 91; and VLs of SEQ ID NOs 20, 23 or 25.
[0079] In a further aspect, the invention provides polynucleotides
encoding the disclosed heavy or light chains or variable regions.
For example the invention provides polynucleotides encoding VH
having the sequence contained in SEQ ID NOs 45-52, 99-105 and VL
regions having the sequence contained in SEQ ID NOs 53-59.
[0080] "NOGO" refers to any NOGO polypeptide, including variant
forms. This includes, but is not limited to, NOGO-A having 1192
amino acid residues (GenBank accession no. AJ251383); NOGO-B, a
splice variant which lacks residues 186 to 1004 in the putative
extracellular domain (GenBank accession no. AJ251384) and a shorter
splice variant, NOGO-C, which also lacks residues 186 to 1004 and
also has smaller, alternative amino terminal domain (GenBank
accession no. AJ251385) (Prinjha et al (2000) supra). All
references to "NOGO" herein is understood to include any and all
variant forms of NOGO such as NOGO-A and the splice variants
described, unless a specific form is indicated.
[0081] "Neutralising" and grammatical variations thereof refers to
inhibition, either total or partial, of NOGO function including its
binding to neurones and inhibition of neurite growth.
[0082] "Altered antibody" refers to a protein encoded by an altered
immunoglobulin coding region, which may be obtained by expression
in a selected host cell. Such altered antibodies include engineered
antibodies (e.g., chimeric, reshaped, humanized or vectored
antibodies) or antibody fragments lacking all or part of an
immunoglobulin constant region, e.g., Fv, Fab, or F(ab).sub.2 and
the like.
[0083] "Altered immunoglobulin coding region" refers to a nucleic
acid sequence encoding altered antibody. When the altered antibody
is a CDR-grafted or humanized antibody, the sequences that encode
the complementarity determining regions (CDRs) from a non-human
immunoglobulin are inserted into a first immunoglobulin partner
comprising human variable framework sequences. Optionally, the
first immunoglobulin partner is operatively linked to a second
immunoglobulin partner.
[0084] The terms Fv, Fc, Fd, Fab, or F(ab).sub.2 are used with
their standard meanings (see, e.g., Harlow et al., Antibodies A
Laboratory Manual, Cold Spring Harbor Laboratory, (1988)).
[0085] As used herein, an "engineered antibody" describes a type of
altered antibody, i.e., a full-length synthetic antibody (e.g., a
chimeric, reshaped or humanized antibody as opposed to an antibody
fragment) in which a portion of the light and/or heavy chain
variable domains of a selected acceptor antibody are replaced by
analogous parts from one or more donor antibodies which have
specificity for the selected epitope. For example, such molecules
may include antibodies characterized by a humanized heavy chain
associated with an unmodified light chain (or chimeric light
chain), or vice versa. Engineered antibodies may also be
characterized by alteration of the nucleic acid sequences encoding
the acceptor antibody light and/or heavy variable domain framework
regions in order to retain donor antibody binding specificity.
These antibodies can comprise replacement of one or more CDRs
(preferably all) from the acceptor antibody with CDRs from a donor
antibody described herein.
[0086] A "chimeric antibody" refers to a type of engineered
antibody which contains a naturally-occurring variable region
(light chain and heavy chains) derived from a donor antibody in
association with light and heavy chain constant regions derived
from an acceptor antibody.
[0087] A "humanized antibody" refers to a type of engineered
antibody having its CDRs derived from a non-human donor
immunoglobulin, the remaining immunoglobulin-derived parts of the
molecule being derived from one (or more) human immunoglobulin(s).
In addition, framework support residues may be altered to preserve
binding affinity (see, e.g., Queen et al., Proc. Natl. Acad Sci
USA, 86:10029-10032 (1989), Hodgson et al., Bio/Technology, 9:421
(1991)). A suitable human acceptor antibody may be one selected
from a conventional database, e.g., the KABAT.RTM. database, Los
Alamos database, and Swiss Protein database, by homology to the
nucleotide and amino acid sequences of the donor antibody. A human
antibody characterized by a homology to the framework regions of
the donor antibody (on an amino acid basis) may be suitable to
provide a heavy chain constant region and/or a heavy chain variable
framework region for insertion of the donor CDRs. A suitable
acceptor antibody capable of donating light chain constant or
variable framework regions may be selected in a similar manner. It
should be noted that the acceptor antibody heavy and light chains
are not required to originate from the same acceptor antibody. The
prior art describes several ways of producing such humanised
antibodies--see for example EP-A-0239400 and EP-A-054951
[0088] The term "donor antibody" refers to an antibody (monoclonal,
and/or recombinant) which contributes the amino acid sequences of
its variable regions, CDRs, or other functional fragments or
analogs thereof to a first immunoglobulin partner, so as to provide
the altered immunoglobulin coding region and resulting expressed
altered antibody with the antigenic specificity and neutralizing
activity characteristic of the donor antibody.
[0089] The term "acceptor antibody" refers to an antibody
(monoclonal and/or recombinant) heterologous to the donor antibody,
which contributes all (or any portion, but preferably all) of the
amino acid sequences encoding its heavy and/or light chain
framework regions and/or its heavy and/or light chain constant
regions to the first immunoglobulin partner. Preferably a human
antibody is the acceptor antibody.
[0090] "CDRs" are defined as the complementarity determining region
amino acid sequences of an antibody which are the hypervariable
regions of immunoglobulin heavy and light chains. See, e.g., Kabat
et al., Sequences of Proteins of Immunological Interest, 4th Ed.,
U.S. Department of Health and Human Services, National Institutes
of Health (1987). There are three heavy chain and three light chain
CDRs (or CDR regions) in the variable portion of an immunoglobulin.
Thus, "CDRs" as used herein refers to all three heavy chain CDRs,
or all three light chain CDRs (or both all heavy and all light
chain CDRs, if appropriate). The structure and protein folding of
the antibody may mean that other residues are considered part of
the antigen binding region and would be understood to be so by a
skilled person. See for example Chothia et al., (1989)
Conformations of immunoglobulin hypervariable regions; Nature 342,
p 877-883.
[0091] CDRs provide the majority of contact residues for the
binding of the antibody to the antigen or epitope. CDRs of interest
in this invention are derived from donor antibody variable heavy
and light chain sequences, and include analogs of the naturally
occurring CDRs, which analogs also share or retain the same antigen
binding specificity and/or neutralizing ability as the donor
antibody from which they were derived.
[0092] A "functional fragment" is a partial heavy or light chain
variable sequence (e.g., minor deletions at the amino or carboxy
terminus of the immunoglobulin variable region) which retains the
same antigen binding specificity and the same or similar
neutralizing ability as the antibody from which the fragment was
derived.
[0093] An "analog" is an amino acid sequence modified by at least
one amino acid, wherein said modification can be chemical or a
substitution or a rearrangement of a few amino acids (i.e., no more
than 10), which modification permits the amino acid sequence to
retain the biological characteristics, e.g., antigen specificity
and high affinity, of the unmodified sequence.
[0094] Analogs may also arise as allelic variations. An "allelic
variation or modification" is an alteration in the nucleic acid
sequence. Such variations or modifications may be due to degeneracy
in the genetic code or may be deliberately engineered to provide
desired characteristics. These variations or modifications may or
may not result in alterations in any encoded amino acid
sequence.
[0095] The present invention also includes the use of Fab fragments
or F(ab').sub.2 fragments derived from mAbs of the present
invention directed against NOGO. A Fab fragment contains the entire
light chain and amino terminal portion of the heavy chain; and an
F(ab').sub.2 fragment is the fragment formed by two Fab fragments
bound by disulfide bonds. Fab fragments and F(ab').sub.2 fragments
can be obtained by conventional means, e.g., cleavage of mAb with
the appropriate proteolytic enzymes, papain and/or pepsin, or by
recombinant methods. The Fab and F(ab').sub.2 fragments are useful
themselves as therapeutic or prophylactic, and as donors of
sequences including the variable regions and CDR sequences useful
in the formation of recombinant or humanized antibodies as
described herein.
[0096] Altered immunoglobulin molecules can encode altered
antibodies which include engineered antibodies such as chimeric
antibodies and humanized antibodies. A desired altered
immunoglobulin coding region contains CDR-encoding regions that
encode peptides having the antigen specificity of an anti-NOGO
antibody, preferably a high affinity antibody, inserted into a
first immunoglobulin partner (a human framework or human
immunoglobulin variable region).
[0097] Preferably, the first immunoglobulin partner is operatively
linked to a second immunoglobulin partner. The second
immunoglobulin partner is defined above, and may include a sequence
encoding a second antibody region of interest, for example an Fc
region. Second immunoglobulin partners may also include sequences
encoding another immunoglobulin to which the light or heavy chain
constant region is fused in frame or by means of a linker sequence.
Engineered antibodies directed against functional fragments or
analogs of NOGO may be designed to elicit enhanced binding.
[0098] The second immunoglobulin partner may also be associated
with effector agents as defined above, including non-protein
carrier molecules, to which the second immunoglobulin partner may
be operatively linked by conventional means.
[0099] Fusion or linkage between the second immunoglobulin
partners, e.g., antibody sequences, and the effector agent may be
by any suitable means, e.g., by conventional covalent or ionic
bonds, protein fusions, or hetero-bifunctional cross-linkers, e.g.,
carbodiimide, glutaraldehyde, and the like. Such techniques are
known in the art and readily described in conventional chemistry
and biochemistry texts.
[0100] Additionally, conventional linker sequences which simply
provide for a desired amount of space between the second
immunoglobulin partner and the effector agent may also be
constructed into the altered immunoglobulin coding region. The
design of such linkers is well known to those of skill in the art.
In further aspects of the invention we provide diabodies (bivalent
or bispecific), triabodies, tetrabodies and other multivalent scFV
protein species having one or more CDRs as described supra that
bind to and neutralise NOGO function.
[0101] In still a further embodiment, the antibody of the invention
may have attached to it an additional agent. For example, the
procedure of recombinant DNA technology may be used to produce an
engineered antibody of the invention in which the Fc fragment or
CH2-CH3 domain of a full length antibody molecule has been replaced
by an enzyme or other detectable molecule (i.e., a polypeptide
effector or reporter molecule).
[0102] The second immunoglobulin partner may also be operatively
linked to a non-immunoglobulin peptide, protein or fragment thereof
heterologous to the CDR-containing sequence having the antigen
specificity of anti-NOGO antibody. The resulting protein may
exhibit both anti-NOGO antigen specificity and characteristics of
the non-immunoglobulin upon expression. That fusion partner
characteristic may be, e.g., a functional characteristic such as
another binding or receptor domain, or a therapeutic characteristic
if the fusion partner is itself a therapeutic protein, or
additional antigenic characteristics.
[0103] Another desirable protein of this invention may comprise a
full length antibody molecule, having full length heavy and light
chains, or any discrete fragment thereof, such as the Fab or
F(ab').sub.2 fragments, a heavy chain dimer, or any minimal
recombinant fragments thereof such as an Fv or a single-chain
antibody (SCA) or any other molecule with the same specificity as
the selected donor mAb. Such protein may be used in the form of an
altered antibody, or may be used in its unfused form.
[0104] Whenever the second immunoglobulin partner is derived from
an antibody different from the donor antibody, e.g. any isotype or
class of immunoglobulin framework or constant regions, an
engineered antibody results. Engineered antibodies can comprise
immunoglobulin (Ig) constant regions and variable framework regions
from one source, e.g., the acceptor antibody, and one or more
(preferably all) CDRs from the donor antibody. In addition,
alterations, e.g., deletions, substitutions, or additions, of the
acceptor mAb light and/or heavy variable domain framework region at
the nucleic acid or amino acid levels, or the donor CDR regions may
be made in order to retain donor antibody antigen binding
specificity.
[0105] Such engineered antibodies are designed to employ one (or
both) of the variable heavy and/or light chains of the anti-NOGO
mAb or one or more of the heavy or light chain CDRs. The engineered
antibodies may be neutralising, as above defined.
[0106] In addition, the constant region may be altered to enhance
or decrease selective properties of the molecules of the instant
invention. For example, dimerization, binding to Fc receptors, or
the ability to bind and activate complement (see, e.g., Angal et
al., Mol. Immunol, 30:105-108 (1993), Xu et al., J. Biol. Chem,
269:3469-3474 (1994), Winter et al., EP 307,434-B).
[0107] The antibodies of the present invention may be produced by
making a conventional expression vector or recombinant plasmid by
placing these coding sequences for the antibody in operative
association with conventional regulatory control sequences capable
of controlling the replication and expression in, and/or secretion
from, a host cell. Regulatory sequences include promoter sequences,
e.g., CMV promoter, and signal sequences, which can be derived from
other known antibodies. Similarly, a second expression vector can
be produced having a DNA sequence which encodes a complementary
antibody light or heavy chain. Preferably this second expression
vector is identical to the first except insofar as the coding
sequences and selectable markers are concerned, so to ensure as far
as possible that each polypeptide chain is functionally expressed.
Alternatively, the heavy and light chain coding sequences for the
altered antibody may reside on a single vector.
[0108] A selected host cell is co-transfected by conventional
techniques with both the first and second vectors (or simply
transfected by a single vector) to create the transfected host cell
of the invention comprising both the recombinant or synthetic light
and heavy chains. The transfected cell is then cultured by
conventional techniques to produce the engineered antibody of the
invention. The antibody which includes the association of both the
recombinant heavy chain and/or light chain is screened from culture
by appropriate assay, such as ELISA or RIA. Similar conventional
techniques may be employed to construct other altered antibodies
and molecules.
[0109] Suitable vectors for the cloning and subcloning steps
employed in the methods and construction of the compositions of
this invention may be selected by one of skill in the art. For
example, the conventional pUC series of cloning vectors may be
used. One vector, pUC19, is commercially available from supply
houses, such as Amersham (Buckinghamshire, United Kingdom) or
Pharmacia (Uppsala, Sweden). Additionally, any vector which is
capable of replicating readily, has an abundance of cloning sites
and selectable genes (e.g., antibiotic resistance), and is easily
manipulated may be used for cloning. Thus, the selection of the
cloning vector is not a limiting factor in this invention.
[0110] Similarly, the vectors employed for expression of the
antibodies may be selected by one of skill in the art from any
conventional vector. The vectors also contain selected regulatory
sequences (such as CMV or RSV promoters) which direct the
replication and expression of heterologous DNA sequences in
selected host cells. These vectors contain the above described DNA
sequences which code for the antibody or altered immunoglobulin
coding region. In addition, the vectors may incorporate the
selected immunoglobulin sequences modified by the insertion of
desirable restriction sites for ready manipulation.
[0111] The expression vectors may also be characterized by genes
suitable for amplifying expression of the heterologous DNA
sequences, e.g., the mammalian dihydrofolate reductase gene (DHFR).
Other preferable vector sequences include a poly A signal sequence,
such as from bovine growth hormone (BGH) and the betaglobin
promoter sequence (betaglopro). The expression vectors useful
herein may be synthesized by techniques well known to those skilled
in this art.
[0112] The components of such vectors, e.g. replicons, selection
genes, enhancers, promoters, signal sequences and the like, may be
obtained from commercial or natural sources or synthesized by known
procedures for use in directing the expression and/or secretion of
the product of the recombinant DNA in a selected host. Other
appropriate expression vectors of which numerous types are known in
the art for mammalian, bacterial, insect, yeast, and fungal
expression may also be selected for this purpose.
[0113] The present invention also encompasses a cell line
transfected with a recombinant plasmid containing the coding
sequences of the antibodies or altered immunoglobulin molecules
thereof. Host cells useful for the cloning and other manipulations
of these cloning vectors are also conventional. However, most
desirably, cells from various strains of E. coli are used for
replication of the cloning vectors and other steps in the
construction of altered antibodies of this invention.
[0114] Suitable host cells or cell lines for the expression of the
antibody of the invention are preferably mammalian cells such as
NSO, Sp2/0, CHO (e.g. DG44), COS, a fibroblast cell (e.g., 3T3),
and myeloma cells, and more preferably a CHO or a myeloma cell.
Human cells may be used, thus enabling the molecule to be modified
with human glycosylation patterns. Alternatively, other eukaryotic
cell lines may be employed. The selection of suitable mammalian
host cells and methods for transformation, culture, amplification,
screening and product production and purification are known in the
art. See, e.g., Sambrook et al., cited above.
[0115] Bacterial cells may prove useful as host cells suitable for
the expression of the recombinant Fabs of the present invention
(see, e.g., Pluckthun, A., Immunol. Rev., 130:151-188 (1992)).
However, due to the tendency of proteins expressed in bacterial
cells to be in an unfolded or improperly folded form or in a
non-glycosylated form, any recombinant Fab produced in a bacterial
cell would have to be screened for retention of antigen binding
ability. If the molecule expressed by the bacterial cell was
produced in a properly folded form, that bacterial cell would be a
desirable host. For example, various strains of E. coli used for
expression are well-known as host cells in the field of
biotechnology. Various strains of B. subtilis, Streptomyces, other
bacilli and the like may also be employed in this method.
[0116] Where desired, strains of yeast cells known to those skilled
in the art are also available as host cells, as well as insect
cells, e.g. Drosophila and Lepidoptera and viral expression
systems. See, e.g. Miller et al., Genetic Engineering, 8:277-298,
Plenum Press (1986) and references cited therein.
[0117] The general methods by which the vectors may be constructed,
the transfection methods required to produce the host cells of the
invention, and culture methods necessary to produce the antibody of
the invention from such host cell are all conventional techniques.
Typically, the culture method of the present invention is a
serum-free culture method, usually by culturing cells serum-free in
suspension. Likewise, once produced, the antibodies of the
invention may be purified from the cell culture contents according
to standard procedures of the art, including ammonium sulfate
precipitation, affinity columns, column chromatography, gel
electrophoresis and the like. Such techniques are within the skill
of the art and do not limit this invention. For example,
preparation of altered antibodies are described in WO 99/58679 and
WO 96/16990.
[0118] Yet another method of expression of the antibodies may
utilize expression in a transgenic animal, such as described in
U.S. Pat. No. 4,873,316. This relates to an expression system using
the animal's casein promoter which when transgenically incorporated
into a mammal permits the female to produce the desired recombinant
protein in its milk.
[0119] In a further aspect of the invention there is provided a
method of producing an antibody of the invention which method
comprises the step of culturing a host cell transformed or
transfected with a vector encoding the light and/or heavy chain of
the antibody of the invention and recovering the antibody thereby
produced.
[0120] In accordance with the present invention there is provided a
method of producing an anti-NOGO antibody which specifically binds
to and neutralises the activity of human NOGO-A which method
comprises the steps of; [0121] (a) providing a first vector
encoding a heavy chain of the antibody; [0122] (b) providing a
second vector encoding the light chain of the antibody; [0123] (c)
tranforming a mammalian host cell (e.g. CHO) with said first and
second vectors; [0124] (d) culturing the host cell of step (c)
under conditions conducive to the secretion of the antibody from
said host cell into said culture media; [0125] (e) recovering the
secreted antibody of step (d).
[0126] Once expressed by the desired method, the antibody is then
examined for in vitro activity by use of an appropriate assay.
Presently conventional ELISA assay formats are employed to assess
qualitative and quantitative binding of the antibody to NOGO.
Additionally, other in vitro assays may also be used to verify
neutralizing efficacy prior to subsequent human clinical studies
performed to evaluate the persistence of the antibody in the body
despite the usual clearance mechanisms.
[0127] The therapeutic agents of this invention may be administered
as a prophylactic or following the stroke event/on-set of clinical
symptoms, or as otherwise needed. The dose and duration of
treatment relates to the relative duration of the molecules of the
present invention in the human circulation, and can be adjusted by
one of skill in the art depending upon the condition being treated
and the general health of the patient. It is envisaged that
repeated dosing (e.g. once a week or once every two weeks) over an
extended time period (e.g. four to six months) maybe required to
achieve maximal therapeutic efficacy.
[0128] The mode of administration of the therapeutic agent of the
invention may be any suitable route which delivers the agent to the
host. The antagonists and antibodies, and pharmaceutical
compositions of the invention are particularly useful for
parenteral administration, i.e., subcutaneously, intrathecally,
intraperitoneally, intramuscularly, intravenously, or intranasally,
of which intravenously is particularly preferred.
[0129] Therapeutic agents of the invention may be prepared as
pharmaceutical compositions containing an effective amount of the
antagonist or antibody of the invention as an active ingredient in
a pharmaceutically acceptable carrier. In the prophylactic agent of
the invention, an aqueous suspension or solution containing the
engineered antibody, preferably buffered at physiological pH, in a
form ready for injection is preferred. The compositions for
parenteral administration will commonly comprise a solution of the
antagonist or antibody of the invention or a cocktail thereof
dissolved in a pharmaceutically acceptable carrier, preferably an
aqueous carrier. A variety of aqueous carriers may be employed,
e.g., 0.9% saline, 0.3% glycine, and the like. These solutions are
sterile and generally free of particulate matter. These solutions
may be sterilized by conventional, well known sterilization
techniques (e.g., filtration). The compositions may contain
pharmaceutically acceptable auxiliary substances as required to
approximate physiological conditions such as pH adjusting and
buffering agents, etc. The concentration of the antagonist or
antibody of the invention in such pharmaceutical formulation can
vary widely, i.e., from less than about 0.5%, usually at or at
least about 1% to as much as 15 or 20% by weight and will be
selected primarily based on fluid volumes, viscosities, etc.,
according to the particular mode of administration selected.
[0130] Thus, a pharmaceutical composition of the invention for
intramuscular injection could be prepared to contain 1 mL sterile
buffered water, and between about 1 ng to about 100 mg, e.g. about
50 ng to about 30 mg or more preferably, about 5 mg to about 25 mg,
of an antagonist or antibody of the invention. Similarly, a
pharmaceutical composition of the invention for intravenous
infusion could be made up to contain about 250 ml of sterile
Ringer's solution, and about 1 to about 30 and preferably 5 mg to
about 25 mg of an engineered antibody of the invention. Actual
methods for preparing parenterally administrable compositions are
well known or will be apparent to those skilled in the art and are
described in more detail in, for example, Remington's
Pharmaceutical Science, 15th ed., Mack Publishing Company, Easton,
Pa. For the preparation of intravenously administrable antibody
formulations of the invention see Lasmar U and Parkins D "The
formulation of Biopharmaceutical products", Pharma. Sci. Tech.
today, page 129-137, Vol. 3 (3 Apr. 2000), Wang, W "Instability,
stabilisation and formulation of liquid protein pharmaceuticals",
Int. J. Pharm 185 (1999) 129-188, Stability of Protein
Pharmaceuticals Part A and B ed Ahern T. J., Manning M. C., New
York, N.Y.: Plenum Press (1992), Akers, M. J. "Excipient-Drug
interactions in Parenteral Formulations", J. Pharm Sci 91 (2002)
2283-2300, Imamura, K et al "Effects of types of sugar on
stabilization of Protein in the dried state", J Pharm Sci 92 (2003)
266-274,Izutsu, Kkojima, S. "Excipient crystallinity and its
protein-structure-stabilizing effect during freeze-drying", J.
Pharm. Pharmacol, 54 (2002) 1033-1039, Johnson, R,
"Mannitol-sucrose mixtures-versatile formulations for protein
lyophilization", J. Pharm. Sci, 91 (2002) 914-922.
[0131] Ha,E Wang W, Wang Y. j. "Peroxide formation in polysorbate
80 and protein stability", J. Pharm Sci, 91, 2252-2264, (2002) the
entire contents of which are incorporated herein by reference and
to which the reader is specifically referred.
[0132] It is preferred that the therapeutic agent of the invention,
when in a pharmaceutical preparation, be present in unit dose
forms. The appropriate therapeutically effective dose can be
determined readily by those of skill in the art. To effectively
treat stroke and other neurological diseases in a human, one dose
of up to 700 mg per 70 kg body weight of an antagonist or antibody
of this invention should be administered parenterally, preferably
i.v. or i.m. (intramuscularly). Such dose may, if necessary, be
repeated at appropriate time intervals selected as appropriate by a
physician. As disclosed in the examples, the present inventors have
been able to demonstrate a positive effect on functional recovery
in the rat model therein when antibodies of the invention were
administered intravenously.
[0133] The antibodies described herein can be lyophilized for
storage and reconstituted in a suitable carrier prior to use. This
technique has been shown to be effective with conventional
immunoglobulins and art-known lyophilization and reconstitution
techniques can be employed.
[0134] Antibodies of the invention may also be used in combination
(i.e. simultaneously, sequentially or separately) with a
neurotrophic factor such as nerve growth factor (NGF), for example
brain derived neurotrophic factor (BDNF), anti-inflammatory agents
such as corticosteroids, and/or tPA. Combinations of a NOGO
antibody of the invention and e.g. tPA maybe assessed in the MCAO
model set forth in the examples below.
[0135] In another aspect, the invention provides a pharmaceutical
composition comprising anti-NOGO antibody of the present invention
or a functional fragment thereof and a pharmaceutically acceptable
carrier for treatment or prophylaxis of stroke and other
neurological diseases.
[0136] In a yet further aspect, the invention provides a
pharmaceutical composition comprising the anti-NOGO antibody of the
present invention or a functional fragment thereof and a
pharmaceutically acceptable carrier for inhibiting
neurodegeneration and/or promoting functional recovery in a human
patient suffering, or at risk of developing, a stroke or other
neurological disease.
[0137] The invention further provides a method of treatment or
prophylaxis of stroke (particularly ischemic stroke) and other
neurological diseases/disorders, in particular Alzheimer's disease,
in a human which comprises administering to said human in need
thereof an effective amount of an anti-NOGO antibody or a
functional fragment thereof. Antibodies of the invention may be
used in methods of treatment to slow or halt the progression and/or
onset of Alzheimer's disease in addition to (or as an alternative
to) treating established disease in a human patient.
[0138] Further the invention provides the use of an anti-NOGO
antibody, or a functional fragment thereof, in the preparation of a
medicament for treatment or prophylaxis of stroke and other
neurological diseases/disorders, in particular Alzheimer's
disease.
[0139] The invention also provides a method of inhibiting
neurodegeneration and/or promoting functional recovery in a human
patient suffering, or at risk of developing, a stroke or other
neurological disease/disorder, in particular Alzheimer's disease,
which comprises administering to said human in need thereof an
effective amount of an anti-NOGO antibody or a functional fragment
thereof.
[0140] In addition the invention provides the use of an anti-NOGO
antibody or a functional fragment thereof in the preparation of a
medicament for inhibiting neurodegeneration and/or promoting
functional recovery in a human patient afflicted with, or at risk
of developing, a stroke and other neurological disease/disorder, in
particular Alzheimer's disease.
[0141] The invention further provides a method of treating or
prophylaxis of stroke or other neurological disease/disorder, in
particular Alzheimer's disease, in a human comprising the step of
parenteral administration of a therapeutically effective amount of
an anti-NOGO antibody. Preferably the anti-NOGO antibody is
administered intravenously.
[0142] Neurological diseases or disorders as used hereinabove
includes, but is not limited to traumatic brain injury, spinal cord
injury, fronto-temporal dementias (tauopathies), peripheral
neuropathy, Parkinson's disease, Huntington's disease, multiple
sclerosis and in particular Alzheimer's disease.
[0143] The invention also provides a method of promoting axonal
sprouting comprising the step of contacting a human axon with an
anti-NOGO antibody. This method may be performed in-vitro or
in-vivo, preferably the method is performed in-vivo.
[0144] In a further aspect therefore there is provided the use of
an anti-NOGO antibody or functional fragment thereof of the
invention comprising CDR's of table 1 and 2; CDR's of Table 3 and
4; or CDR's of table 5 and 6 in intravenously administrable form in
the manufacture of a medicament for the treatment of stroke
(particularly ischemic stroke), brain injury, spinal cord injury,
fronto-temporal dementias (tauopathies), peripheral neuropathy,
Parkinson's disease, Huntington's disease, multiple sclerosis and
in particular Alzheimer's disease in a human patient.
[0145] In a further aspect therefore there is provided a method of
treating stroke (particularly ischemic stroke), brain injury,
spinal cord injury, fronto-temporal dementias (tauopathies),
peripheral neuropathy, Parkinson's disease, Huntington's disease,
multiple sclerosis and in particular Alzheimer's disease in a human
patient which method comprises the intravenous administration of a
therapeutically effective amount of an anti-NOGO antibody of the
invention.
[0146] In a further aspect of the present invention there is
provided a method of promoting axon sprouting of neurons within the
central nervous system of a human subject (e.g. patient) which
method comprises administering (e.g. intravenously administering) a
therapeutically effective amount of an anti-NOGO antibody (e.g. an
anti-NOGO antibody comprising CDRs as set forth herein).
[0147] In a further aspect of the present invention there is
provided the use of an anti-NOGO antibody (e.g. an anti-NOGO
antibody comprising the CDRs set forth herein) in the manufacture
of an intravenously administrable medicament for the treatment of
stroke (particularly ischemic stroke), brain injury, spinal cord
injury, fronto-temporal dementias (tauopathies), peripheral
neuropathy, Parkinson's disease, Huntington's disease, multiple
sclerosis and in particular Alzheimer's disease in a human
patient.
[0148] In a further aspect of the invention there is provided a
method of regenerating axon processes in neurons of the central
nervous system in a human patient afflicted with (or susceptible
to) stroke (particularly ischemic stroke), brain injury, spinal
cord injury, fronto-temporal dementias (tauopathies), peripheral
neuropathy, Parkinson's disease, Huntington's disease, multiple
sclerosis and in particular Alzheimer's disease which method
comprises the step of administering (e.g. intravenously) a
therapeutically effective amount of an anti-NOGO antibody (e.g. an
anti-NOGO antibody having the CDRs set forth herein).
[0149] In a further aspect of the invention there is provided the
use of an anti-NOGO antibody (e.g. an anti-NOGO antibody having the
CDRs set forth herein) in the manufacture of an intravenously
administrable pharmaceutical composition for regenerating axon
processes in neurons of the central nervous system in a human
patient afflicted with (or susceptible to) stroke (particularly
ischemic stroke), brain injury, spinal cord injury, fronto-temporal
dementias (tauopathies), peripheral neuropathy, Parkinson's
disease, Huntington's disease, multiple sclerosis and in particular
Alzheimer's disease.
[0150] In a further aspect of the invention there is provided a
method of modulating the production of an amyloidogenic peptide
comprising contacting a cell which is expressing the precursor from
which the amyloidogenic peptide is derived and a NOGO polypeptide
(e.g. human NOGO-A) with an anti-NOGO antibody (e.g. an anti-NOGO
antibody comprising the CDRs set forth herein, particularly 2A10
and fully human or humanised versions thereof). In typical
embodiments, the precursor is APP. In further typical embodiments
the amyloidogenic peptide is A13, most preferably A1340, A1342 or a
combination of both.
[0151] As used herein, the term "functional recovery" refers to a
motor and/or sensory and/or behavioural improvement in a subject
following e.g. an ischemic event or injury or on-set of clinical
symptoms. Functional recovery in humans may be evaluated by
instruments designed to measure elemental neurological functions
such as motor strength, sensation and coordination, cognitive
functions such as memory, language and the ability to follow
directions, and functional capacities such as basic activities of
daily living or instrumental activities. Recovery of elemental
neurological function can be measured with instruments such as the
NIH Stroke Scale (NIHSS), recovery of cognitive function can be
measured with neuropsychological tests such as Boston Naming Test,
Trail-making Tests, and California Verbal Learning Test, and
activities of daily living may be measured with instruments such as
the ADCS/ADL (Alzheimer's Disease Clinical Studies/Activities of
Daily Living) scale or the Bristol Activities of Daily Living
Scale, all tests and scales known in the art.
Example 1
Construction and Expression of Humanised Anti-NOGO Antibodies
[0152] Murine and Humanised V.sub.H and V.sub.L constructs were
prepared de novo by build up of overlapping oligonucleotides
including restriction sites for cloning into Rld and Rln mammalian
expression vectors as well as a human signal sequence. Hind III and
Spe I restriction sites were introduced to frame the V.sub.H domain
containing the CAMPATH-1H signal sequence for cloning into Rld
containing the human .gamma.1 mutated constant region. Hind III and
BsiWI restriction sites were introduced to frame the V.sub.L domain
containing the CAMPATH-1H signal sequence for cloning into Rln
containing the human kappa constant region.
CAMPATH-1H signal sequence: MGWSCIILFLVATATGVHS (SEQ.I.D.NO:82)
[0153] In parallel a chimeric version of 11C7 was generated (see
WO04/052932). The variable heavy domain sequence (derived from
WO04/052932 Seq ID 43) and variable light domain sequence (derived
from WO04/052932 Seq ID 44) were prepared de novo by build up of
overlapping oligonucleotides. Hind III and SpeI restriction sites
were introduced to frame the V.sub.H domain for cloning into Rld
containing the human .gamma.1 mutated constant region. HindIII and
BsiWI restriction sites were introduced to frame the V.sub.L domain
for cloning into Rln containing the human kappa constant
region.
Example 2
Antibody Expression in CHO Cells
[0154] Rld and Rln plasmids encoding the heavy and light chains
respectively were transiently co-transfected into CHO cells and
expressed at small scale or large scale to produce antibody.
Alternatively the same plasmids were co-transfected into CHO cells
by electroporation and a stable polyclonal population of cells
expressing the appropriate antibody were selected using a
nucleoside-free media. Recombinant antibody was recovered and
purified by affinity chromatography on Protein A sepharose.
Example 3
Humanised Anti-NOGO Antibody Binds to NOGO
[0155] GST-human NOGO-A56 (SEQ ID: 76) at 1 .mu.g/ml in PBS was
coated onto Nunc Immunosorp plates (100 .mu.l per well) at
4.degree. C. overnight. Wells were rinsed once with TBS+0.05% Tween
(TBST) then incubated with 2% BSA in TBST to block non-specific
binding sites at room temperature for 1 hour. Antibodies were
diluted in TBST+2% BSA to 10 .mu.g/ml and 1/2dilutions made from
this. Antibodies were added to wells in duplicate and incubated at
room temperature for 1 hour. Wells were washed three times with
TBST then incubated with anti-human kappa peroxidase conjugate
(1:2000) for 1 hour. The wells were washed three times with TBST
and then incubated with 100 .mu.l OPD peroxidase substrate (Sigma)
per well for 10 minutes. The colour reaction was stopped by the
addition of 25 .mu.l concentrated H.sub.2SO.sub.4. Optical density
at 490 nm was measured using a plate reader. Background values read
from wells with no antibody were subtracted.
[0156] FIGS. 1-5 illustrate the dose-dependent binding of humanised
antibodies in comparison with the chimera (termed HcLc which is the
chimera of 2A10 (comprising the 2A10 murine VH (SEQ ID NO. 7) and
VL (SEQ ID NO:8) and human IgG constant regions)) to human NOGO-A56
(see Example 6 for details) in an ELISA assay. The Y-axis shows the
measured optical density (OD) at 490 nm, a quantitative measure of
antibody captured in the wells. The X-axis shows the concentration
of antibody used (mcg/ml) per well at each data point.
[0157] The antibody material used in FIGS. 1, 3, 4 and 5 was
generated from small scale transient transfections. Human IgG
levels in the supernatant are quantified by ELISA (Example 4). For
FIG. 2, the material used is purified antibody generated by either
the polyclonal expression system or large scale transient
transfections. In these cases, IgG levels were quantified by ELISA
and optical density.
[0158] In another experiment, antibody material was generated from
small scale transient transfections (in triplicate) for the
following humanised antibodies: H16L16, H17L16, H18L16, H16L18 and
the chimeric antibody HcLc. The results from this experiment are
consistent with the data presented in FIGS. 1-5 with the exception
of H17L16 which performed less well than shown in FIG. 1A and FIG.
2. Whilst this observation cannot be explained it should be noted
that the conclusions are contradicted by another experiment with
supernatant material (see FIG. 1A) and an experiment using purified
H17L16 material (FIG. 2), both experiments indicated that H17L16
shows comparable binding to the other optimised variants.
Example 4
Antibody Quantification Protocol
[0159] Nunc Immunosorp plates were coated with capture antibody H19
(goat anti-human IgG chain, Sigma #13382) at 2 .mu.g/ml in
Bicarbonate buffer (Sigma #C3041) and incubated overnight at
4.degree. C. The plates were washed twice with TBS containing 0.05%
Tween20 (TBST) and blocked with 200 .mu.l TBST containing 2% BSA
(block buffer) for 1 hr at room temperature. The plates were washed
twice with TBST. Tissue culture supernatants containing antibody
were titrated across the plate in 2-fold dilution steps into block
buffer and incubated at room temperature for 1 hr. The plates were
washed three times with TBST. HRP conjugated antibody H23 (goat
anti-human kappa chain, Sigma #A7164) was diluted 1:2000 in TBST
and 100 .mu.l added to each well. The plates were incubated at room
temperature for 1 hr. The plates were washed three times with TBST
and developed with 100 .mu.l of Fast-OPD substrate (Sigma #P9187).
Colour was allowed to develop for 5-10 mins after which time the
ELISA was stopped with 25 .mu.l 3M H.sub.2SO.sub.4. The absorbance
at 490 nM was read plate and antibody concentration determined by
reference to a standard curve.
Example 5
Antibody Competition ELISA Protocol
[0160] GST-human NOGO-A56 (SEQ ID: 76) at 0.1-1.0 .mu.g/ml in PBS
was coated onto Nunc Immunosorp plates (100 .mu.l per well) at
4.degree. C. overnight or at 37.degree. C. for 1 hour. Wells were
rinsed three times with PBS then incubated with 1% BSA in PBS
(block buffer) to block non-specific binding sites at room
temperature for 2 hours. In parallel, a 50:50 mix of antibodies was
made. Murine antibody 2A10 was added to a final concentration of
either 0.5 or 1.0 mcg/ml in block buffer. Chimeric antibodies
(mouse variable regions cloned onto a human IgG1 Fc mutated
constant region) were added to a final concentration of 0-25 mcg/ml
in block buffer. The block buffer was removed from the plates and
100 .mu.l of the 50:50 mix of antibodies was added for 1 hour at
room temperature. Wells were washed three times with PBS then
incubated with 100 .mu.l of rabbit polyclonal anti-mouse
immunoglobulins peroxidase conjugate (diluted 1:2000 in block
buffer, DakoCytomation #P0260) for 1 hour at room temperature. The
wells were washed three times with PBS and then incubated with 100
.mu.l OPD peroxidase substrate (Sigma #P9187) or TMB substrate
(Sigma #T8665) per well for 10-30 minutes. The colour reaction was
stopped by the addition of 25 .mu.l concentrated H.sub.2SO.sub.4.
Optical density at 490 nm (OPD) or 450 nm (TMB) was measured using
a plate reader.
[0161] In the first experiment (FIG. 7A), plates were coated with
GST-human NOGO-A56 at 0.5 .mu.g/ml in PBS overnight at 4.degree. C.
and the plates were developed with TMB substrate. In this
experiment, the murine antibody 2A10 was assessed in combination
with HcLc (the chimeric form of 2A10), 11C7, an isotype control
chimeric antibody and a blank control. In the second experiment
(FIG. 7B), plates were coated with GST-human NOGO-A56 at 0.5
.mu.g/ml in PBS at 37.degree. C. for 1 hour and the plates were
developed with OPD substrate. In this experiment, the murine
antibody 2A10 was assessed in combination with HcLc, 11C7, an
isotype control and H16L18.
Example 6
Production of NOGO-A Fragment (NOGO-A56, SEQ. I.D. NO:76)
[0162] A cDNA sequence encoding amino acids 586-785
(MQESLYPAAQLCPSFEESEATPSPVLPDIVMEAPLNSAVPSAGASVIQPSSSPLEASS
VNYESIKHEPENPPPYEEAMSVSLKKVSGIKEEIKEPENINAALQETEAPYISIACDL
IKETKLSAEPAPDFSDYSEMAKVEQPVPDHSELVEDSSPDSEPVDLFSDDSIPDVPQK
QDETVMLVKESLTETSFESMIEYENKE--SEQ.I.D.NO:76) of human NOGO-A was
cloned into the BamHI-XhoI sites of pGEX-6P1 to generate a
GST-tagged fusion protein designated NOGO-A56. Plasmid was
expressed in BL21 cells in 2.times.TY medium with 100 .mu.g/ml
ampicillin following induction with IPTG to 0.5 mM at 37 C for 3
hours. Cell pellets were lysed by sonication and the fusion protein
purified using Glutathione-sepharose (Amersham Pharmacia) following
manufacturers instructions. Purified protein was eluted using
reduced glutathione and extensively dialysed against PBS,
quantitated using BSA standards and a BioRad coomassie based
protein assay and then stored in aliquots at -80 C.
Example 7
BiaCore Analysis of Humanised Anti NOGO Monoclonal Antibodies
[0163] The binding kinetics of the anti-NOGO monoclonal antibody
(mAb) to recombinantly expressed human NOGO-A (GST-human NOGO-A56)
was analysed using the Biacore3000 biosensor. The hNOGO-A chip was
prepared as follows:
Method
[0164] hNOGO (GST-human NOGO-A56) was immobilised to a CM5 chip by
primary amine coupling using the Biacore Wizard program designed
for targeted immobilisation levels. The CM5 sensor surface was
activated by passing a solution of 50 mM N-hydroxy-succinimide
(NHS) and 200 mM N-ethyl-N'-dimethylaminopropyl carbonide (EDC).
Then hNOGO in sodium acetate buffer, pH5.0 or pH 4.5, was passed
over the chip and immobilised. After immobilisation was complete
any still activated esters were blocked by an injection of 1M
ethanolamine hydrochloride, pH8.5.
[0165] The anti-NOGO mAbs were diluted down in HBS-EP (10 mM HEPES,
pH 7.4, 150 mM NaCl, 3 mM EDTA, and 0.005% P-20 surfactant) and
binding studies were carried out at range of defined antibody
concentrations. All runs were referenced against a blanked sensor
surface (one that had been activated and blocked as described
earlier but had no addition of ligand). Analysis of binding was
carried out using the BIAevaluation kinetic analysis software
version 4.1. Biacore analysis of other antibodies of the invention
essentially followed the same protocol as described herein.
TABLE-US-00008 TABLE 8 Results Mean (+/-standard deviation) of four
separate experiments Antibody ka (1/Ms) kd (1/s) KD (nM) HcLc
2.70e6 (2.7e5) 3.78e-3 (7.0e-4) 1.41 (0.3) H6L13 1.82e6 (3.5e5)
1.37e-2 (2.0e-3) 7.68 (1.2) H16L16 4.37e6 (4.5e5) 5.54e-3 (1.4e-3)
1.26 (0.2) H16L18 4.18e6 (4.1e5) 5.52e-3 (9.0e-4) 1.33 (0.2) H17L16
3.38e6 (1.3e5) 6.10e-3 (1.3e-3) 1.82 (0.4) H18L16 3.64e6 (3.5e5)
5.86e-3 (9.0e-4) 1.62 (0.3) H1L11 1.73e6 (1.7e5) 3.14e-2 (4.2e-3)
18.6 (3.7)
TABLE-US-00009 TABLE 9 Results Results shown are from a single
experiment with the exception of HcLc and H6Lc where the values
shown are the mean (+/-standard deviation) of two separate
experiments Antibody (number of independent runs) ka (1/Ms) kd
(1/s) KD (nM) HcLc (2) 3.52e6 (2.8e5) 3.46e-3 (7.9e-4) 0.995 (0.3)
HcL11 3.78e6 1.34e-2 3.55 H6Lc (2) 2.17e6 (3.8e5) 2.84e-3 (1.5e-3)
2.21 (0.3) HcL13 4.8e6 9.38e-3 1.98 H6L13 2.95e6 2.33e-2 7.9 H6L6
1.2e6 2.54e-2 21.4 H10L13 1.64e6 2.12e-2 12.95 H700L11 1.19e6
2.33e-2 19.45 H5L6 1.5e6 4.7e-2 30.3 11C7 1.44e6 8.25e-5 0.057
TABLE-US-00010 TABLE 10 Results Results shown are from a single
experiment Antibody ka (1/Ms) kd (1/s) KD (nM) H6L13 1.77e6 1.3e-2
7.3 H16L16 5.47e6 5.69e-3 1.0 H14L16 2.05e6 7.39e-3 3.6 H14L18
2.41e6 8.69e-3 3.6 H1L11 1.33e6 2.84e-2 21.3 HcLc 2.62e6 4.12e-3
1.6
Example 8
BiaCore Analysis of Humanised Anti NOGO Monoclonal Antibodies Using
Off-Rate Ranking
[0166] The hNOGO chip was prepared as for kinetic analysis. Cell
supernatants where taken directly from transient transfections of
CHO--K1 cells. These were passed directly over the sensor surface
and the interaction measured. A mock transfected cell supernatant
was used for double referencing to remove any artefacts due to the
tissue culture media. All runs were referenced against a blanked
sensor surface (one that had been activated and blocked as
described earlier but had no addition of ligand). Analysis of
binding was carried out using the BIAevaluation kinetic analysis
software version 4.1.
TABLE-US-00011 TABLE 11 Results Results shown are from a single
experiment with the exception of H6L13, H16L16, H16L18, H1L11, HcLc
and H18L16 where the values shown are the mean (+/-standard
deviation) of two or three separate experiments Antibody (number of
independent runs) kd (1/s) H14L13 1.38e-2 H15L13 9.65e-3 H16L13
9.07e-3 H17L13 9.31e-3 H18L13 9.07e-3 H6L13 (.times.3) 1.73e-2
(4.8e-3) H16L16 (.times.3) 6.64e-3 (9.2e-4) H16L18 (.times.3)
6.09e-3 (7.4e-4) H1L11 (.times.3) 4.03e-2 (1.8e-2) HcLc (.times.2)
3.76e-3 (7.1e-4) H15L16 6.04e-3 H14L16 8.9e-3 H18L16 (.times.2)
6.87e-3 (7.5e-4) H14L18 8.35e-3 H15L18 5.94e-3 H18L18 5.8e-3 H6L17
1.58e-2 H6L18 1.06e-2 H6L14 4.57e-2 H6L15 2.11e-2 H6L16 1.14e-2
Example 9
FACS Analysis of Humanised Anti NOGO Monoclonal Antibodies
[0167] IMR32 human neuroblastoma cells were re-suspended in FACS
staining buffer (PBS+4% heat inactivated FCS) at a density of
10.sup.6 cells per ml. 100 .mu.l of this suspension was transferred
to wells of a 96 well round bottomed microplate. 100 .mu.l of "Fix
& Perm" Medium A (Caltag Laboratories, GAS001 S-100) was added
to each well and the plate incubated at room temperature for 15
mins. The cells were pelleted was washed twice in FACS staining
buffer. Following washing, the cells were re-suspended in 50 .mu.l
of a solution of the anti-NOGO antibodies or an isotype matched
control antibody at a concentration of 2.times. the final
concentration (0-200 .mu.g/ml in FACS staining buffer). 50 .mu.l of
"Fix & Perm" Medium B (Caltag Laboratories GAS002S-100) was
added and the plate incubated on ice for 1 hour. Cells were washed
twice in FACS staining buffer before being re-suspended in 100
.mu.l of a solution of a PE conjugated anti human .gamma.1 specific
goat F(ab').sub.2 (Sigma P-8047) at a dilution of 1/50. Cells
incubated for 1 hour on ice. The cells were pelleted and washed 3
times in FACS staining buffer and cells re-suspended in 100 .mu.l
the same buffer. 100 .mu.l of "Fix & Perm" Medium B was added
to fix the cells. The degree of staining was determined by flow
cytometry using a Becton Dickinson FACScan flow cytometer. The
isotype matched controls were used as reference.
[0168] Results are shown in FIG. 6 A to F. The totality of the data
shown illustrates that the HcLc antibody (of 2A10) gives a strong
signal in this FACS assay, which indicates strong binding to human
cell expressed NOGO. The data also shows that humanised versions of
this chimera can retain this property. The 2A10 chimeric antibody,
and the best humanised versions, thereof consistently outperform
11C7 in this assay.
TABLE-US-00012 NOGO antibody sequences Summary (Table 12) Sequence
identifier (SEQ.I.D.NO) amino acid Polynucleotide Description
sequence sequence 2A10, CDR-H1 1 -- 2A10, CDR-H2 2 -- 2A10, CDR-H3
3 -- 2A10, CDR-L1 4 -- 2A10, CDR-L2 5 -- 2A10, CDR-L3 6 -- 2A10, VH
(murine) 7 41 2A10, VL (murine) 8 42 Chimeric heavy chain Hc 9 43
Chimeric light chain Lc 10 44 2A10 VH humanised construct H5 11 45
2A10 VH humanised construct H6 12 46 2A10 VH humanised construct
H700 13 47 2A10 VH humanised construct H14 14 48 2A10 VH humanised
construct H15 15 49 2A10 VH humanised construct H16 16 50 2A10 VH
humanised construct H17 17 51 2A10 VH humanised construct H18 18 52
2A10 VL humanised construct L6 19 53 2A10 VL humanised construct
L13 20 54 2A10 VL humanised construct L14 21 55 2A10 VL humanised
construct L15 22 56 2A10 VL humanised construct L16 23 57 2A10 VL
humanised construct L17 24 58 2A10 VL humanised construct L18 25 59
2A10 heavy chain humanised construct H5 26 60 2A10 heavy chain
humanised construct H6 27 61 2A10 heavy chain humanised construct
28 62 H700 2A10 heavy chain humanised construct 29 63 H14 2A10
heavy chain humanised construct 30 64 H15 2A10 heavy chain
humanised construct 31 65 H16 2A10 heavy chain humanised construct
32 66 H17 2A10 heavy chain humanised construct 33 67 H18 2A10 light
chain humanised construct L6 34 68 2A10 light chain humanised
construct L13 35 69 2A10 light chain humanised construct L14 36 70
2A10 light chain humanised construct L15 37 71 2A10 light chain
humanised construct L16 38 72 2A10 light chain humanised construct
L17 39 73 2A10 light chain humanised construct L18 40 74 Campath
leader sequence 75 -- Amino acids 586-785 of human NOGO A 76 --
(NOGO-A56) 2A10 VH humanised construct H1 77 81 2A10 VL humanised
construct L11 78 82 2A10 heavy chain humanised construct H1 79 83
2A10 light chain humanised construct L11 80 84 2A10 VH humanised
construct H19 85 99 2A10 VH humanised construct H20 86 100 2A10 VH
humanised construct H21 87 101 2A10 VH humanised construct H22 88
102 2A10 VH humanised construct H23 89 103 2A10 VH humanised
construct H24 90 104 2A10 VH humanised construct H25 91 105 2A10
heavy chain humanised construct 92 106 H19 2A10 heavy chain
humanised construct 93 107 H20 2A10 heavy chain humanised construct
94 108 H21 2A10 heavy chain humanised construct 95 109 H22 2A10
heavy chain humanised construct 96 110 H23 2A10 heavy chain
humanised construct 97 111 H24 2A10 heavy chain humanised construct
98 112 H25
TABLE-US-00013 SEQ ID 1: 2A10 CDR-H1 SYWMH SEQ ID 2: 2A10 CDR-H2
NINPSNGGTNYNEKFKS SEQ ID 3: 2A10 CDR-H3 GQGY SEQ ID 4: 2A10 CDR-L1
RSSKSLLYKDGKTYLN SEQ ID 5: 2A10 CDR-L2 LMSTRAS SEQ ID 6: 2A10
CDR-L3 QQLVEYPLT SEQ ID 7: 2A10, VH (murine)
QVQLQQPGTELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCELGQ GYWGQGTTLTVSS
SEQ ID 8: 2A10, VL (murine)
DIVITQDELSNPVTSGESVSISCRSSKSLLYKDGKTYLNWFLQRPGQSPQ
LLIYLMSTRASGVSDRFSGSGSGTDFTLEISRVKAEDVGVYYCQQLVEYP LTFGAGTKLELK SEQ
ID 9: Chimeric heavy chain Hc
MGWSCIILFLVAAATGVHSQVQLQQPGTELVKPGASVKLSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATLTVDKSSSTAYM
QLSSLTSEDSAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 10: Chimeric light chain Lc
MRCSLQFLGVLMFWISGVSGDIVITQDELSNPVTSGESVSISCRSSKSLL
YKDGKTYLNWFLQRPGQSPQLLIYLMSTRASGVSDRFSGSGSGTDFTLEI
SRVKAEDVGVYYCQQLVEYPLTFGAGTKLELKRTVAAPSVFIFPPSDEQL
KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL
SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 11: 2A10 VH
humanised construct H5
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQGLEWMGN
INPSNGGTNYNEKFKSRVTMTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 12: 2A10 VH humanised construct H6
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQGLEWIGN
INPSNGGTNYNEKFKSRATMTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 13: 2A10 VH humanised construct H700
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQGLEWIGN
INPSNGGTNYNEKFKSRATLTVDKSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 14: 2A10 VH humanised construct H14
QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQAPGQGLEWIGN
INPSNGGTNYNEKFKSRATLTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 15: 2A10 VH humanised construct H15
QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATLTVDTSSSTAYMELSSLRSEDSAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 16: 2A10 VH humanised construct H16
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATLTVDKSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 17: 2A10 VH humanised construct H17
QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATLTVDKSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 18: 2A10 VH humanised construct H18
QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATLTVDKSSSTAYMELSSLRSEDSAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 19: 2A10 VL humanised construct L6
DIVMTQSPLSLPVTPGEPASISCRSSKSLLYKDGKTYLNWFLQRPGQSPQ
LLIYLMSTRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGGGTKVEIK SEQ
ID 20: 2A10 VL humanised construct L13
DIVMTQSPLSLPVTLGQPASISCRSSKSLLYKDGKTYLNWFQQRPGQSPQ
LLIYLMSTRASGVPDRFSGGGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 21: 2A10 VL humanised construct L14
DIVMTQSPLSLPVTLGQPASISCRSSKSLLYKDGKTYLNWFLQRPGQSPQ
LLIYLMSTRASGVPDRFSGGGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 22: 2A10 VL humanised construct L15
DIVMTQSPLSLPVTLGQPASISCRSSKSLLYKDGKTYLNWFLQRPGQSPQ
LLIYLMSTRASGVSDRFSGGGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 23: 2A10 VL humanised construct L16
DIVMTQSPLSNPVTLGQPVSISCRSSKSLLYKDGKTYLNWFLQRPGQSPQ
LLIYLMSTRASGVPDRFSGGGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 24: 2A10 VL humanised construct L17
DIVMTQDPLSLPVTLGQPASISCRSSKSLLYKDGKTYLNWFQQRPGQSPQ
LLIYLMSTRASGVSDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 25: 2A10 VL humanised construct L18
DIVMTQDPLSNPVTLGQPVSISCRSSKSLLYKDGKTYLNWFLQRPGQSPQ
LLIYLMSTRASGVSDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 26: 2A10 heavy chain humanised construct H5
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQAPGQGLEWMGNINPSNGGTNYNEKFKSRVTMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 27: 2A10 heavy chain humanised construct H6
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQAPGQGLEWIGNINPSNGGTNYNEKFKSRATMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 28: 2A10 heavy chain humanised construct H700
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQAPGQGLEWIGNINPSNGGTNYNEKFKSRATLTVDKSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 29: 2A10 heavy chain humanised construct H14
MGWSCIILFLVATATGVHSQVQLVQSGAEVVKPGASVKLSCKASGYTFTS
YWMHWVKQAPGQGLEWIGNINPSNGGTNYNEKFKSRATLTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 30: 2A10 heavy chain humanised construct H15
MGWSCIILFLVATATGVHSQVQLVQSGAEVVKPGASVKLSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATLTVDTSSSTAYM
ELSSLRSEDSAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 31: 2A10 heavy chain humanised construct H16
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATLTVDKSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 32: 2A10 heavy chain humanised construct H17
MGWSCIILFLVATATGVHSQVQLVQSGAEVVKPGASVKLSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATLTVDKSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 33: 2A10 heavy chain humanised construct H18
MGWSCIILFLVATATGVHSQVQLVQSGAEVVKPGASVKLSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATLTVDKSSSTAYM
ELSSLRSEDSAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 34: 2A10 light chain humanised construct L6
MGWSCIILFLVATATGVHSDIVMTQSPLSLPVTPGEPASISCRSSKSLLY
KDGKTYLNWFLQRPGQSPQLLIYLMSTRASGVPDRFSGSGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 35: 2A10 light chain
humanised construct L13
MGWSCIILFLVATATGVHSDIVMTQSPLSLPVTLGQPASISCRSSKSLLY
KDGKTYLNWFQQRPGQSPQLLIYLMSTRASGVPDRFSGGGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 36: 2A10 light chain
humanised construct L14
MGWSCIILFLVATATGVHSDIVMTQSPLSLPVTLGQPASISCRSSKSLLY
KDGKTYLNWFLQRPGQSPQLLIYLMSTRASGVPDRFSGGGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 37: 2A10 light chain
humanised construct L15
MGWSCIILFLVATATGVHSDIVMTQSPLSLPVTLGQPASISCRSSKSLLY
KDGKTYLNWFLQRPGQSPQLLIYLMSTRASGVSDRFSGGGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 38: 2A10 light chain
humanised construct L16
MGWSCIILFLVATATGVHSDIVMTQSPLSNPVTLGQPVSISCRSSKSLLY
KDGKTYLNWFLQRPGQSPQLLIYLMSTRASGVPDRFSGGGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 39: 2A10 light chain
humanised construct L17
MGWSCIILFLVATATGVHSDIVMTQDPLSLPVTLGQPASISCRSSKSLLY
KDGKTYLNWFQQRPGQSPQLLIYLMSTRASGVSDRFSGSGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 40: 2A10 light chain
humanised construct L18
MGWSCIILFLVATATGVHSDIVMTQDPLSNPVTLGQPVSISCRSSKSLLY
KDGKTYLNWFLQRPGQSPQLLIYLMSTRASGVSDRFSGSGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 41: PN encoding 2A10,
VH (murine) SEQ ID: 7
CAGGTCCAACTGCAGCAGCCTGGGACTGAACTGGTGAAGCCTGGGGCTTC
AGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGA
TGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAA
GGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCA
GCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGAACTGGGACAG
GGCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA SEQ ID 42: PN encoding
2A10, VL (murine) SEQ ID: 8
GATATTGTGATAACCCAGGATGAACTCTCCAATCCTGTCACTTCTGGAGA
ATCAGTTTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCTGCAGAGACCAGGACAATCTCCTCAG
CTCCTGATCTATTTGATGTCCACCCGTGCATCAGGAGTCTCAGACCGGTT
TAGTGGCAGTGGGTCAGGAACAGATTTCACCCTGGAAATCAGTAGAGTGA
AGGCTGAGGATGTGGGTGTGTATTACTGTCAACAACTTGTAGAGTATCCG
CTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA SEQ ID 43: PN encoding
Chimeric heavy chain Hc SEQ ID: 9
ATGGGATGGAGCTGTATCATCCTCTTTTTGGTAGCAGCAGCTACAGGTGT
CCACTCCCAGGTCCAACTGCAGCAGCCTGGGACTGAACTGGTGAAGCCTG
GGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGC
TACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGAT
TGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCA
AGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATG
CAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGAACT
GGGACAGGGCTACTGGGGCCAAGGCACACTAGTCACAGTCTCCTCAGCCT
CCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACC
TCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA
ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA
CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTG
GTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGT
GAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAAT
CTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCGCG
GGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCAT
GATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT
AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT
GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGT
ACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACC
ATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCC
CCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGG
TCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG
CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG
CTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGC
AGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC
TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID 44: PN encoding
Chimeric light chain Lc SEQ ID: 10
ATGAGGTGCTCTCTTCAGTTTCTGGGGGTGCTTATGTTCTGGATCTCTGG
AGTCAGTGGGGATATTGTGATAACCCAGGATGAACTCTCCAATCCTGTCA
CTTCTGGAGAATCAGTTTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTA
TATAAGGATGGGAAGACATACTTGAATTGGTTTCTGCAGAGACCAGGACA
ATCTCCTCAGCTCCTGATCTATTTGATGTCCACCCGTGCATCAGGAGTCT
CAGACCGGTTTAGTGGCAGTGGGTCAGGAACAGATTTCACCCTGGAAATC
AGTAGAGTGAAGGCTGAGGATGTGGGTGTGTATTACTGTCAACAACTTGT
AGAGTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGTA
CGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTG
AAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAG
AGAGGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACT
CCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC
AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTA
CGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCT
TCAACAGGGGAGAGTGTTAG SEQ ID 45: PN encoding 2A10 VH humanised
construct H5 SEQ ID: 11
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTC
AGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAG
AGTCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 46: PN encoding 2A10
VH humanised construct H6 SEQ ID: 12
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTC
AGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAG
AGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 47: PN encoding 2A10
VH humanised construct H700 SEQ ID: 13
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTC
AGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAG
AGCCACCCTGACCGTAGACAAATCCACGAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 48: PN encoding 2A10
VH humanised construct H14 SEQ ID: 14
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCTGGGGCCTC
AGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGAAACAGGCCCCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAG
AGCCACCCTCACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 49: PN encoding 2A10
VH humanised construct H15 SEQ ID: 15
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCTGGGGCCTC
AGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAA
AGCCACCCTCACCGTCGACACGTCCAGCAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACAGCGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 50: PN encoding 2A10
VH humanised construct H16 SEQ ID: 16
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTC
AGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAA
AGCCACCCTCACCGTCGACAAATCCACGAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 51: PN encoding 2A10
VH humanised construct H17 SEQ ID: 17
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCTGGGGCCTC
AGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAA
AGCCACCCTCACCGTCGACAAATCCACGAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 52: PN encoding 2A10
VH humanised construct H18 SEQ ID: 18
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCTGGGGCCTC
AGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGGA
TGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGGATCGGAAAT
ATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGCAA
AGCCACCCTCACCGTCGACAAATCCAGCAGCACAGCCTACATGGAGCTGA
GCAGCCTGAGATCTGAGGACAGCGCCGTGTATTACTGTGAACTGGGACAG
GGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 53: PN encoding 2A10
VL humanised construct L6 SEQ ID: 19
GATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGA
GCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTCCTGCAGCGACCAGGGCAGTCTCCACAG
CTCCTGATCTATTTGATGTCCACCCGTGCATCAGGGGTCCCTGACAGGTT
CAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGCAGAGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTCGGCGGAGGGACCAAGGTGGAGATCAAA SEQ ID 54: PN encoding 2A10 VL
humanised construct L13 SEQ ID: 20
GATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACA
GCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAGACAGATT
CAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 55: PN encoding 2A10 VL
humanised construct L14 SEQ ID: 21
GATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACA
GCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAGACAGATT
CAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 56: PN encoding 2A10 VL
humanised construct L15 SEQ ID: 22
GATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACA
GCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCAGCGACAGATT
CAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 57: PN encoding 2A10 VL
humanised construct L16 SEQ ID: 23
GATATTGTGATGACCCAGTCTCCACTCTCCAACCCCGTCACCCTTGGACA
GCCGGTCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAGACAGATT
AGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGGC
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 58: PN encoding 2A10 VL
humanised construct L17 SEQ ID: 24
GATATTGTGATGACCCAGGACCCACTCTCCCTGCCCGTCACCCTTGGACA
GCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCAGCGACAGATT
CAGCGGCAGCGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 59: PN encoding 2A10 VL
humanised construct L18 SEQ ID: 25
GATATTGTGATGACCCAGGACCCACTCTCCAACCCCGTCACCCTTGGACA
GCCGGTCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCAGCGACAGATT
CAGCGGCAGCGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 60: PN encoding 2A10
heavy chain humanised construct H5 SEQ ID: 26
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTG
GGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGC
TACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGAT
GGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCA
AGAGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATG
GAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACT
GGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAGCCT
CCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACC
TCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA
ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA
CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTG
GTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGT
GAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAAT
CTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCGCG
GGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCAT
GATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT
AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT
GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGT
ACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACC
ATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCC
CCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGG
TCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG
CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG
CTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGC
AGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC
TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID 61: PN encoding 2A10
heavy chain humanised construct H6 SEQ ID: 27
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTG
GGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGC
TACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGAT
CGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCA
AGAGCAGAGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATG
GAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACT
GGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAGCCT
CCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACC
TCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA
ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA
CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTG
GTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGT
GAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAAT
CTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCGCG
GGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCAT
GATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT
AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT
GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGT
ACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACC
ATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCC
CCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGG
TCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG
CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG
CTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGC
AGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC
TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID 62: PN encoding 2A10
heavy chain humanised construct H700 SEQ ID: 28
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTG
GGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGC
TACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGAT
CGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCA
AGAGCAGAGCCACCCTGACCGTAGACAAATCCACGAGCACAGCCTACATG
GAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACT
GGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAGCCT
CCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACC
TCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA
ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA
CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTG
GTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGT
GAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAAT
CTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCGCG
GGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCAT
GATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT
AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT
GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGT
ACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACC
ATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCC
CCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGG
TCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG
CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG
CTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGC
AGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC
TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID 63: PN encoding 2A10
heavy chain humanised construct H14 SEQ ID: 29
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCT
GGGGCCTCAGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAACAGGCCCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAGAGCCACCCTCACCAGGGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
64: PN encoding 2A10 heavy chain humanised construct H15 SEQ ID: 30
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCT
GGGGCCTCAGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAAGCCACCCTCACCGTCGACACGTCCAGCAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACAGCGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
65: PN encoding 2A10 heavy chain humanised construct H16 SEQ ID: 31
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAAGCCACCCTCACCGTCGACAAATCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
66: PN encoding 2A10 heavy chain humanised construct H17 SEQ ID: 32
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCT
GGGGCCTCAGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAAGCCACCCTCACCGTCGACAAATCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
67: PN encoding 2A10 heavy chain humanised construct H18 SEQ ID: 33
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGGTCAAGCCT
GGGGCCTCAGTGAAGCTCTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAACAGCGACCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAAGCCACCCTCACCGTCGACAAATCCAGCAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACAGCGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
68: PN encoding 2A10 light chain humanised construct L6 SEQ ID: 34
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCC
CTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTCCTGCAGCGACCAGGGCAGTC
TCCACAGCTCCTGATCTATTTGATGTCCACCCGTGCATCAGGGGTCCCTG
ACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGC
AGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACG
GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA
ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAG
AGGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCC
CAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA
GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC
GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCT
TCAACAGGGGAGAGTGTTAG SEQ ID 69: PN encoding 2A10 light chain
humanised construct L13 SEQ ID: 35
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCC
TTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAG
ACAGATTCAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACGG
TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA
GGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCCC
AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC
AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC
CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTC
AACAGGGGAGAGTGTTAG SEQ ID 70: PN encoding 2A10 light chain
humanised construct L14 SEQ ID: 36
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCC
TTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAG
ACAGATTCAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACGG
TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA
GGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCCC
AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC
AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC
CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTC
AACAGGGGAGAGTGTTAG SEQ ID 71: PN encoding 2A10 light chain
humanised construct L15 SEQ ID: 37
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCC
TTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCAGCG
ACAGATTCAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACG
GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA
ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAG
AGGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCC
CAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA
GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC
GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCT
TCAACAGGGGAGAGTGTTAG SEQ ID 72: PN encoding 2A10 light chain
humanised construct L16 SEQ ID: 38
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACCCAGTCTCCACTCTCCAACCCCGTCACCC
TTGGACAGCCGGTCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAG
ACAGATTCAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACGG
TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA
GGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCCC
AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC
AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC
CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTC
AACAGGGGAGAGTGTTAG SEQ ID 73: PN encoding 2A10 light chain
humanised construct L17 SEQ ID: 39
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACCCAGGACCCACTCTCCCTGCCCGTCACCC
TTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCAGCG
ACAGATTCAGCGGCAGCGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACG
GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA
ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAG
AGGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCC
CAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA
GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC
GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCT
TCAACAGGGGAGAGTGTTAG SEQ ID 74: PN encoding 2A10 light chain
humanised construct L18 SEQ ID: 40
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATGACCCAGGACCCACTCTCCAACCCCGTCACCC
TTGGACAGCCGGTCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCTCCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCAGCG
ACAGATTCAGCGGCAGCGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACGG
TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA
GGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCCC
AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC
AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC
CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTC
AACAGGGGAGAGTGTTAG SEQ ID 75: Campath leader sequence
MGWSCIILFLVATATGVHS SEQ ID 76: Amino acids 586-785 of human NOGO A
(NOGO-A56) MQESLYPAAQLCPSFEESEATPSPVLPDIVMEAPLNSAVPSAGASVIQPS
SSPLEASSVNYESIKHEPENPPPYEEAMSVSLKKVSGIKEEIKEPENINA
ALQETEAPYISIACDLIKETKLSAEPAPDFSDYSEMAKVEQPVPDHSELV
EDSSPDSEPVDLFSDDSIPDVPQKQDETVMLVKESLTETSFESMIEYEN KE SEQ ID 77:
2A10 VH humanised construct H1
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQGLEWMGN
INPSNGGTNYNEKFKSRVTMTRDTSTSTVYMELSSLRSEDTAVYYCELGQ
GYWGQGTLVTVSS SEQ ID 78: 2A10 VL humanised construct L11
DIVITQSPLSLPVTLGQPASISCRSSKSLLYKDGKTYLNWFQQRPGQSPQ
LLIYLMSTRASGVPDRFSGGGSGTDFTLKISRVEAEDVGVYYCQQLVEYP LTFGQGTKLEIK SEQ
ID 79: 2A10 heavy chain humanised construct H1
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQAPGQGLEWMGNINPSNGGTNYNEKFKSRVTMTRDTSTSTVYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 80: 2A10 light chain humanised construct L11
MGWSCIILFLVATATGVHSDIVITQSPLSLPVTLGQPASISCRSSKSLLY
KDGKTYLNWFQQRPGQSPQLLIYLMSTRASGVPDRFSGGGSGTDFTLKIS
RVEAEDVGVYYCQQLVEYPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS
STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID 81: PN encoding 2A10
VH humanised construct H1 SEQ ID: 77
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 82: PN encoding
2A10 VL humanised construct L11 SEQ ID: 78
GATATTGTGATAACCCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACA
GCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATG
GGAAGACATACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCACAG
CTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAGACAGATT
CAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGG
AGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGAGTATCCG
CTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID 83: PN encoding 2A10
humanised heavy chain H1 SEQ ID: 79
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGG
ATGGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
84: PN encoding 2A10 humanised light chain construct L11 SEQ ID: 90
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCGATATTGTGATAACCCAGTCTCCACTCTCCCTGCCCGTCACCC
TTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATAT
AAGGATGGGAAGACATACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATC
TCCACAGCTCCTAATTTATTTGATGTCCACCCGTGCATCTGGGGTCCCAG
ACAGATTCAGCGGCGGTGGGTCAGGCACTGATTTCACACTGAAAATCAGC
AGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCCAACAACTTGTAGA
GTATCCGCTCACGTTTGGCCAGGGGACCAAGCTGGAGATCAAACGTACGG
TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA
GGCCAAAGTACAGTGGAAGGTGGACAACGCCCTCCAATCGGGTAACTCCC
AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC
AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC
CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTC
AACAGGGGAGAGTGTTAG SEQ ID 85: 2A10 VH humanised construct H19
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQRPGQGLEWIGN
INPSNGGTNYNEKFKSRATMTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 86: 2A10 VH humanised construct H20
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQGLEWIGN
INPSNGGTNYNEKFKSKATMTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 87: 2A10 VH humanised construct H21
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATMTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 88: 2A10 VH humanised construct H22
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATMTRDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 89: 2A10 VH humanised construct H23
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATMTVDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 90: 2A10 VH humanised construct H24
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVKQRPGQGLEWIGN
INPSNGGTNYNEKFKSKATLTVDTSTSTAYMELSSLRSEDTAVYYCELGQ GYWGQGTLVTVSS
SEQ ID 91: 2A10 VH humanised construct H25
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQRPGQGLEWMG
NINPSNGGTNYNEKFKSRVTMTRDTSTSTAYMELSSLRSEDTAVYYCELG QGYWGQGTLVTVSS
SEQ ID 92: 2A10 heavy chain humanised construct H19
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQRPGQGLEWIGNINPSNGGTNYNEKFKSRATMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 93: 2A10 heavy chain humanised construct H20
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQAPGQGLEWIGNINPSNGGTNYNEKFKSKATMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 94: 2A10 heavy chain humanised construct H21
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQRPGQGLEWIGNINPSNGGTNYNEKFKSKATMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 95: 2A10 heavy chain humanised construct H22
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 96: 2A10 heavy chain humanised construct H23
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATMTVDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 97: 2A10 heavy chain humanised construct H24
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVKQRPGQGLEWIGNINPSNGGTNYNEKFKSKATLTVDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 98: 2A10 heavy chain humanised construct H25
MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTS
YWMHWVRQRPGQGLEWMGNINPSNGGTNYNEKFKSRVTMTRDTSTSTAYM
ELSSLRSEDTAVYYCELGQGYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELA
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQ
ID 99: PN encoding 2A10 VH humanised construct H19 SEQ ID: 85
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGCGACAGAGGCCTGGACAAGGGCTTGAGTGGATCGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AGAGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 100: PN encoding
2A10 VH humanised construct H20 SEQ ID: 86
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATCGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AAGGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 101: PN encoding
2A10 VH humanised construct H21 SEQ ID: 87
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGCGACAGAGGCCTGGACAAGGGCTTGAGTGGATCGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AAGGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 102: PN encoding
2A10 VH humanised construct H22 SEQ ID: 88
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGAAGCAGAGGCCTGGACAAGGGCTTGAGTGGATCGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AAGGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 103: PN encoding
2A10 VH humanised construct H23 SEQ ID: 89
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGAAGCAGAGGCCTGGACAAGGGCTTGAGTGGATCGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AAGGCCACCATGACCGTAGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 104: PN encoding
2A10 VH humanised construct H24 SEQ ID: 90
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGAAGCAGAGGCCTGGACAAGGGCTTGAGTGGATCGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AAGGCCACCCTGACCGTAGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 105: PN encoding
2A10 VH humanised construct H25 SEQ ID: 91
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCT
CAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTGG
ATGCACTGGGTGCGACAGAGGCCTGGACAAGGGCTTGAGTGGATGGGAA
ATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTTCAAGAGC
AGAGTCACCATGACCAGGGACACGTCCACGAGCACAGCCTACATGGAGCT
GAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAACTGGGA
CAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCA SEQ ID 106: PN encoding
2A10 heavy chain humanised construct H19 SEQ ID: 92
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGCGACAGAGGCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAGAGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
107: PN encoding 2A10 heavy chain humanised construct H20 SEQ ID:
93 ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAGGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
108: PN encoding 2A10 heavy chain humanised construct H21 SEQ ID:
94 ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGCGACAGAGGCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAGGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
109: PN encoding 2A10 heavy chain humanised construct H22 SEQ ID:
95 ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAGGCCACCATGACCAGGGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
110: PN encoding 2A10 heavy chain humanised construct H23 SEQ ID:
96 ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAGGCCACCATGACCGTAGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
111: PN encoding 2A10 heavy chain humanised construct H24 SEQ ID:
97 ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGGCTTGAGTGG
ATCGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAAGGCCACCCTGACCGTAGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
112: PN encoding 2A10 heavy chain humanised construct H25 SEQ ID:
98 ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGT
CCACTCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCT
GGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAG
CTACTGGATGCACTGGGTGCGACAGAGGCCTGGACAAGGGCTTGAGTGG
ATGGGAAATATTAATCCTAGCAATGGTGGTACTAACTACAATGAGAAGTT
CAAGAGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGCCTACA
TGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGAA
CTGGGACAGGGCTACTGGGGCCAGGGAACACTAGTCACAGTCTCCTCAG
CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAG
CACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTC
CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGC
GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAG
CAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA
GCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTG
AACTCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACG
TGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGC
ACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA
ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCC
CATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCA
GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC
GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA
CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT
GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAATGA SEQ ID
NO. 113 - Amino acid sequence of Marmoset NOGO-A fragment
VQDSLCPVAQLCPSFEESEATPSPVLPDIVMEAPLNSAVPSAGASAVQP
SSSPLEASSVNFESVKHEPENPPPYEEAMNVSRKKVSGIKEEIKEPESI
NAAVQETEAPYISIACDLIKETKLSAEPTPDFSSYSEMAKVEQPLPDHS
ELVEDSSPDSEPVDLFSDDSIPDVPQKQDEAVILVKETLTETSFESMIE HENK
Sequence CWU 1
1
11315PRTHomo sapien 1Ser Tyr Trp Met His1 5217PRTHomo sapien 2Asn
Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys1 5 10
15Ser34PRTHomo sapien 3Gly Gln Gly Tyr1416PRTHomo sapien 4Arg Ser
Ser Lys Ser Leu Leu Tyr Lys Asp Gly Lys Thr Tyr Leu Asn1 5 10
1557PRTHomo sapien 5Leu Met Ser Thr Arg Ala Ser1 569PRTHomo sapien
6Gln Gln Leu Val Glu Tyr Pro Leu Thr1 57113PRTMurine 7Gln Val Gln
Leu Gln Gln Pro Gly Thr Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val
Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp
Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40
45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala
Tyr65 70 75 80Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val
Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Thr
Leu Thr Val Ser 100 105 110Ser8112PRTMurine 8Asp Ile Val Ile Thr
Gln Asp Glu Leu Ser Asn Pro Val Thr Ser Gly1 5 10 15Glu Ser Val Ser
Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Lys 20 25 30Asp Gly Lys
Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser 35 40 45Pro Gln
Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser Gly Val Ser 50 55 60Asp
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Glu Ile65 70 75
80Ser Arg Val Lys Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Leu
85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
Lys 100 105 1109462PRTHomo sapien 9Met Gly Trp Ser Cys Ile Ile Leu
Phe Leu Val Ala Ala Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu
Gln Gln Pro Gly Thr Glu Leu Val Lys 20 25 30Pro Gly Ala Ser Val Lys
Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met
His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly
Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys
Phe Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser 85 90 95Thr
Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105
110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230
235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val
Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345
350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 46010239PRTHomo
sapien 10Met Arg Cys Ser Leu Gln Phe Leu Gly Val Leu Met Phe Trp
Ile Ser1 5 10 15Gly Val Ser Gly Asp Ile Val Ile Thr Gln Asp Glu Leu
Ser Asn Pro 20 25 30Val Thr Ser Gly Glu Ser Val Ser Ile Ser Cys Arg
Ser Ser Lys Ser 35 40 45Leu Leu Tyr Lys Asp Gly Lys Thr Tyr Leu Asn
Trp Phe Leu Gln Arg 50 55 60Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr
Leu Met Ser Thr Arg Ala65 70 75 80Ser Gly Val Ser Asp Arg Phe Ser
Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95Thr Leu Glu Ile Ser Arg Val
Lys Ala Glu Asp Val Gly Val Tyr Tyr 100 105 110Cys Gln Gln Leu Val
Glu Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys 115 120 125Leu Glu Leu
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro 130 135 140Pro
Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu145 150
155 160Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
Asp 165 170 175Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr
Glu Gln Asp 180 185 190Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
Leu Thr Leu Ser Lys 195 200 205Ala Asp Tyr Glu Lys His Lys Val Tyr
Ala Cys Glu Val Thr His Gln 210 215 220Gly Leu Ser Ser Pro Val Thr
Lys Ser Phe Asn Arg Gly Glu Cys225 230 23511113PRTArtificial
Sequence2A10 VH humanised construct H5 11Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met His Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser
Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
Ser 100 105 110Ser12113PRTArtificial Sequence2A10 VH humanised
construct H6 12Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Ser Tyr 20 25 30Trp Met His Trp Val Arg Gln Ala Pro Gly Gln
Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr
Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser Arg Ala Thr Met Thr Arg Asp
Thr Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr
Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105
110Ser13113PRTArtificial Sequence2A10 VH humanised construct H700
13Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser
Tyr 20 25 30Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn
Glu Lys Phe 50 55 60Lys Ser Arg Ala Thr Leu Thr Val Asp Lys Ser Thr
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln
Gly Thr Leu Val Thr Val Ser 100 105 110Ser14113PRTArtificial
Sequence2A10 VH humanised construct H14 14Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met His Trp
Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser
Arg Ala Thr Leu Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
Ser 100 105 110Ser15113PRTArtificial Sequence2A10 VH humanised
construct H14 15Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys
Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Ser Tyr 20 25 30Trp Met His Trp Val Lys Gln Arg Pro Gly Gln
Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr
Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Leu Thr Val Asp
Thr Ser Ser Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr
Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105
110Ser16113PRTArtificial Sequence2A10 VH humanised construct H16
16Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser
Tyr 20 25 30Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn
Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Thr
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln
Gly Thr Leu Val Thr Val Ser 100 105 110Ser17113PRTArtificial
Sequence2A10 VH humanised construct H17 17Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met His Trp
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser
Lys Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
Ser 100 105 110Ser18113PRTArtificial Sequence2A10 VH humanised
construct H18 18Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys
Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Ser Tyr 20 25 30Trp Met His Trp Val Lys Gln Arg Pro Gly Gln
Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr
Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Leu Thr Val Asp
Lys Ser Ser Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr
Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105
110Ser19112PRTArtificial Sequence2A10 VL humanised construct L6
19Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly1
5 10 15Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr
Lys 20 25 30Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly
Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser
Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val
Tyr Tyr Cys Gln Gln Leu 85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Gly
Gly Thr Lys Val Glu Ile Lys 100 105 11020112PRTArtificial
Sequence2A10 VL humanised construct L13 20Asp Ile Val Met Thr Gln
Ser Pro Leu Ser Leu Pro Val Thr Leu Gly1 5 10 15Gln Pro Ala Ser Ile
Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Lys 20 25 30Asp Gly Lys Thr
Tyr Leu Asn Trp Phe Gln Gln Arg Pro Gly Gln Ser 35 40 45Pro Gln Leu
Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60Asp Arg
Phe Ser Gly Gly Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75
80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Leu
85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys 100 105 11021112PRTArtificial Sequence2A10 VL humanised
construct L14 21Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val
Thr Leu Gly1 5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser
Leu Leu Tyr Lys 20 25 30Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln
Arg Pro Gly Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr
Arg Ala Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Gly Gly Ser Gly
Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp
Val Gly Val Tyr Tyr Cys Gln Gln Leu 85 90 95Val Glu Tyr Pro Leu Thr
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
11022112PRTArtificial Sequence2A10 VL humanised construct L15 22Asp
Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly1 5 10
15Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Lys
20 25 30Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln
Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser Gly
Val Ser 50 55 60Asp Arg Phe Ser Gly Gly Gly Ser Gly Thr Asp Phe Thr
Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
Tyr Cys Gln Gln Leu 85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly
Thr Lys Leu Glu Ile Lys 100 105 11023112PRTArtificial Sequence2A10
VL humanised construct L16 23Asp Ile Val Met Thr Gln Ser Pro Leu
Ser Asn Pro Val Thr Leu Gly1 5 10 15Gln Pro
Val Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Lys 20 25 30Asp
Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser 35 40
45Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser Gly Val Pro
50 55 60Asp Arg Phe Ser Gly Gly Gly Ser Gly Thr Asp Phe Thr Leu Lys
Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys
Gln Gln Leu 85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys
Leu Glu Ile Lys 100 105 11024112PRTArtificial Sequence2A10 VL
humanised construct L17 24Asp Ile Val Met Thr Gln Asp Pro Leu Ser
Leu Pro Val Thr Leu Gly1 5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser
Ser Lys Ser Leu Leu Tyr Lys 20 25 30Asp Gly Lys Thr Tyr Leu Asn Trp
Phe Gln Gln Arg Pro Gly Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu
Met Ser Thr Arg Ala Ser Gly Val Ser 50 55 60Asp Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu
Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Leu 85 90 95Val Glu Tyr
Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
11025112PRTArtificial Sequence2A10 VL humanised construct L18 25Asp
Ile Val Met Thr Gln Asp Pro Leu Ser Asn Pro Val Thr Leu Gly1 5 10
15Gln Pro Val Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Lys
20 25 30Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln
Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser Gly
Val Ser 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
Tyr Cys Gln Gln Leu 85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly
Thr Lys Leu Glu Ile Lys 100 105 11026462PRTArtificial Sequence2A10
heavy chain humanised construct H5 26Met Gly Trp Ser Cys Ile Ile
Leu Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln
Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ala Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu 50 55 60Glu Trp Met
Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu
Lys Phe Lys Ser Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser 85 90
95Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
100 105 110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr
Leu Val 115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
Phe Pro Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
Ala Ala Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215
220Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
Thr225 230 235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala
Pro Ser Val Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330
335Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
340 345 350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
Pro Pro 355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Thr Cys Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46027462PRTArtificial Sequence2A10 heavy chain humanised construct
H6 27Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Arg Ala Thr Met
Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46028462PRTArtificial Sequence2A10 heavy
chain humanised construct H700 28Met Gly Trp Ser Cys Ile Ile Leu
Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu
Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ala Ser Val Lys
Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met
His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly
Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys
Phe Lys Ser Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser 85 90 95Thr
Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105
110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230
235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val
Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345
350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46029462PRTArtificial Sequence2A10 heavy chain humanised construct
H14 29Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Val Lys 20 25 30Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Lys Gln Ala Pro
Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Arg Ala Thr Leu
Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46030462PRTArtificial Sequence2A10 heavy
chain humanised construct H15 30Met Gly Trp Ser Cys Ile Ile Leu Phe
Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Val Lys 20 25 30Pro Gly Ala Ser Val Lys Leu
Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His
Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn
Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe
Lys Ser Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser 85 90 95Thr Ala
Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Ser Ala Val 100 105
110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230
235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala
Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val
Val Asp Val Ser His Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315
320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile
Ala Val Glu Trp Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440
445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46031462PRTArtificial Sequence2A10 heavy chain humanised construct
H16 31Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Lys Gln Arg Pro
Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Lys Ala Thr Leu
Thr Val Asp Lys Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46032462PRTArtificial Sequence2A10 heavy
chain humanised construct H17 32Met Gly Trp Ser Cys Ile Ile Leu Phe
Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Val Lys 20 25 30Pro Gly Ala Ser Val Lys Leu
Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His
Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn
Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe
Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Thr Ser 85 90 95Thr Ala
Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105
110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230
235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val
Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345
350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46033462PRTArtificial Sequence2A10 heavy chain humanised construct
H18 33Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Val Lys 20 25 30Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Lys Gln Arg Pro
Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Lys Ala Thr Leu
Thr Val Asp Lys Ser Ser Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Ser Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46034238PRTArtificial Sequence2A10 light
chain humanised construct L6 34Met Gly Trp Ser Cys Ile Ile Leu Phe
Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Asp Ile Val Met Thr
Gln Ser Pro Leu Ser Leu Pro Val 20 25 30Thr Pro Gly Glu Pro Ala Ser
Ile Ser Cys Arg Ser Ser Lys Ser Leu 35 40 45Leu Tyr Lys Asp Gly Lys
Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro 50 55 60Gly Gln Ser Pro Gln
Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser65 70 75 80Gly Val Pro
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95Leu Lys
Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys 100 105
110Gln Gln Leu Val Glu Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val
115 120 125Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
Pro Pro 130 135 140Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val
Val Cys Leu Leu145 150 155 160Asn Asn Phe Tyr Pro Arg Glu Ala Lys
Val Gln Trp Lys Val Asp Asn 165 170 175Ala Leu Gln Ser Gly Asn Ser
Gln Glu Ser Val Thr Glu Gln Asp Ser 180 185 190Lys Asp Ser Thr Tyr
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala 195 200 205Asp Tyr Glu
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly 210 215 220Leu
Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230
23535238PRTArtificial Sequence2A10 light chain humanised construct
L13 35Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu
Pro Val 20 25 30Thr Leu Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser
Lys Ser Leu 35 40 45Leu Tyr Lys Asp Gly Lys Thr Tyr Leu Asn Trp Phe
Gln Gln Arg Pro 50 55 60Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Met
Ser Thr Arg Ala Ser65 70 75 80Gly Val Pro Asp Arg Phe Ser Gly Gly
Gly Ser Gly Thr Asp Phe Thr 85 90 95Leu Lys Ile Ser Arg Val Glu Ala
Glu Asp Val Gly Val Tyr Tyr Cys 100 105 110Gln Gln Leu Val Glu Tyr
Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu 115 120 125Glu Ile Lys Arg
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro 130 135 140Ser Asp
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu145 150 155
160Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
Asp Ser 180 185 190Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr
Leu Ser Lys Ala 195 200 205Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
Glu Val Thr His Gln Gly 210 215 220Leu Ser Ser Pro Val Thr Lys Ser
Phe Asn Arg Gly Glu Cys225 230 23536238PRTArtificial Sequence2A10
light chain humanised construct L14 36Met Gly Trp Ser Cys Ile Ile
Leu Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Asp Ile Val
Met Thr Gln Ser Pro Leu Ser Leu Pro Val 20 25 30Thr Leu Gly Gln Pro
Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu 35 40 45Leu Tyr Lys Asp
Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro 50 55 60Gly Gln Ser
Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser65 70 75 80Gly
Val Pro Asp Arg Phe Ser Gly Gly Gly Ser Gly Thr Asp Phe Thr 85 90
95Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys
100 105 110Gln Gln Leu Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly Thr
Lys Leu 115 120 125Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro 130 135 140Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu145 150 155 160Asn Asn Phe Tyr Pro Arg Glu
Ala Lys Val Gln Trp Lys Val Asp Asn 165 170 175Ala Leu Gln Ser Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser 180 185 190Lys Asp Ser
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala 195 200 205Asp
Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly 210 215
220Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230
23537238PRTArtificial Sequence2A10 light chain humanised construct
L15 37Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu
Pro Val 20 25 30Thr Leu Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser
Lys Ser Leu 35 40 45Leu Tyr Lys Asp Gly Lys Thr Tyr Leu Asn Trp Phe
Leu Gln Arg Pro 50 55 60Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Met
Ser Thr Arg Ala Ser65 70 75 80Gly Val Ser Asp Arg Phe Ser Gly Gly
Gly Ser Gly Thr Asp Phe Thr 85 90 95Leu Lys Ile Ser Arg Val
Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys 100 105 110Gln Gln Leu Val
Glu Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu 115 120 125Glu Ile
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro 130 135
140Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
Leu145 150 155 160Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp
Lys Val Asp Asn 165 170 175Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
Val Thr Glu Gln Asp Ser 180 185 190Lys Asp Ser Thr Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala 195 200 205Asp Tyr Glu Lys His Lys
Val Tyr Ala Cys Glu Val Thr His Gln Gly 210 215 220Leu Ser Ser Pro
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230
23538238PRTArtificial Sequence2A10 light chain humanised construct
L16 38Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Asp Ile Val Met Thr Gln Ser Pro Leu Ser Asn
Pro Val 20 25 30Thr Leu Gly Gln Pro Val Ser Ile Ser Cys Arg Ser Ser
Lys Ser Leu 35 40 45Leu Tyr Lys Asp Gly Lys Thr Tyr Leu Asn Trp Phe
Leu Gln Arg Pro 50 55 60Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Met
Ser Thr Arg Ala Ser65 70 75 80Gly Val Pro Asp Arg Phe Ser Gly Gly
Gly Ser Gly Thr Asp Phe Thr 85 90 95Leu Lys Ile Ser Arg Val Glu Ala
Glu Asp Val Gly Val Tyr Tyr Cys 100 105 110Gln Gln Leu Val Glu Tyr
Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu 115 120 125Glu Ile Lys Arg
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro 130 135 140Ser Asp
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu145 150 155
160Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
Asp Ser 180 185 190Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr
Leu Ser Lys Ala 195 200 205Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
Glu Val Thr His Gln Gly 210 215 220Leu Ser Ser Pro Val Thr Lys Ser
Phe Asn Arg Gly Glu Cys225 230 23539238PRTArtificial Sequence2A10
light chain humanised construct L17 39Met Gly Trp Ser Cys Ile Ile
Leu Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Asp Ile Val
Met Thr Gln Asp Pro Leu Ser Leu Pro Val 20 25 30Thr Leu Gly Gln Pro
Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu 35 40 45Leu Tyr Lys Asp
Gly Lys Thr Tyr Leu Asn Trp Phe Gln Gln Arg Pro 50 55 60Gly Gln Ser
Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser65 70 75 80Gly
Val Ser Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90
95Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys
100 105 110Gln Gln Leu Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly Thr
Lys Leu 115 120 125Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro 130 135 140Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu145 150 155 160Asn Asn Phe Tyr Pro Arg Glu
Ala Lys Val Gln Trp Lys Val Asp Asn 165 170 175Ala Leu Gln Ser Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser 180 185 190Lys Asp Ser
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala 195 200 205Asp
Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly 210 215
220Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230
23540238PRTArtificial Sequence2A10 light chain humanised construct
L18 40Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Asp Ile Val Met Thr Gln Asp Pro Leu Ser Asn
Pro Val 20 25 30Thr Leu Gly Gln Pro Val Ser Ile Ser Cys Arg Ser Ser
Lys Ser Leu 35 40 45Leu Tyr Lys Asp Gly Lys Thr Tyr Leu Asn Trp Phe
Leu Gln Arg Pro 50 55 60Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Met
Ser Thr Arg Ala Ser65 70 75 80Gly Val Ser Asp Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe Thr 85 90 95Leu Lys Ile Ser Arg Val Glu Ala
Glu Asp Val Gly Val Tyr Tyr Cys 100 105 110Gln Gln Leu Val Glu Tyr
Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu 115 120 125Glu Ile Lys Arg
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro 130 135 140Ser Asp
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu145 150 155
160Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
Asp Ser 180 185 190Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr
Leu Ser Lys Ala 195 200 205Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
Glu Val Thr His Gln Gly 210 215 220Leu Ser Ser Pro Val Thr Lys Ser
Phe Asn Arg Gly Glu Cys225 230 23541339DNAMurine 41caggtccaac
tgcagcagcc tgggactgaa ctggtgaagc ctggggcttc agtgaagctg 60tcctgcaagg
cttctggcta caccttcacc agctactgga tgcactgggt gaagcagagg
120cctggacaag gccttgagtg gattggaaat attaatccta gcaatggtgg
tactaactac 180aatgagaagt tcaagagcaa ggccacactg actgtagaca
aatcctccag cacagcctac 240atgcagctca gcagcctgac atctgaggac
tctgcggtct attattgtga actgggacag 300ggctactggg gccaaggcac
cactctcaca gtctcctca 33942336DNAMurine 42gatattgtga taacccagga
tgaactctcc aatcctgtca cttctggaga atcagtttcc 60atctcctgca ggtctagtaa
gagtctccta tataaggatg ggaagacata cttgaattgg 120tttctgcaga
gaccaggaca atctcctcag ctcctgatct atttgatgtc cacccgtgca
180tcaggagtct cagaccggtt tagtggcagt gggtcaggaa cagatttcac
cctggaaatc 240agtagagtga aggctgagga tgtgggtgtg tattactgtc
aacaacttgt agagtatccg 300ctcacgttcg gtgctgggac caagctggag ctgaaa
336431389DNAArtificial SequencePN encoding Chimeric heavy chain Hc
SEQ ID 9 43atgggatgga gctgtatcat cctctttttg gtagcagcag ctacaggtgt
ccactcccag 60gtccaactgc agcagcctgg gactgaactg gtgaagcctg gggcttcagt
gaagctgtcc 120tgcaaggctt ctggctacac cttcaccagc tactggatgc
actgggtgaa gcagaggcct 180ggacaaggcc ttgagtggat tggaaatatt
aatcctagca atggtggtac taactacaat 240gagaagttca agagcaaggc
cacactgact gtagacaaat cctccagcac agcctacatg 300cagctcagca
gcctgacatc tgaggactct gcggtctatt attgtgaact gggacagggc
360tactggggcc aaggcacact agtcacagtc tcctcagcct ccaccaaggg
cccatcggtc 420ttccccctgg caccctcctc caagagcacc tctgggggca
cagcggccct gggctgcctg 480gtcaaggact acttccccga accggtgacg
gtgtcgtgga actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc
tgtcctacag tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc
cctccagcag cttgggcacc cagacctaca tctgcaacgt gaatcacaag
660cccagcaaca ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa
aactcacaca 720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt
cagtcttcct cttcccccca 780aaacccaagg acaccctcat gatctcccgg
acccctgagg tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga
ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga
caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc
960ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
ggtctccaac 1020aaagccctcc cagcccccat cgagaaaacc atctccaaag
ccaaagggca gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg
gatgagctga ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt
ctatcccagc gacatcgccg tggagtggga gagcaatggg 1200cagccggaga
acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc
1260ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
cttctcatgc 1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga
agagcctctc cctgtctccg 1380ggtaaatga 138944720DNAArtificial
SequencePN encoding Chimeric light chain Lc SEQ ID 10 44atgaggtgct
ctcttcagtt tctgggggtg cttatgttct ggatctctgg agtcagtggg 60gatattgtga
taacccagga tgaactctcc aatcctgtca cttctggaga atcagtttcc
120atctcctgca ggtctagtaa gagtctccta tataaggatg ggaagacata
cttgaattgg 180tttctgcaga gaccaggaca atctcctcag ctcctgatct
atttgatgtc cacccgtgca 240tcaggagtct cagaccggtt tagtggcagt
gggtcaggaa cagatttcac cctggaaatc 300agtagagtga aggctgagga
tgtgggtgtg tattactgtc aacaacttgt agagtatccg 360ctcacgttcg
gtgctgggac caagctggag ctgaaacgta cggtggctgc accatctgtc
420ttcatcttcc cgccatctga tgagcagttg aaatctggaa ctgcctctgt
tgtgtgcctg 480ctgaataact tctatcccag agaggccaaa gtacagtgga
aggtggacaa cgccctccaa 540tcgggtaact cccaggagag tgtcacagag
caggacagca aggacagcac ctacagcctc 600agcagcaccc tgacgctgag
caaagcagac tacgagaaac acaaagtcta cgcctgcgaa 660gtcacccatc
agggcctgag ctcgcccgtc acaaagagct tcaacagggg agagtgttag
72045339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H5 SEQ ID 11 45caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gcgacaggcc 120cctggacaag ggcttgagtg
gatgggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcag agtcaccatg accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33946339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H6 SEQ ID 12 46caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gcgacaggcc 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcag agccaccatg accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33947339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H700 SEQ ID 13 47caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gcgacaggcc 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcag agccaccctg accgtagaca aatccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33948339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H14 SEQ ID 14 48caggtgcagc tggtgcagtc tggggctgag
gtggtcaagc ctggggcctc agtgaagctc 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaaacaggcc 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcag agccaccctc accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33949339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H15 SEQ ID 15 49caggtgcagc tggtgcagtc tggggctgag
gtggtcaagc ctggggcctc agtgaagctc 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaaacagcga 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa agccaccctc accgtcgaca cgtccagcag cacagcctac
240atggagctga gcagcctgag atctgaggac agcgccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33950339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H16 SEQ ID 16 50caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaaacagcga 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa agccaccctc accgtcgaca aatccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33951339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H17 SEQ ID 17 51caggtgcagc tggtgcagtc tggggctgag
gtggtcaagc ctggggcctc agtgaagctc 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaaacagcga 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa agccaccctc accgtcgaca aatccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33952339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H18 SEQ ID 18 52caggtgcagc tggtgcagtc tggggctgag
gtggtcaagc ctggggcctc agtgaagctc 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaaacagcga 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa agccaccctc accgtcgaca aatccagcag cacagcctac
240atggagctga gcagcctgag atctgaggac agcgccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
33953336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L6 SEQ ID 19 53gatattgtga tgactcagtc tccactctcc
ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120ttcctgcagc gaccagggca
gtctccacag ctcctgatct atttgatgtc cacccgtgca 180tcaggggtcc
ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc
240agcagagtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgttcg gcggagggac caaggtggag atcaaa
33654336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L13 SEQ ID 20 54gatattgtga tgacccagtc tccactctcc
ctgcccgtca cccttggaca gccggcctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttcagcaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtcc
cagacagatt cagcggcggt gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
33655336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L14 SEQ ID 21 55gatattgtga tgacccagtc tccactctcc
ctgcccgtca cccttggaca gccggcctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttctccaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtcc
cagacagatt cagcggcggt gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
33656336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L15 SEQ ID 22 56gatattgtga tgacccagtc tccactctcc
ctgcccgtca cccttggaca gccggcctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttctccaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtca
gcgacagatt cagcggcggt gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
33657336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L16 SEQ ID 23 57gatattgtga tgacccagtc tccactctcc
aaccccgtca cccttggaca gccggtctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttctccaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtcc
cagacagatt cagcggcggt gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
33658336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L17 SEQ ID 24 58gatattgtga tgacccagga cccactctcc
ctgcccgtca cccttggaca gccggcctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttcagcaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtca
gcgacagatt cagcggcagc gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
33659336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L18 SEQ ID 25 59gatattgtga tgacccagga cccactctcc
aaccccgtca cccttggaca gccggtctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttctccaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtca
gcgacagatt cagcggcagc gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
336601389DNAArtificial SequencePN encoding 2A10 heavy chain
humanised construct H5 SEQ ID 26 60atgggatgga gctgtatcat
cctcttcttg
gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg tgcagtctgg ggctgaggtg
aagaagcctg gggcctcagt gaaggtttcc 120tgcaaggcat ctggatacac
cttcaccagc tactggatgc actgggtgcg acaggcccct 180ggacaagggc
ttgagtggat gggaaatatt aatcctagca atggtggtac taactacaat
240gagaagttca agagcagagt caccatgacc agggacacgt ccacgagcac
agcctacatg 300gagctgagca gcctgagatc tgaggacacg gccgtgtatt
actgtgaact gggacagggc 360tactggggcc agggaacact agtcacagtc
tcctcagcct ccaccaaggg cccatcggtc 420ttccccctgg caccctcctc
caagagcacc tctgggggca cagcggccct gggctgcctg 480gtcaaggact
acttccccga accggtgacg gtgtcgtgga actcaggcgc cctgaccagc
540ggcgtgcaca ccttcccggc tgtcctacag tcctcaggac tctactccct
cagcagcgtg 600gtgaccgtgc cctccagcag cttgggcacc cagacctaca
tctgcaacgt gaatcacaag 660cccagcaaca ccaaggtgga caagaaagtt
gagcccaaat cttgtgacaa aactcacaca 720tgcccaccgt gcccagcacc
tgaactcgcg ggggcaccgt cagtcttcct cttcccccca 780aaacccaagg
acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac
840gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
ggaggtgcat 900aatgccaaga caaagccgcg ggaggagcag tacaacagca
cgtaccgtgt ggtcagcgtc 960ctcaccgtcc tgcaccagga ctggctgaat
ggcaaggagt acaagtgcaa ggtctccaac 1020aaagccctcc cagcccccat
cgagaaaacc atctccaaag ccaaagggca gccccgagaa 1080ccacaggtgt
acaccctgcc cccatcccgg gatgagctga ccaagaacca ggtcagcctg
1140acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
gagcaatggg 1200cagccggaga acaactacaa gaccacgcct cccgtgctgg
actccgacgg ctccttcttc 1260ctctacagca agctcaccgt ggacaagagc
aggtggcagc aggggaacgt cttctcatgc 1320tccgtgatgc atgaggctct
gcacaaccac tacacgcaga agagcctctc cctgtctccg 1380ggtaaatga
1389611389DNAArtificial SequencePN encoding 2A10 heavy chain
humanised construct H6 SEQ ID 27 61atgggatgga gctgtatcat cctcttcttg
gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg tgcagtctgg ggctgaggtg
aagaagcctg gggcctcagt gaaggtttcc 120tgcaaggcat ctggatacac
cttcaccagc tactggatgc actgggtgcg acaggcccct 180ggacaagggc
ttgagtggat cggaaatatt aatcctagca atggtggtac taactacaat
240gagaagttca agagcagagc caccatgacc agggacacgt ccacgagcac
agcctacatg 300gagctgagca gcctgagatc tgaggacacg gccgtgtatt
actgtgaact gggacagggc 360tactggggcc agggaacact agtcacagtc
tcctcagcct ccaccaaggg cccatcggtc 420ttccccctgg caccctcctc
caagagcacc tctgggggca cagcggccct gggctgcctg 480gtcaaggact
acttccccga accggtgacg gtgtcgtgga actcaggcgc cctgaccagc
540ggcgtgcaca ccttcccggc tgtcctacag tcctcaggac tctactccct
cagcagcgtg 600gtgaccgtgc cctccagcag cttgggcacc cagacctaca
tctgcaacgt gaatcacaag 660cccagcaaca ccaaggtgga caagaaagtt
gagcccaaat cttgtgacaa aactcacaca 720tgcccaccgt gcccagcacc
tgaactcgcg ggggcaccgt cagtcttcct cttcccccca 780aaacccaagg
acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac
840gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
ggaggtgcat 900aatgccaaga caaagccgcg ggaggagcag tacaacagca
cgtaccgtgt ggtcagcgtc 960ctcaccgtcc tgcaccagga ctggctgaat
ggcaaggagt acaagtgcaa ggtctccaac 1020aaagccctcc cagcccccat
cgagaaaacc atctccaaag ccaaagggca gccccgagaa 1080ccacaggtgt
acaccctgcc cccatcccgg gatgagctga ccaagaacca ggtcagcctg
1140acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
gagcaatggg 1200cagccggaga acaactacaa gaccacgcct cccgtgctgg
actccgacgg ctccttcttc 1260ctctacagca agctcaccgt ggacaagagc
aggtggcagc aggggaacgt cttctcatgc 1320tccgtgatgc atgaggctct
gcacaaccac tacacgcaga agagcctctc cctgtctccg 1380ggtaaatga
1389621389DNAArtificial SequencePN encoding 2A10 heavy chain
humanised construct H700 SEQ ID 28 62atgggatgga gctgtatcat
cctcttcttg gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg tgcagtctgg
ggctgaggtg aagaagcctg gggcctcagt gaaggtttcc 120tgcaaggcat
ctggatacac cttcaccagc tactggatgc actgggtgcg acaggcccct
180ggacaagggc ttgagtggat cggaaatatt aatcctagca atggtggtac
taactacaat 240gagaagttca agagcagagc caccctgacc gtagacaaat
ccacgagcac agcctacatg 300gagctgagca gcctgagatc tgaggacacg
gccgtgtatt actgtgaact gggacagggc 360tactggggcc agggaacact
agtcacagtc tcctcagcct ccaccaaggg cccatcggtc 420ttccccctgg
caccctcctc caagagcacc tctgggggca cagcggccct gggctgcctg
480gtcaaggact acttccccga accggtgacg gtgtcgtgga actcaggcgc
cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag tcctcaggac
tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag cttgggcacc
cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca ccaaggtgga
caagaaagtt gagcccaaat cttgtgacaa aactcacaca 720tgcccaccgt
gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct cttcccccca
780aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc aactggtacg
tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg ggaggagcag
tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc tgcaccagga
ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac 1020aaagccctcc
cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa
1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga ccaagaacca
ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc gacatcgccg
tggagtggga gagcaatggg 1200cagccggaga acaactacaa gaccacgcct
cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca agctcaccgt
ggacaagagc aggtggcagc aggggaacgt cttctcatgc 1320tccgtgatgc
atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg
1380ggtaaatga 1389631389DNAArtificial SequencePN encoding 2A10
heavy chain humanised construct H14 SEQ ID 29 63atgggatgga
gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg
tgcagtctgg ggctgaggtg gtcaagcctg gggcctcagt gaagctctcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgaa
acaggcccct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcagagc caccctcacc
agggacacgt ccacgagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacacg gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct ccaccaaggg cccatcggtc
420ttccccctgg caccctcctc caagagcacc tctgggggca cagcggccct
gggctgcctg 480gtcaaggact acttccccga accggtgacg gtgtcgtgga
actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag
tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag
cttgggcacc cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca
ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactcacaca
720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct
cttcccccca 780aaacccaagg acaccctcat gatctcccgg acccctgagg
tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc
aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg
ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc
tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac
1020aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga
ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc
gacatcgccg tggagtggga gagcaatggg 1200cagccggaga acaactacaa
gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca
agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc
1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
cctgtctccg 1380ggtaaatga 1389641389DNAArtificial SequencePN
encoding 2A10 heavy chain humanised construct H15 SEQ ID 30
64atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag
60gtgcagctgg tgcagtctgg ggctgaggtg gtcaagcctg gggcctcagt gaagctctcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgaa
acagcgacct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcaaagc caccctcacc
gtcgacacgt ccagcagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacagc gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct ccaccaaggg cccatcggtc
420ttccccctgg caccctcctc caagagcacc tctgggggca cagcggccct
gggctgcctg 480gtcaaggact acttccccga accggtgacg gtgtcgtgga
actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag
tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag
cttgggcacc cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca
ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactcacaca
720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct
cttcccccca 780aaacccaagg acaccctcat gatctcccgg acccctgagg
tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc
aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg
ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc
tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac
1020aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga
ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc
gacatcgccg tggagtggga gagcaatggg 1200cagccggaga acaactacaa
gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca
agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc
1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
cctgtctccg 1380ggtaaatga 1389651389DNAArtificial SequencePN
encoding 2A10 heavy chain humanised construct H16 SEQ ID 31
65atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag
60gtgcagctgg tgcagtctgg ggctgaggtg aagaagcctg gggcctcagt gaaggtttcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgaa
acagcgacct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcaaagc caccctcacc
gtcgacaaat ccacgagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacacg gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct ccaccaaggg cccatcggtc
420ttccccctgg caccctcctc caagagcacc tctgggggca cagcggccct
gggctgcctg 480gtcaaggact acttccccga accggtgacg gtgtcgtgga
actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag
tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag
cttgggcacc cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca
ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactcacaca
720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct
cttcccccca 780aaacccaagg acaccctcat gatctcccgg acccctgagg
tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc
aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg
ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc
tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac
1020aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga
ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc
gacatcgccg tggagtggga gagcaatggg 1200cagccggaga acaactacaa
gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca
agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc
1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
cctgtctccg 1380ggtaaatga 1389661389DNAArtificial SequencePN
encoding 2A10 heavy chain humanised construct H17 SEQ ID 32
66atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag
60gtgcagctgg tgcagtctgg ggctgaggtg gtcaagcctg gggcctcagt gaagctctcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgaa
acagcgacct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcaaagc caccctcacc
gtcgacaaat ccacgagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacacg gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct ccaccaaggg cccatcggtc
420ttccccctgg caccctcctc caagagcacc tctgggggca cagcggccct
gggctgcctg 480gtcaaggact acttccccga accggtgacg gtgtcgtgga
actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag
tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag
cttgggcacc cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca
ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactcacaca
720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct
cttcccccca 780aaacccaagg acaccctcat gatctcccgg acccctgagg
tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc
aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg
ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc
tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac
1020aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga
ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc
gacatcgccg tggagtggga gagcaatggg 1200cagccggaga acaactacaa
gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca
agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc
1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
cctgtctccg 1380ggtaaatga 1389671389DNAArtificial SequencePN
encoding 2A10 heavy chain humanised construct H18 SEQ ID 33
67atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag
60gtgcagctgg tgcagtctgg ggctgaggtg gtcaagcctg gggcctcagt gaagctctcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgaa
acagcgacct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcaaagc caccctcacc
gtcgacaaat ccagcagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacagc gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct ccaccaaggg cccatcggtc
420ttccccctgg caccctcctc caagagcacc tctgggggca cagcggccct
gggctgcctg 480gtcaaggact acttccccga accggtgacg gtgtcgtgga
actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag
tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag
cttgggcacc cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca
ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactcacaca
720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct
cttcccccca 780aaacccaagg acaccctcat gatctcccgg acccctgagg
tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc
aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg
ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc
tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac
1020aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga
ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc
gacatcgccg tggagtggga gagcaatggg 1200cagccggaga acaactacaa
gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca
agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc
1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
cctgtctccg 1380ggtaaatga 138968717DNAArtificial SequencePN encoding
2A10 light chain humanised construct L6 SEQ ID 34 68atgggatgga
gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactccgat 60attgtgatga
ctcagtctcc actctccctg cccgtcaccc ctggagagcc ggcctccatc
120tcctgcaggt ctagtaagag tctcctatat aaggatggga agacatactt
gaattggttc 180ctgcagcgac cagggcagtc tccacagctc ctgatctatt
tgatgtccac ccgtgcatca 240ggggtccctg acaggttcag tggcagtgga
tcaggcacag attttacact gaaaatcagc 300agagtggagg ctgaggatgt
tggggtttat tactgccaac aacttgtaga gtatccgctc 360acgttcggcg
gagggaccaa ggtggagatc aaacgtacgg tggctgcacc atctgtcttc
420atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt
gtgcctgctg 480aataacttct atcccagaga ggccaaagta cagtggaagg
tggacaacgc cctccaatcg 540ggtaactccc aggagagtgt cacagagcag
gacagcaagg acagcaccta cagcctcagc 600agcaccctga cgctgagcaa
agcagactac gagaaacaca aagtctacgc ctgcgaagtc 660acccatcagg
gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgttag
71769717DNAArtificial SequencePN encoding 2A10 light chain
humanised construct L13 SEQ ID 35 69atgggatgga gctgtatcat
cctcttcttg gtagcaacag ctacaggtgt ccactccgat 60attgtgatga cccagtctcc
actctccctg cccgtcaccc ttggacagcc ggcctccatc 120tcctgcaggt
ctagtaagag tctcctatat aaggatggga agacatactt gaattggttt
180cagcagaggc caggccaatc tccacagctc ctaatttatt tgatgtccac
ccgtgcatct 240ggggtcccag acagattcag cggcggtggg tcaggcactg
atttcacact gaaaatcagc 300agggtggagg ctgaggatgt tggggtttat
tactgccaac aacttgtaga gtatccgctc 360acgtttggcc aggggaccaa
gctggagatc aaacgtacgg tggctgcacc atctgtcttc 420atcttcccgc
catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg
480aataacttct atcccagaga ggccaaagta cagtggaagg tggacaacgc
cctccaatcg 540ggtaactccc aggagagtgt cacagagcag gacagcaagg
acagcaccta cagcctcagc 600agcaccctga cgctgagcaa agcagactac
gagaaacaca aagtctacgc ctgcgaagtc 660acccatcagg gcctgagctc
gcccgtcaca aagagcttca acaggggaga gtgttag 71770717DNAArtificial
SequencePN encoding 2A10 light chain humanised construct L14 SEQ ID
36 70atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt
ccactccgat 60attgtgatga cccagtctcc actctccctg cccgtcaccc ttggacagcc
ggcctccatc 120tcctgcaggt ctagtaagag tctcctatat aaggatggga
agacatactt gaattggttt 180ctccagaggc caggccaatc tccacagctc
ctaatttatt tgatgtccac ccgtgcatct 240ggggtcccag acagattcag
cggcggtggg tcaggcactg atttcacact gaaaatcagc 300agggtggagg
ctgaggatgt tggggtttat tactgccaac aacttgtaga gtatccgctc
360acgtttggcc aggggaccaa gctggagatc aaacgtacgg tggctgcacc
atctgtcttc 420atcttcccgc catctgatga gcagttgaaa tctggaactg
cctctgttgt gtgcctgctg 480aataacttct atcccagaga ggccaaagta
cagtggaagg tggacaacgc cctccaatcg 540ggtaactccc aggagagtgt
cacagagcag gacagcaagg acagcaccta cagcctcagc 600agcaccctga
cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc
660acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgttag
71771717DNAArtificial SequencePN encoding 2A10 light chain
humanised construct L15 SEQ ID 37 71atgggatgga gctgtatcat
cctcttcttg gtagcaacag ctacaggtgt ccactccgat 60attgtgatga cccagtctcc
actctccctg cccgtcaccc ttggacagcc ggcctccatc 120tcctgcaggt
ctagtaagag tctcctatat aaggatggga agacatactt gaattggttt
180ctccagaggc caggccaatc tccacagctc ctaatttatt tgatgtccac
ccgtgcatct 240ggggtcagcg acagattcag cggcggtggg tcaggcactg
atttcacact gaaaatcagc 300agggtggagg ctgaggatgt tggggtttat
tactgccaac aacttgtaga gtatccgctc 360acgtttggcc aggggaccaa
gctggagatc aaacgtacgg tggctgcacc atctgtcttc 420atcttcccgc
catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg
480aataacttct atcccagaga ggccaaagta cagtggaagg tggacaacgc
cctccaatcg 540ggtaactccc aggagagtgt cacagagcag gacagcaagg
acagcaccta cagcctcagc 600agcaccctga cgctgagcaa agcagactac
gagaaacaca aagtctacgc ctgcgaagtc 660acccatcagg gcctgagctc
gcccgtcaca aagagcttca acaggggaga gtgttag 71772717DNAArtificial
SequencePN encoding 2A10 light chain humanised construct L16 SEQ ID
38 72atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt
ccactccgat 60attgtgatga cccagtctcc actctccaac cccgtcaccc ttggacagcc
ggtctccatc 120tcctgcaggt ctagtaagag tctcctatat aaggatggga
agacatactt gaattggttt 180ctccagaggc caggccaatc tccacagctc
ctaatttatt tgatgtccac ccgtgcatct 240ggggtcccag acagattcag
cggcggtggg tcaggcactg atttcacact gaaaatcagc 300agggtggagg
ctgaggatgt tggggtttat tactgccaac aacttgtaga gtatccgctc
360acgtttggcc aggggaccaa gctggagatc aaacgtacgg tggctgcacc
atctgtcttc 420atcttcccgc catctgatga gcagttgaaa tctggaactg
cctctgttgt gtgcctgctg 480aataacttct atcccagaga ggccaaagta
cagtggaagg tggacaacgc cctccaatcg 540ggtaactccc aggagagtgt
cacagagcag gacagcaagg acagcaccta cagcctcagc 600agcaccctga
cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc
660acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgttag
71773717DNAArtificial SequencePN encoding 2A10 light chain
humanised construct L17 SEQ ID 39 73atgggatgga gctgtatcat
cctcttcttg gtagcaacag ctacaggtgt ccactccgat 60attgtgatga cccaggaccc
actctccctg cccgtcaccc ttggacagcc ggcctccatc 120tcctgcaggt
ctagtaagag tctcctatat aaggatggga agacatactt gaattggttt
180cagcagaggc caggccaatc tccacagctc ctaatttatt tgatgtccac
ccgtgcatct 240ggggtcagcg acagattcag cggcagcggg tcaggcactg
atttcacact gaaaatcagc 300agggtggagg ctgaggatgt tggggtttat
tactgccaac aacttgtaga gtatccgctc 360acgtttggcc aggggaccaa
gctggagatc aaacgtacgg tggctgcacc atctgtcttc 420atcttcccgc
catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg
480aataacttct atcccagaga ggccaaagta cagtggaagg tggacaacgc
cctccaatcg 540ggtaactccc aggagagtgt cacagagcag gacagcaagg
acagcaccta cagcctcagc 600agcaccctga cgctgagcaa agcagactac
gagaaacaca aagtctacgc ctgcgaagtc 660acccatcagg gcctgagctc
gcccgtcaca aagagcttca acaggggaga gtgttag 71774717DNAArtificial
SequencePN encoding 2A10 light chain humanised construct L18 SEQ ID
40 74atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt
ccactccgat 60attgtgatga cccaggaccc actctccaac cccgtcaccc ttggacagcc
ggtctccatc 120tcctgcaggt ctagtaagag tctcctatat aaggatggga
agacatactt gaattggttt 180ctccagaggc caggccaatc tccacagctc
ctaatttatt tgatgtccac ccgtgcatct 240ggggtcagcg acagattcag
cggcagcggg tcaggcactg atttcacact gaaaatcagc 300agggtggagg
ctgaggatgt tggggtttat tactgccaac aacttgtaga gtatccgctc
360acgtttggcc aggggaccaa gctggagatc aaacgtacgg tggctgcacc
atctgtcttc 420atcttcccgc catctgatga gcagttgaaa tctggaactg
cctctgttgt gtgcctgctg 480aataacttct atcccagaga ggccaaagta
cagtggaagg tggacaacgc cctccaatcg 540ggtaactccc aggagagtgt
cacagagcag gacagcaagg acagcaccta cagcctcagc 600agcaccctga
cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc
660acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgttag
7177519PRTArtificial SequenceCampath leader sequence 75Met Gly Trp
Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His
Ser76201PRTArtificial SequenceAmino acids 586-785 of human NOGO A
(NOGO-A56) 76Met Gln Glu Ser Leu Tyr Pro Ala Ala Gln Leu Cys Pro
Ser Phe Glu1 5 10 15Glu Ser Glu Ala Thr Pro Ser Pro Val Leu Pro Asp
Ile Val Met Glu 20 25 30Ala Pro Leu Asn Ser Ala Val Pro Ser Ala Gly
Ala Ser Val Ile Gln 35 40 45Pro Ser Ser Ser Pro Leu Glu Ala Ser Ser
Val Asn Tyr Glu Ser Ile 50 55 60Lys His Glu Pro Glu Asn Pro Pro Pro
Tyr Glu Glu Ala Met Ser Val65 70 75 80Ser Leu Lys Lys Val Ser Gly
Ile Lys Glu Glu Ile Lys Glu Pro Glu 85 90 95Asn Ile Asn Ala Ala Leu
Gln Glu Thr Glu Ala Pro Tyr Ile Ser Ile 100 105 110Ala Cys Asp Leu
Ile Lys Glu Thr Lys Leu Ser Ala Glu Pro Ala Pro 115 120 125Asp Phe
Ser Asp Tyr Ser Glu Met Ala Lys Val Glu Gln Pro Val Pro 130 135
140Asp His Ser Glu Leu Val Glu Asp Ser Ser Pro Asp Ser Glu Pro
Val145 150 155 160Asp Leu Phe Ser Asp Asp Ser Ile Pro Asp Val Pro
Gln Lys Gln Asp 165 170 175Glu Thr Val Met Leu Val Lys Glu Ser Leu
Thr Glu Thr Ser Phe Glu 180 185 190Ser Met Ile Glu Tyr Glu Asn Lys
Glu 195 20077113PRTArtificial Sequence2A10 VH humanised construct
H1 77Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly
Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Ser Tyr 20 25 30Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr
Asn Glu Lys Phe 50 55 60Lys Ser Arg Val Thr Met Thr Arg Asp Thr Ser
Thr Ser Thr Val Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly
Gln Gly Thr Leu Val Thr Val Ser 100 105 110Ser78112PRTArtificial
Sequence2A10 VL humanised construct L11 78Asp Ile Val Ile Thr Gln
Ser Pro Leu Ser Leu Pro Val Thr Leu Gly1 5 10 15Gln Pro Ala Ser Ile
Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Lys 20 25 30Asp Gly Lys Thr
Tyr Leu Asn Trp Phe Gln Gln Arg Pro Gly Gln Ser 35 40 45Pro Gln Leu
Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60Asp Arg
Phe Ser Gly Gly Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75
80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Leu
85 90 95Val Glu Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys 100 105 11079462PRTArtificial Sequence2A10 heavy chain
humanised construct H1 79Met Gly Trp Ser Cys Ile Ile Leu Phe Leu
Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu 50 55 60Glu Trp Met Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys
Ser Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Val Tyr
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr
Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120
125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His Thr Phe
Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser
Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230 235
240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe
245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360
365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 46080238PRTArtificial
Sequence2A10 light chain humanised construct L11 80Met Gly Trp Ser
Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser
Asp Ile Val Ile Thr Gln Ser Pro Leu Ser Leu Pro Val 20 25 30Thr Leu
Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu 35 40 45Leu
Tyr Lys Asp Gly Lys Thr Tyr Leu Asn Trp Phe Gln Gln Arg Pro 50 55
60Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Met Ser Thr Arg Ala Ser65
70 75 80Gly Val Pro Asp Arg Phe Ser Gly Gly Gly Ser Gly Thr Asp Phe
Thr 85 90 95Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
Tyr Cys 100 105 110Gln Gln Leu Val Glu Tyr Pro Leu Thr Phe Gly Gln
Gly Thr Lys Leu 115 120 125Glu Ile Lys Arg Thr Val Ala Ala Pro Ser
Val Phe Ile Phe Pro Pro 130 135 140Ser Asp Glu Gln Leu Lys Ser Gly
Thr Ala Ser Val Val Cys Leu Leu145 150 155 160Asn Asn Phe Tyr Pro
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn 165 170 175Ala Leu Gln
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser 180 185 190Lys
Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala 195 200
205Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
210 215 220Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
Cys225 230 23581339PRTArtificial SequencePN encoding 2A10 VH
humanised construct H1 SEQ ID 77 81Cys Ala Gly Gly Thr Gly Cys Ala
Gly Cys Thr Gly Gly Thr Gly Cys1 5 10 15Ala Gly Thr Cys Thr Gly Gly
Gly Gly Cys Thr Gly Ala Gly Gly Thr 20 25 30Gly Ala Ala Gly Ala Ala
Gly Cys Cys Thr Gly Gly Gly Gly Cys Cys 35 40 45Thr Cys Ala Gly Thr
Gly Ala Ala Gly Gly Thr Thr Thr Cys Cys Thr 50 55 60Gly Cys Ala Ala
Gly Gly Cys Ala Thr Cys Thr Gly Gly Ala Thr Ala65 70 75 80Cys Ala
Cys Cys Thr Thr Cys Ala Cys Cys Ala Gly Cys Thr Ala Cys 85 90 95Thr
Gly Gly Ala Thr Gly Cys Ala Cys Thr Gly Gly Gly Thr Gly Cys 100 105
110Gly Ala Cys Ala Gly Gly Cys Cys Cys Cys Thr Gly Gly Ala Cys Ala
115 120 125Ala Gly Gly Gly Cys Thr Thr Gly Ala Gly Thr Gly Gly Ala
Thr Gly 130 135 140Gly Gly Ala Ala Ala Thr Ala Thr Thr Ala Ala Thr
Cys Cys Thr Ala145 150 155 160Gly Cys Ala Ala Thr Gly Gly Thr Gly
Gly Thr Ala Cys Thr Ala Ala 165 170 175Cys Thr Ala Cys Ala Ala Thr
Gly Ala Gly Ala Ala Gly Thr Thr Cys 180 185 190Ala Ala Gly Ala Gly
Cys Ala Gly Ala Gly Thr Cys Ala Cys Cys Ala 195 200 205Thr Gly Ala
Cys Cys Ala Gly Gly Gly Ala Cys Ala Cys Gly Thr Cys 210 215 220Cys
Ala Cys Gly Ala Gly Cys Ala Cys Ala Gly Thr Cys Thr Ala Cys225 230
235 240Ala Thr Gly Gly Ala Gly Cys Thr Gly Ala Gly Cys Ala Gly Cys
Cys 245 250 255Thr Gly Ala Gly Ala Thr Cys Thr Gly Ala Gly Gly Ala
Cys Ala Cys 260 265 270Gly Gly Cys Cys Gly Thr Gly Thr Ala Thr Thr
Ala Cys Thr Gly Thr 275 280 285Gly Ala Ala Cys Thr Gly Gly Gly Ala
Cys Ala Gly Gly Gly Cys Thr 290 295 300Ala Cys Thr Gly Gly Gly Gly
Cys Cys Ala Gly Gly Gly Ala Ala Cys305 310 315 320Ala Cys Thr Ala
Gly Thr Cys Ala Cys Ala Gly Thr Cys Thr Cys Cys 325 330 335Thr Cys
Ala82336DNAArtificial SequencePN encoding 2A10 VL humanised
construct L11 SEQ ID 78 82gatattgtga taacccagtc tccactctcc
ctgcccgtca cccttggaca gccggcctcc 60atctcctgca ggtctagtaa gagtctccta
tataaggatg ggaagacata cttgaattgg 120tttcagcaga ggccaggcca
atctccacag ctcctaattt atttgatgtc cacccgtgca 180tctggggtcc
cagacagatt cagcggcggt gggtcaggca ctgatttcac actgaaaatc
240agcagggtgg aggctgagga tgttggggtt tattactgcc aacaacttgt
agagtatccg 300ctcacgtttg gccaggggac caagctggag atcaaa
336831389DNAArtificial SequencePN encoding 2A10 humanised heavy
chain H1 SEQ ID 79 83atgggatgga gctgtatcat cctcttcttg gtagcaacag
ctacaggtgt ccactcccag 60gtgcagctgg tgcagtctgg ggctgaggtg aagaagcctg
gggcctcagt gaaggtttcc 120tgcaaggcat ctggatacac cttcaccagc
tactggatgc actgggtgcg acaggcccct 180ggacaagggc ttgagtggat
gggaaatatt aatcctagca atggtggtac taactacaat 240gagaagttca
agagcagagt caccatgacc agggacacgt ccacgagcac agtctacatg
300gagctgagca gcctgagatc tgaggacacg gccgtgtatt actgtgaact
gggacagggc 360tactggggcc agggaacact agtcacagtc tcctcagcct
ccaccaaggg cccatcggtc 420ttccccctgg caccctcctc caagagcacc
tctgggggca cagcggccct gggctgcctg 480gtcaaggact acttccccga
accggtgacg gtgtcgtgga actcaggcgc cctgaccagc 540ggcgtgcaca
ccttcccggc tgtcctacag tcctcaggac tctactccct cagcagcgtg
600gtgaccgtgc cctccagcag cttgggcacc cagacctaca tctgcaacgt
gaatcacaag 660cccagcaaca ccaaggtgga caagaaagtt gagcccaaat
cttgtgacaa aactcacaca 720tgcccaccgt gcccagcacc tgaactcgcg
ggggcaccgt cagtcttcct cttcccccca 780aaacccaagg acaccctcat
gatctcccgg acccctgagg tcacatgcgt ggtggtggac 840gtgagccacg
aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat
900aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
ggtcagcgtc 960ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt
acaagtgcaa ggtctccaac 1020aaagccctcc cagcccccat cgagaaaacc
atctccaaag ccaaagggca gccccgagaa 1080ccacaggtgt acaccctgcc
cccatcccgg gatgagctga ccaagaacca ggtcagcctg 1140acctgcctgg
tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg
1200cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
ctccttcttc 1260ctctacagca agctcaccgt ggacaagagc aggtggcagc
aggggaacgt cttctcatgc 1320tccgtgatgc atgaggctct gcacaaccac
tacacgcaga agagcctctc cctgtctccg 1380ggtaaatga
138984717DNAArtificial SequencePN encoding 2A10 humanised light
chain construct L11 SEQ ID 90 84atgggatgga gctgtatcat cctcttcttg
gtagcaacag ctacaggtgt ccactccgat 60attgtgataa cccagtctcc actctccctg
cccgtcaccc ttggacagcc ggcctccatc 120tcctgcaggt ctagtaagag
tctcctatat aaggatggga agacatactt gaattggttt 180cagcagaggc
caggccaatc tccacagctc ctaatttatt tgatgtccac ccgtgcatct
240ggggtcccag acagattcag cggcggtggg tcaggcactg atttcacact
gaaaatcagc 300agggtggagg ctgaggatgt tggggtttat tactgccaac
aacttgtaga gtatccgctc 360acgtttggcc aggggaccaa gctggagatc
aaacgtacgg tggctgcacc atctgtcttc 420atcttcccgc catctgatga
gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 480aataacttct
atcccagaga ggccaaagta cagtggaagg tggacaacgc cctccaatcg
540ggtaactccc aggagagtgt cacagagcag gacagcaagg acagcaccta
cagcctcagc 600agcaccctga cgctgagcaa agcagactac gagaaacaca
aagtctacgc ctgcgaagtc 660acccatcagg gcctgagctc gcccgtcaca
aagagcttca acaggggaga gtgttag 71785113PRTArtificial Sequence2A10 VH
humanised construct H19 85Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala1 5 10 15Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp
Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40
45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60Lys Ser Arg Ala Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Ala
Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser 100 105 110Ser86113PRTArtificial Sequence2A10 VH
humanised construct H20 86Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser
Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met His Trp Val Arg Gln Ala
Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn
Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Met
Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser
Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105
110Ser87113PRTArtificial Sequence2A10 VH humanised construct H21
87Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser
Tyr 20 25 30Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn
Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Met Thr Arg Asp Thr Ser Thr
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln
Gly Thr Leu Val Thr Val Ser 100 105 110Ser88113PRTArtificial
Sequence2A10 VH humanised construct H22 88Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met His Trp
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser
Lys Ala Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
Ser 100 105 110Ser 89113PRTArtificial Sequence2A10 VH humanised
construct H23 89Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Ser Tyr 20 25 30Trp Met His Trp Val Lys Gln Arg Pro Gly Gln
Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr
Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Met Thr Val Asp
Thr Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr
Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105
110Ser90113PRTArtificial Sequence2A10 VH humanised construct H24
90Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser
Tyr 20 25 30Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn
Glu Lys Phe 50 55 60Lys Ser Lys Ala Thr Leu Thr Val Asp Thr Ser Thr
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln
Gly Thr Leu Val Thr Val Ser 100 105 110Ser91113PRTArtificial
Sequence2A10 VH humanised construct H25 91Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met His Trp
Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60Lys Ser
Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
Ser 100 105 110Ser92462PRTArtificial Sequence2A10 heavy chain
humanised construct H19 92Met Gly Trp Ser Cys Ile Ile Leu Phe Leu
Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp
Val Arg Gln Arg Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile
Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys
Ser Arg Ala Thr Met Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Ala Tyr
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr
Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120
125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His Thr Phe
Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser
Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230 235
240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe
245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360
365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 46093462PRTArtificial
Sequence2A10 heavy chain humanised construct H20 93Met Gly Trp Ser
Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly
Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr
Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu 50 55
60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65
70 75 80Glu Lys Phe Lys Ser Lys Ala Thr Met Thr Arg Asp Thr Ser Thr
Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr
Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln
Gly Thr Leu Val 115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
Ser Val Phe Pro Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly
Gly Thr Ala Ala Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200
205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
210 215 220Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
His Thr225 230 235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly
Ala Pro Ser Val Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu Met Ile Ser Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val
Asp Val Ser His Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr
Val Asp Gly Val Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg
Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315
320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile
Ala Val Glu Trp Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440
445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46094462PRTArtificial Sequence2A10 heavy chain humanised construct
H21 94Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Arg Gln Arg Pro
Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Lys Ala Thr Met
Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46095462PRTArtificial Sequence2A10 heavy
chain humanised construct H22 95Met Gly Trp Ser Cys Ile Ile Leu Phe
Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val
Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His
Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn
Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe
Lys Ser Lys Ala Thr Met Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Ala
Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105
110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225
230 235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser
Val Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
Ser Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345
350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46096462PRTArtificial Sequence2A10 heavy chain humanised construct
H23 96Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Lys Gln Arg Pro
Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Lys Ala Thr Met
Thr Val Asp Thr Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46097462PRTArtificial Sequence2A10 heavy
chain humanised construct H24 97Met Gly Trp Ser Cys Ile Ile Leu Phe
Leu Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val
Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His
Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60Glu Trp Ile Gly Asn
Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe
Lys Ser Lys Ala Thr Leu Thr Val Asp Thr Ser Thr Ser 85 90 95Thr Ala
Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105
110Tyr Tyr Cys Glu Leu Gly Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala 130 135 140Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu145 150 155 160Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly 165 170 175Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser 180 185 190Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195 200 205Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 210 215 220Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr225 230
235 240Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val
Phe 245 250 255Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro 260 265 270Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val 275 280 285Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr 290 295 300Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val305 310 315 320Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325 330 335Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 340 345
350Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
355 360 365Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
Leu Val 370 375 380Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser Asn Gly385 390 395 400Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser Asp 405 410 415Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp Lys Ser Arg Trp 420 425 430Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala Leu His 435 440 445Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455
46098462PRTArtificial Sequence2A10 heavy chain humanised construct
H25 98Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr
Gly1 5 10 15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys 20 25 30Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe 35 40 45Thr Ser Tyr Trp Met His Trp Val Arg Gln Arg Pro
Gly Gln Gly Leu 50 55 60Glu Trp Met Gly Asn Ile Asn Pro Ser Asn Gly
Gly Thr Asn Tyr Asn65 70 75 80Glu Lys Phe Lys Ser Arg Val Thr Met
Thr Arg Asp Thr Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Glu Leu Gly
Gln Gly Tyr Trp Gly Gln Gly Thr Leu Val 115 120 125Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu145 150 155
160Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser 180 185 190Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu 195 200 205Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr 210 215 220Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp Lys Thr His Thr225 230 235 240Cys Pro Pro Cys Pro
Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe 245 250 255Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260 265 270Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 275 280
285Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
290 295 300Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val305 310 315 320Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys 325 330 335Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345 350Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro 355 360 365Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370 375 380Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly385 390 395
400Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
405 410 415Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp 420 425 430Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His 435 440 445Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly Lys 450 455 46099339DNAArtificial SequencePN encoding
2A10 VH humanised construct H19 SEQ ID 85 99caggtgcagc tggtgcagtc
tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata
caccttcacc agctactgga tgcactgggt gcgacagagg 120cctggacaag
ggcttgagtg gatcggaaat attaatccta gcaatggtgg tactaactac
180aatgagaagt tcaagagcag agccaccatg accagggaca cgtccacgag
cacagcctac 240atggagctga gcagcctgag atctgaggac acggccgtgt
attactgtga actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
339100339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H20 SEQ ID 86 100caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gcgacaggcc 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa ggccaccatg accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
339101339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H21 SEQ ID 87 101caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gcgacagagg 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa ggccaccatg accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
339102339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H22 SEQ ID 88 102caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaagcagagg 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa ggccaccatg accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
339103339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H23 SEQ ID 89 103caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaagcagagg 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa ggccaccatg accgtagaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
339104339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H24 SEQ ID 90 104caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gaagcagagg 120cctggacaag ggcttgagtg
gatcggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcaa ggccaccctg accgtagaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
339105339DNAArtificial SequencePN encoding 2A10 VH humanised
construct H25 SEQ ID 91 105caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc
agctactgga tgcactgggt gcgacagagg 120cctggacaag ggcttgagtg
gatgggaaat attaatccta gcaatggtgg tactaactac 180aatgagaagt
tcaagagcag agtcaccatg accagggaca cgtccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtga
actgggacag 300ggctactggg gccagggaac actagtcaca gtctcctca
3391061389DNAArtificial SequencePN encoding 2A10 heavy chain
humanised construct H19 SEQ ID 92 106atgggatgga gctgtatcat
cctcttcttg gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg tgcagtctgg
ggctgaggtg aagaagcctg gggcctcagt gaaggtttcc 120tgcaaggcat
ctggatacac cttcaccagc tactggatgc actgggtgcg acagaggcct
180ggacaagggc ttgagtggat cggaaatatt aatcctagca atggtggtac
taactacaat 240gagaagttca agagcagagc caccatgacc agggacacgt
ccacgagcac agcctacatg 300gagctgagca gcctgagatc tgaggacacg
gccgtgtatt actgtgaact gggacagggc 360tactggggcc agggaacact
agtcacagtc tcctcagcct ccaccaaggg cccatcggtc 420ttccccctgg
caccctcctc caagagcacc tctgggggca cagcggccct gggctgcctg
480gtcaaggact acttccccga accggtgacg gtgtcgtgga actcaggcgc
cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag tcctcaggac
tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag cttgggcacc
cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca ccaaggtgga
caagaaagtt gagcccaaat cttgtgacaa aactcacaca 720tgcccaccgt
gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct cttcccccca
780aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc aactggtacg
tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg ggaggagcag
tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc tgcaccagga
ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac 1020aaagccctcc
cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa
1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga ccaagaacca
ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc gacatcgccg
tggagtggga gagcaatggg 1200cagccggaga acaactacaa gaccacgcct
cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca agctcaccgt
ggacaagagc aggtggcagc aggggaacgt cttctcatgc 1320tccgtgatgc
atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg
1380ggtaaatga 13891071389DNAArtificial SequencePN encoding 2A10
heavy chain humanised construct H20 SEQ ID 93 107atgggatgga
gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg
tgcagtctgg ggctgaggtg aagaagcctg gggcctcagt gaaggtttcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgcg
acaggcccct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcaaggc caccatgacc
agggacacgt ccacgagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacacg gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct
ccaccaaggg cccatcggtc 420ttccccctgg caccctcctc caagagcacc
tctgggggca cagcggccct gggctgcctg 480gtcaaggact acttccccga
accggtgacg gtgtcgtgga actcaggcgc cctgaccagc 540ggcgtgcaca
ccttcccggc tgtcctacag tcctcaggac tctactccct cagcagcgtg
600gtgaccgtgc cctccagcag cttgggcacc cagacctaca tctgcaacgt
gaatcacaag 660cccagcaaca ccaaggtgga caagaaagtt gagcccaaat
cttgtgacaa aactcacaca 720tgcccaccgt gcccagcacc tgaactcgcg
ggggcaccgt cagtcttcct cttcccccca 780aaacccaagg acaccctcat
gatctcccgg acccctgagg tcacatgcgt ggtggtggac 840gtgagccacg
aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat
900aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
ggtcagcgtc 960ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt
acaagtgcaa ggtctccaac 1020aaagccctcc cagcccccat cgagaaaacc
atctccaaag ccaaagggca gccccgagaa 1080ccacaggtgt acaccctgcc
cccatcccgg gatgagctga ccaagaacca ggtcagcctg 1140acctgcctgg
tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg
1200cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
ctccttcttc 1260ctctacagca agctcaccgt ggacaagagc aggtggcagc
aggggaacgt cttctcatgc 1320tccgtgatgc atgaggctct gcacaaccac
tacacgcaga agagcctctc cctgtctccg 1380ggtaaatga
13891081389DNAArtificial SequencePN encoding 2A10 heavy chain
humanised construct H21 SEQ ID 94 108atgggatgga gctgtatcat
cctcttcttg gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg tgcagtctgg
ggctgaggtg aagaagcctg gggcctcagt gaaggtttcc 120tgcaaggcat
ctggatacac cttcaccagc tactggatgc actgggtgcg acagaggcct
180ggacaagggc ttgagtggat cggaaatatt aatcctagca atggtggtac
taactacaat 240gagaagttca agagcaaggc caccatgacc agggacacgt
ccacgagcac agcctacatg 300gagctgagca gcctgagatc tgaggacacg
gccgtgtatt actgtgaact gggacagggc 360tactggggcc agggaacact
agtcacagtc tcctcagcct ccaccaaggg cccatcggtc 420ttccccctgg
caccctcctc caagagcacc tctgggggca cagcggccct gggctgcctg
480gtcaaggact acttccccga accggtgacg gtgtcgtgga actcaggcgc
cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag tcctcaggac
tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag cttgggcacc
cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca ccaaggtgga
caagaaagtt gagcccaaat cttgtgacaa aactcacaca 720tgcccaccgt
gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct cttcccccca
780aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc aactggtacg
tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg ggaggagcag
tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc tgcaccagga
ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac 1020aaagccctcc
cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa
1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga ccaagaacca
ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc gacatcgccg
tggagtggga gagcaatggg 1200cagccggaga acaactacaa gaccacgcct
cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca agctcaccgt
ggacaagagc aggtggcagc aggggaacgt cttctcatgc 1320tccgtgatgc
atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg
1380ggtaaatga 13891091389DNAArtificial SequencePN encoding 2A10
heavy chain humanised construct H22 SEQ ID 95 109atgggatgga
gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactcccag 60gtgcagctgg
tgcagtctgg ggctgaggtg aagaagcctg gggcctcagt gaaggtttcc
120tgcaaggcat ctggatacac cttcaccagc tactggatgc actgggtgaa
gcagaggcct 180ggacaagggc ttgagtggat cggaaatatt aatcctagca
atggtggtac taactacaat 240gagaagttca agagcaaggc caccatgacc
agggacacgt ccacgagcac agcctacatg 300gagctgagca gcctgagatc
tgaggacacg gccgtgtatt actgtgaact gggacagggc 360tactggggcc
agggaacact agtcacagtc tcctcagcct ccaccaaggg cccatcggtc
420ttccccctgg caccctcctc caagagcacc tctgggggca cagcggccct
gggctgcctg 480gtcaaggact acttccccga accggtgacg gtgtcgtgga
actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc tgtcctacag
tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc cctccagcag
cttgggcacc cagacctaca tctgcaacgt gaatcacaag 660cccagcaaca
ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactcacaca
720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt cagtcttcct
cttcccccca 780aaacccaagg acaccctcat gatctcccgg acccctgagg
tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga ggtcaagttc
aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga caaagccgcg
ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960ctcaccgtcc
tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac
1020aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg gatgagctga
ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt ctatcccagc
gacatcgccg tggagtggga gagcaatggg 1200cagccggaga acaactacaa
gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260ctctacagca
agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc
1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
cctgtctccg 1380ggtaaatga 13891101389DNAArtificial SequencePN
encoding 2A10 heavy chain humanised construct H23 SEQ ID 96
110atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt
ccactcccag 60gtgcagctgg tgcagtctgg ggctgaggtg aagaagcctg gggcctcagt
gaaggtttcc 120tgcaaggcat ctggatacac cttcaccagc tactggatgc
actgggtgaa gcagaggcct 180ggacaagggc ttgagtggat cggaaatatt
aatcctagca atggtggtac taactacaat 240gagaagttca agagcaaggc
caccatgacc gtagacacgt ccacgagcac agcctacatg 300gagctgagca
gcctgagatc tgaggacacg gccgtgtatt actgtgaact gggacagggc
360tactggggcc agggaacact agtcacagtc tcctcagcct ccaccaaggg
cccatcggtc 420ttccccctgg caccctcctc caagagcacc tctgggggca
cagcggccct gggctgcctg 480gtcaaggact acttccccga accggtgacg
gtgtcgtgga actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc
tgtcctacag tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc
cctccagcag cttgggcacc cagacctaca tctgcaacgt gaatcacaag
660cccagcaaca ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa
aactcacaca 720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt
cagtcttcct cttcccccca 780aaacccaagg acaccctcat gatctcccgg
acccctgagg tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga
ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga
caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc
960ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
ggtctccaac 1020aaagccctcc cagcccccat cgagaaaacc atctccaaag
ccaaagggca gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg
gatgagctga ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt
ctatcccagc gacatcgccg tggagtggga gagcaatggg 1200cagccggaga
acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc
1260ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
cttctcatgc 1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga
agagcctctc cctgtctccg 1380ggtaaatga 13891111389DNAArtificial
SequencePN encoding 2A10 heavy chain humanised construct H24 SEQ ID
97 111atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt
ccactcccag 60gtgcagctgg tgcagtctgg ggctgaggtg aagaagcctg gggcctcagt
gaaggtttcc 120tgcaaggcat ctggatacac cttcaccagc tactggatgc
actgggtgaa gcagaggcct 180ggacaagggc ttgagtggat cggaaatatt
aatcctagca atggtggtac taactacaat 240gagaagttca agagcaaggc
caccctgacc gtagacacgt ccacgagcac agcctacatg 300gagctgagca
gcctgagatc tgaggacacg gccgtgtatt actgtgaact gggacagggc
360tactggggcc agggaacact agtcacagtc tcctcagcct ccaccaaggg
cccatcggtc 420ttccccctgg caccctcctc caagagcacc tctgggggca
cagcggccct gggctgcctg 480gtcaaggact acttccccga accggtgacg
gtgtcgtgga actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc
tgtcctacag tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc
cctccagcag cttgggcacc cagacctaca tctgcaacgt gaatcacaag
660cccagcaaca ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa
aactcacaca 720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt
cagtcttcct cttcccccca 780aaacccaagg acaccctcat gatctcccgg
acccctgagg tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga
ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga
caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc
960ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
ggtctccaac 1020aaagccctcc cagcccccat cgagaaaacc atctccaaag
ccaaagggca gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg
gatgagctga ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt
ctatcccagc gacatcgccg tggagtggga gagcaatggg 1200cagccggaga
acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc
1260ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
cttctcatgc 1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga
agagcctctc cctgtctccg 1380ggtaaatga 13891121389DNAArtificial
SequencePN encoding 2A10 heavy chain humanised construct H25 SEQ ID
98 112atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt
ccactcccag 60gtgcagctgg tgcagtctgg ggctgaggtg aagaagcctg gggcctcagt
gaaggtttcc 120tgcaaggcat ctggatacac cttcaccagc tactggatgc
actgggtgcg acagaggcct 180ggacaagggc ttgagtggat gggaaatatt
aatcctagca atggtggtac taactacaat 240gagaagttca agagcagagt
caccatgacc agggacacgt ccacgagcac agcctacatg 300gagctgagca
gcctgagatc tgaggacacg gccgtgtatt actgtgaact gggacagggc
360tactggggcc agggaacact agtcacagtc tcctcagcct ccaccaaggg
cccatcggtc 420ttccccctgg caccctcctc caagagcacc tctgggggca
cagcggccct gggctgcctg 480gtcaaggact acttccccga accggtgacg
gtgtcgtgga actcaggcgc cctgaccagc 540ggcgtgcaca ccttcccggc
tgtcctacag tcctcaggac tctactccct cagcagcgtg 600gtgaccgtgc
cctccagcag cttgggcacc cagacctaca tctgcaacgt gaatcacaag
660cccagcaaca ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa
aactcacaca 720tgcccaccgt gcccagcacc tgaactcgcg ggggcaccgt
cagtcttcct cttcccccca 780aaacccaagg acaccctcat gatctcccgg
acccctgagg tcacatgcgt ggtggtggac 840gtgagccacg aagaccctga
ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 900aatgccaaga
caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc
960ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
ggtctccaac 1020aaagccctcc cagcccccat cgagaaaacc atctccaaag
ccaaagggca gccccgagaa 1080ccacaggtgt acaccctgcc cccatcccgg
gatgagctga ccaagaacca ggtcagcctg 1140acctgcctgg tcaaaggctt
ctatcccagc gacatcgccg tggagtggga gagcaatggg 1200cagccggaga
acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc
1260ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
cttctcatgc 1320tccgtgatgc atgaggctct gcacaaccac tacacgcaga
agagcctctc cctgtctccg 1380ggtaaatga 1389113200PRTArtificial
SequenceAmino acid sequence of Marmoset NOGO-A fragment 113Val Gln
Asp Ser Leu Cys Pro Val Ala Gln Leu Cys Pro Ser Phe Glu1 5 10 15Glu
Ser Glu Ala Thr Pro Ser Pro Val Leu Pro Asp Ile Val Met Glu 20 25
30Ala Pro Leu Asn Ser Ala Val Pro Ser Ala Gly Ala Ser Ala Val Gln
35 40 45Pro Ser Ser Ser Pro Leu Glu Ala Ser Ser Val Asn Phe Glu Ser
Val 50 55 60Lys His Glu Pro Glu Asn Pro Pro Pro Tyr Glu Glu Ala Met
Asn Val65 70 75 80Ser Arg Lys Lys Val Ser Gly Ile Lys Glu Glu Ile
Lys Glu Pro Glu 85 90 95Ser Ile Asn Ala Ala Val Gln Glu Thr Glu Ala
Pro Tyr Ile Ser Ile 100 105 110Ala Cys Asp Leu Ile Lys Glu Thr Lys
Leu Ser Ala Glu Pro Thr Pro 115 120 125Asp Phe Ser Ser Tyr Ser Glu
Met Ala Lys Val Glu Gln Pro Leu Pro 130 135 140Asp His Ser Glu Leu
Val Glu Asp Ser Ser Pro Asp Ser Glu Pro Val145 150 155 160Asp Leu
Phe Ser Asp Asp Ser Ile Pro Asp Val Pro Gln Lys Gln Asp 165 170
175Glu Ala Val Ile Leu Val Lys Glu Thr Leu Thr Glu Thr Ser Phe Glu
180 185 190Ser Met Ile Glu His Glu Asn Lys 195 200
* * * * *