Methods for preventing and treating heart disease

Abstract

The present disclosure relates to novel uses and methods for preventing and / or treating heart disease using a therapeutically effective amount of an ActRII receptor antagonist, e.g., an ActRII receptor binding molecule, e.g., an ActRII receptor antibody, such as bimagrumab antibody.

Description

[0001] This application claims priority to U.S. Provisional Application No. 62 / 476,054, filed on March 24, 2017. Technical Field

[0002] The present disclosure is in the field of type II activin receptor (ActRII) antagonists, such as molecules that can antagonize the binding of activin, growth differentiation factor (GDF), bone morphogenetic protein (BMP) and myostatin to the human Actll receptor, such as antagonist antibodies to ActRIIA and / or ActRIIB, such as bimagrumab. Specifically, the present disclosure relates to the prevention and / or treatment of heart failure, including heart failure with reduced ejection fraction (HFrEF) and heart failure with preserved ejection fraction (HFpEF), and to treating structural and / or functional cardiac abnormalities associated with the condition, such as valvular heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and hereditary or idiopathic dilated cardiomyopathy, by administering a therapeutically effective amount of an ActRII receptor antagonist to a subject. Notably, these conditions include clinical syndromes that often coexist but can occur alone and are sometimes referred to as systolic and / or diastolic heart failure, left-sided and / or right-sided heart failure, and congestive heart failure. Background Art

[0003] Activin receptor type IIB (ActRIIB) is a signaling receptor for various members of the transforming growth factor beta (TGF-β) superfamily. Members of this family include activin A, nodal, BMP2, BMP6, BMP7, BMP9, GDF5, GDF8 (myostatin), and GDF11, all of which are involved in the negative regulation of muscle (Akpan et al., 2009).

[0004] Myostatin (GDF8) acts via type II activin receptors (primarily via ActRIIB), and its proposed signaling is via the SMAD 2 / 3 pathway, which is involved in the inhibition of protein synthesis, as well as myocyte differentiation and proliferation. Myostatin inhibition or genetic ablation increases muscle mass and strength (Lee et al. 2005, Lee and McPherron 2001, Whittemore et al. 2003).

[0005] Bimagrumab, also known as BYM338, is a monoclonal antibody developed to competitively bind to the type IIB activin receptor (ActRII) with an affinity greater than that of the natural ligands of the type IIB activin receptor, myostatin or activin. Bimagrumab is disclosed in WO2010 / 125003, which is incorporated herein by reference as if fully set forth. Bimagrumab is a fully human antibody (modified IgG1, 234-235-Ala-Ala, λ2) that binds to the ligand binding domains of ActRIIA and ActRIIB, thereby preventing the binding and subsequent signaling of ligands of ActRIIA and ActRIIB, including myostatin and activin, which act as natural inhibitors of skeletal muscle growth. Bimagrumab cross-reacts with human and mouse ActRIIB and is effective in human, macaque, mouse and rat skeletal muscle cells. ActRIIB is widely distributed in skeletal muscle, adipose tissue, and various organs including the heart (Rebbapragada et al., 2003).

[0006] Heart failure is a clinical syndrome in which impaired cardiac function results in insufficient systemic perfusion to meet the body's metabolic demands. Heart failure is divided into two major categories: (1) heart failure with a reduced ejection fraction (HFrEF) (also known as "systolic heart failure") and (2) heart failure with a preserved ejection fraction (HFpEF) (also known as "diastolic heart failure"). In HFrEF, reduced myocardial contractility is the primary mechanism that impairs cardiac output and leads to insufficient systemic perfusion. In HFpEF, resting myocardial contractility is generally preserved. However, a variety of other defects in cardiac function, including cardiac reserve and diastolic function, impair the functional performance of the heart, resulting in a similar phenotype to clinical heart failure. Various conditions can damage or weaken the heart and lead to heart failure, including, for example, valvular heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and genetic and / or idiopathic dilated cardiomyopathy.

[0007] Heart failure caused by causes such as diabetes, aging, hypertension, ischemic heart disease, coronary heart disease, valvular heart disease, and hereditary and idiopathic cardiomyopathies is a major cause of morbidity and mortality worldwide. For this increasingly common disease process, available drug therapies are limited. The standard of care for heart failure includes a variety of drug therapies targeting the various mechanisms involved in the complex pathophysiology of the disease. Unfortunately, even with guideline-directed therapy, the prognosis for these patients remains poor, with a 5-year mortality rate approaching 50%. In advanced systolic heart failure, patients may not tolerate commonly used oral drug therapies due to hemodynamic, nephrogenic, and arrhythmogenic side effects, or may not be able to achieve sufficient relief from such therapies. For these patients, advanced therapies such as intravenous inotropes, mechanical support devices, and heart transplantation are very limited, expensive, and have significant risks.

[0008] Prior to the present disclosure, targeted inhibition of the type II activin receptor (ActRIIA / B) had not been investigated as a prophylactic or therapeutic approach for heart failure or the aforementioned conditions that can lead to heart failure. As disclosed herein, there is now evidence that systemic administration of an ActRIIA / B receptor antagonist, such as CDD866, which is a murinized version of BYM338 (in which the human Fc region of the antibody has been replaced by a mouse Fc), has significant beneficial effects on cardiac function in mice subjected to transverse aortic constriction (TAC). TAC is a commonly used experimental model of cardiac hypertrophy and heart failure caused by pressure overload. First validated by Rockman et al., 1991, the murine TAC model has been widely used since 1991 as a valuable tool for modeling human cardiovascular disease and understanding the fundamental signaling processes involved in the development of cardiac hypertrophic responses and heart failure (deAlmeida et al., 2010). As disclosed herein, CDD866 not only prevents TAC-mediated cardiac dysfunction but is also able to restore cardiac function within 1-2 weeks of drug administration after heart failure has established. It also enhances skeletal muscle growth (which tends to atrophy in advanced forms of heart failure) and induces minimal cardiac effects in control mice that have not been subjected to pathological stress / injury from TAC.

[0009] Disclosed herein are ActRII receptor antagonists for use in treating and / or preventing heart failure, including heart failure caused by or associated with conditions such as valvular heart disease, ischemic heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and hereditary and / or idiopathic dilated cardiomyopathy. Also disclosed are ActRII receptor antagonists for use in treating structural and / or functional cardiac abnormalities associated with the above-mentioned conditions. Also provided are methods of using such ActRII antagonists to treat and / or prevent heart failure, as well as treating structural and / or functional cardiac abnormalities associated with the above-mentioned conditions. Summary of the Invention

[0010] Disclosed herein are ActRII receptor antagonists for the treatment and / or prevention of heart failure. Heart failure may be caused by or associated with a variety of conditions, such as valvular disease (such as aortic stenosis), coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and hereditary or idiopathic dilated cardiomyopathy. Included herein are heart failure with reduced ejection fraction (HFrEF) and heart failure with preserved ejection fraction (HFpEF).

[0011] Also disclosed herein are ActRII receptor antagonists for the treatment of structural and / or functional cardiac abnormalities associated with conditions such as valvular heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and hereditary or idiopathic dilated cardiomyopathy. In some cases, peripartum cardiomyopathy occurs in late pregnancy or during the 6 months postpartum. Stress cardiomyopathy typically occurs in elderly women in the postmenopausal period. An example of valvular heart disease is aortic stenosis, which can be associated with frailty and / or sarcopenia. Stress cardiomyopathy can occur following psychological, pathological, or physical stress.

[0012] Disclosed herein are methods for treating and / or preventing heart failure. The methods include administering a therapeutically effective amount of an ActRII receptor antagonist, such as bimagrumab, to a subject having heart failure or at risk of developing heart failure. Included herein are heart failure with a reduced ejection fraction (HFrEF) and heart failure with a preserved ejection fraction (HFpEF). Heart failure can be diagnosed in a patient using well-known methods, including, for example, measuring brain natriuretic peptide, followed by cardiac ultrasound if the brain natriuretic peptide is positive, and performing imaging such as echocardiography.

[0013] Patients are at risk for developing heart failure when they have conditions such as valvular heart disease, coronary artery disease (including previous myocardial infarction), hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, and hereditary or idiopathic dilated cardiomyopathy.

[0014] Also disclosed herein are methods for treating structural and / or functional cardiac abnormalities associated with conditions such as valvular heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and hereditary or idiopathic dilated cardiomyopathy, comprising administering an effective amount of an ActRII receptor antagonist to a subject having such structural and / or functional cardiac abnormalities associated with such conditions.

[0015] Examples of ActRII receptor antagonists for use in the methods described herein are ActRII receptor binding molecules that block access of ActRII interacting ligands such as myostatin, GDF11, and activin A to ActRII. ActRII receptor binding molecules can bind to ActRIIA and / or to ActRIIB receptors. Examples of ActRII binding molecules include, but are not limited to, antibodies that bind to ActRIIA and / or ActRIIB receptors, such as anti-ActRII receptor antibodies. Preferably, the anti-ActRII receptor antibody is BYM338, also known as bimagrumab.

[0016] Another example of an ActRII receptor antagonist for use in the methods described herein is a soluble form of the extracellular domain of the ActRIIA receptor or the ActRIIB receptor that binds to an ActRII-interacting ligand, such as myostatin, GDF11, and activin A. This "receptor-body" inhibits the function of a cell-bound ActRII receptor by competing away its ligand.

[0017] Disclosed herein are ActRII receptor antagonists for use in or for use in the methods described herein, wherein the ActRII receptor antagonist is an anti-ActRII antibody that binds to an epitope of ActRIIB consisting of amino acids 19-134 of SEQ ID NO: 181 (SEQ ID NO: 182).

[0018] Disclosed herein are ActRII receptor antagonists for use in or in the methods described herein, wherein the anti-ActRII antibody binds to an epitope of ActRIIB comprising or consisting of:

[0019] (a) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188);

[0020] (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC-SEQ ID NO: 186);

[0021] (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190);

[0022] (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR—SEQ ID NO: 189);

[0023] (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW-SEQ ID NO: 187);

[0024] (f) amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191);

[0025] (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN - SEQ ID NO: 192); or

[0026] (h) Amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR).

[0027] Other anti-ActRIIB antibodies for use in the methods described herein include, for example

[0028] a) an anti-ActRIIB antibody that binds to an epitope of ActRIIB, the epitope comprising:

[0029] (a) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188);

[0030] (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC-SEQ ID NO: 186);

[0031] (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190);

[0032] (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR—SEQ ID NO: 189);

[0033] (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW-SEQ ID NO: 187);

[0034] (f) amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191);

[0035] (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN - SEQ ID NO: 192); or

[0036] (h) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR); and

[0037] b) an ActRIIB antagonist antibody that binds to an epitope of ActRIIB comprising amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188);

[0038] (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC-SEQ ID NO: 186);

[0039] (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190);

[0040] (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR—SEQ ID NO: 189);

[0041] (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW-SEQ ID NO: 187);

[0042] (f) amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191);

[0043] (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN - SEQ ID NO: 192); or

[0044] (h) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR), wherein the K D It is about 2pM.

[0045] In one embodiment, an ActRII receptor antagonist for use in the methods described herein is an antibody that binds to ActRIIB with an affinity that is about 10 times greater than that of the antibody that binds to ActRIIA.

[0046] The ActRII receptor antagonist used in or for use in the methods described herein can be an antibody comprising: a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14; a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28; a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: 42; a light chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 43 to SEQ ID NO: 56; a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 57 to SEQ ID NO: 70; and a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 71 to SEQ ID NO: 84.

[0047] An ActRII receptor antagonist for use in the methods described herein can be an antibody comprising:

[0048] (a) the heavy chain variable region CDR1 set forth in SEQ ID NO: 1; the heavy chain variable region CDR2 set forth in SEQ ID NO: 15; the heavy chain variable region CDR3 set forth in SEQ ID NO: 29; the light chain variable region CDR1 set forth in SEQ ID NO: 43; the light chain variable region CDR2 set forth in SEQ ID NO: 57; and the light chain variable region CDR3 set forth in SEQ ID NO: 71,

[0049] (b) the heavy chain variable region CDR1 set forth in SEQ ID NO: 2; the heavy chain variable region CDR2 set forth in SEQ ID NO: 16; the heavy chain variable region CDR3 set forth in SEQ ID NO: 30; the light chain variable region CDR1 set forth in SEQ ID NO: 44; the light chain variable region CDR2 set forth in SEQ ID NO: 58; and the light chain variable region CDR3 set forth in SEQ ID NO: 72,

[0050] (c) the heavy chain variable region CDR1 set forth in SEQ ID NO: 3; the heavy chain variable region CDR2 set forth in SEQ ID NO: 17; the heavy chain variable region CDR3 set forth in SEQ ID NO: 31; the light chain variable region CDR1 set forth in SEQ ID NO: 45; the light chain variable region CDR2 set forth in SEQ ID NO: 59; and the light chain variable region CDR3 set forth in SEQ ID NO: 73,

[0051] (d) the heavy chain variable region CDR1 set forth in SEQ ID NO:4; the heavy chain variable region CDR2 set forth in SEQ ID NO:18; the heavy chain variable region CDR3 set forth in SEQ ID NO:32; the light chain variable region CDR1 set forth in SEQ ID NO:46; the light chain variable region CDR2 set forth in SEQ ID NO:60; and the light chain variable region CDR3 set forth in SEQ ID NO:74,

[0052] (e) the heavy chain variable region CDR1 set forth in SEQ ID NO:5; the heavy chain variable region CDR2 set forth in SEQ ID NO:19; the heavy chain variable region CDR3 set forth in SEQ ID NO:33; the light chain variable region CDR1 set forth in SEQ ID NO:47; the light chain variable region CDR2 set forth in SEQ ID NO:61; and the light chain variable region CDR3 set forth in SEQ ID NO:75,

[0053] (f) the heavy chain variable region CDR1 set forth in SEQ ID NO:6; the heavy chain variable region CDR2 set forth in SEQ ID NO:20; the heavy chain variable region CDR3 set forth in SEQ ID NO:34; the light chain variable region CDR1 set forth in SEQ ID NO:48; the light chain variable region CDR2 set forth in SEQ ID NO:62; and the light chain variable region CDR3 set forth in SEQ ID NO:76,

[0054] (g) the heavy chain variable region CDR1 set forth in SEQ ID NO:7; the heavy chain variable region CDR2 set forth in SEQ ID NO:21; the heavy chain variable region CDR3 set forth in SEQ ID NO:35; the light chain variable region CDR1 set forth in SEQ ID NO:49; the light chain variable region CDR2 set forth in SEQ ID NO:63; and the light chain variable region CDR3 set forth in SEQ ID NO:77,

[0055] (h) the heavy chain variable region CDR1 set forth in SEQ ID NO:8; the heavy chain variable region CDR2 set forth in SEQ ID NO:22; the heavy chain variable region CDR3 set forth in SEQ ID NO:36; the light chain variable region CDR1 set forth in SEQ ID NO:50; the light chain variable region CDR2 set forth in SEQ ID NO:64; and the light chain variable region CDR3 set forth in SEQ ID NO:78,

[0056] (i) the heavy chain variable region CDR1 set forth in SEQ ID NO:9; the heavy chain variable region CDR2 set forth in SEQ ID NO:23; the heavy chain variable region CDR3 set forth in SEQ ID NO:37; the light chain variable region CDR1 set forth in SEQ ID NO:51; the light chain variable region CDR2 set forth in SEQ ID NO:65; and the light chain variable region CDR3 set forth in SEQ ID NO:79,

[0057] (j) the heavy chain variable region CDR1 set forth in SEQ ID NO:10; the heavy chain variable region CDR2 set forth in SEQ ID NO:24; the heavy chain variable region CDR3 set forth in SEQ ID NO:38; the light chain variable region CDR1 set forth in SEQ ID NO:52; the light chain variable region CDR2 set forth in SEQ ID NO:66; and the light chain variable region CDR3 set forth in SEQ ID NO:80,

[0058] (k) the heavy chain variable region CDR1 set forth in SEQ ID NO:11; the heavy chain variable region CDR2 set forth in SEQ ID NO:25; the heavy chain variable region CDR3 set forth in SEQ ID NO:39; the light chain variable region CDR1 set forth in SEQ ID NO:53; the light chain variable region CDR2 set forth in SEQ ID NO:67; and the light chain variable region CDR3 set forth in SEQ ID NO:81,

[0059] (1) the heavy chain variable region CDR1 set forth in SEQ ID NO:12; the heavy chain variable region CDR2 set forth in SEQ ID NO:26; the heavy chain variable region CDR3 set forth in SEQ ID NO:40; the light chain variable region CDR1 set forth in SEQ ID NO:54; the light chain variable region CDR2 set forth in SEQ ID NO:68; and the light chain variable region CDR3 set forth in SEQ ID NO:82,

[0060] (m) the heavy chain variable region CDR1 set forth in SEQ ID NO:13; the heavy chain variable region CDR2 set forth in SEQ ID NO:27; the heavy chain variable region CDR3 set forth in SEQ ID NO:41; the light chain variable region CDR1 set forth in SEQ ID NO:55; the light chain variable region CDR2 set forth in SEQ ID NO:69; and the light chain variable region CDR3 set forth in SEQ ID NO:83, or

[0061] (n) the heavy chain variable region CDR1 shown in SEQ ID NO:14; the heavy chain variable region CDR2 shown in SEQ ID NO:28; the heavy chain variable region CDR3 shown in SEQ ID NO:42; the light chain variable region CDR1 shown in SEQ ID NO:56; the light chain variable region CDR2 shown in SEQ ID NO:70; and the light chain variable region CDR3 shown in SEQ ID NO:84.

[0062] In another embodiment, the ActRII receptor antagonist for use in the methods described herein can be an antibody comprising a full-length heavy chain amino acid sequence having at least 95% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160.

[0063] The ActRII receptor antagonist for use in the methods described herein can be an antibody comprising a full-length light chain amino acid sequence having at least 95% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: 155.

[0064] An ActRII receptor antagonist for use in the methods described herein can be an antibody comprising:

[0065] (a) the variable heavy chain sequence shown in SEQ ID NO: 99 and the variable light chain sequence shown in SEQ ID NO: 85;

[0066] (b) the variable heavy chain sequence shown in SEQ ID NO: 100 and the variable light chain sequence shown in SEQ ID NO: 86;

[0067] (c) the variable heavy chain sequence shown in SEQ ID NO: 101 and the variable light chain sequence shown in SEQ ID NO: 87;

[0068] (d) the variable heavy chain sequence shown in SEQ ID NO: 102 and the variable light chain sequence shown in SEQ ID NO: 88;

[0069] (e) the variable heavy chain sequence shown in SEQ ID NO: 103 and the variable light chain sequence shown in SEQ ID NO: 89;

[0070] (f) the variable heavy chain sequence shown in SEQ ID NO: 104 and the variable light chain sequence shown in SEQ ID NO: 90;

[0071] (g) the variable heavy chain sequence shown in SEQ ID NO: 105 and the variable light chain sequence shown in SEQ ID NO: 91;

[0072] (h) the variable heavy chain sequence shown in SEQ ID NO: 106 and the variable light chain sequence shown in SEQ ID NO: 92;

[0073] (i) the variable heavy chain sequence set forth in SEQ ID NO: 107 and the variable light chain sequence set forth in SEQ ID NO: 93;

[0074] (j) the variable heavy chain sequence shown in SEQ ID NO: 108 and the variable light chain sequence shown in SEQ ID NO: 94;

[0075] (k) the variable heavy chain sequence shown in SEQ ID NO: 109 and the variable light chain sequence shown in SEQ ID NO: 95;

[0076] (1) the variable heavy chain sequence shown in SEQ ID NO: 110 and the variable light chain sequence shown in SEQ ID NO: 96;

[0077] (m) the variable heavy chain sequence shown in SEQ ID NO: 111 and the variable light chain sequence shown in SEQ ID NO: 97; or

[0078] (n) the variable heavy chain sequence shown in SEQ ID NO: 112 and the variable light chain sequence shown in SEQ ID NO: 98.

[0079] An ActRII receptor antagonist for use in the methods described herein can be an antibody comprising:

[0080] (a) the heavy chain sequence shown in SEQ ID NO: 146 and the light chain sequence shown in SEQ ID NO: 141;

[0081] (b) the heavy chain sequence shown in SEQ ID NO: 147 and the light chain sequence shown in SEQ ID NO: 142;

[0082] (c) the heavy chain sequence shown in SEQ ID NO: 148 and the light chain sequence shown in SEQ ID NO: 143;

[0083] (d) the heavy chain sequence shown in SEQ ID NO: 149 and the light chain sequence shown in SEQ ID NO: 144;

[0084] (e) the heavy chain sequence shown in SEQ ID NO: 150 and the light chain sequence shown in SEQ ID NO: 145;

[0085] (f) the heavy chain sequence shown in SEQ ID NO: 156 and the light chain sequence shown in SEQ ID NO: 151;

[0086] (g) the heavy chain sequence shown in SEQ ID NO: 157 and the light chain sequence shown in SEQ ID NO: 152;

[0087] (h) the heavy chain sequence shown in SEQ ID NO: 158 and the light chain sequence shown in SEQ ID NO: 153;

[0088] (i) the heavy chain sequence shown in SEQ ID NO: 159 and the light chain sequence shown in SEQ ID NO: 154; or

[0089] (j) The heavy chain sequence shown in SEQ ID NO: 160 and the light chain sequence shown in SEQ ID NO: 155.

[0090] Also disclosed are ActRII receptor antagonists for use in or with the methods described herein, wherein the ActRII receptor antagonist is an anti-ActRII receptor antibody that cross-blocks or is cross-blocked by at least one of the antibodies described above.

[0091] The ActRII receptor antagonist used in or for use in the methods described herein can be an anti-ActRII receptor antibody having altered effector function by mutation of the Fc region.

[0092] Exemplary of an antibody for use in or with the methods described herein is an anti-ActRII antibody encoded by pBW522 (DSM22873) or pBW524 (DSM22874).

[0093] The working examples described herein utilize CDD866, which is a murinized version of BYM338 in which the human Fc region of the antibody has been replaced with a mouse Fc.

[0094] However, a preferred antibody for use in or with the methods described herein is bimagrumab (BYM338), which is a fully human antibody (modified IgG1, 234-235-Ala-Ala, λ2).

[0095] "ActRII binding molecule" refers to any molecule that is capable of binding to a human ActRII receptor (ActRIIA and / or ActRIIB), either alone or in association with other molecules. The binding reaction can be shown by standard methods (qualitative assays), including, for example, binding assays with reference to negative control tests, competition assays, or bioassays for determining inhibition of binding of the ActRII receptor to myostatin, or any type of binding assay in which an antibody with an irrelevant specificity but ideally the same isotype, such as an anti-CD25 antibody, is used. Non-limiting examples of ActRII receptor binding molecules include small molecules, such as aptamers or other nucleic acid molecules designed to and / or undergo binding to the receptor, ligand decoys, and antibodies to the ActRII receptor produced by B cells, or hybridomas and chimeric CDR-grafted or human antibodies or any fragment thereof (e.g., F(ab')2 fragments and Fab fragments), as well as single chain or single domain antibodies. Preferably, the ActRII receptor binding molecule antagonizes (e.g., reduces, inhibits, decreases, delays) the binding of a natural ligand to an ActRII receptor. In some embodiments of the disclosed methods, regimens, kits, processes, uses, and compositions, an ActRIIB receptor binding molecule is employed.

[0096] In another embodiment, the composition comprises an anti-ActRII antibody that binds to a binding domain consisting of amino acids 19-134 of SEQ ID NO: 181 (SEQ ID NO: 182), the epitope comprising or consisting of: (a) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188); (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC - SEQ ID NO: 186); (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190); (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR - SEQ ID NO: 189); (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW - SEQ ID NO: 187); (f) amino acids 57-58 of SEQ ID NO: 181 (EQDKR - SEQ ID NO: 189); (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN-SEQ ID NO: 192); or (h) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR).

[0097] In yet another alternative embodiment, the above composition comprises an anti-ActRII antibody that binds to ActRIIB with an affinity that is 10 times or greater than its affinity for ActRIIA.

[0098] In addition, the present disclosure relates to a composition, wherein the anti-ActRIIB antibody comprises: a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14; a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28; a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: 42; a light chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 43 to SEQ ID NO: 56; a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 57 to SEQ ID NO: 70; and a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 71 to SEQ ID NO: 84.

[0099] In certain embodiments, the present disclosure provides a composition wherein the anti-ActRII antibody comprises: (a) a heavy chain variable region CDR1 set forth in SEQ ID NO: 1; a heavy chain variable region CDR2 set forth in SEQ ID NO: 15; a heavy chain variable region CDR3 set forth in SEQ ID NO: 29; a light chain variable region CDR1 set forth in SEQ ID NO: 43; a light chain variable region CDR2 set forth in SEQ ID NO: 57; and a light chain variable region CDR3 set forth in SEQ ID NO: 71, (b) a heavy chain variable region CDR1 set forth in SEQ ID NO: 2; a heavy chain variable region CDR2 set forth in SEQ ID NO: 16; a heavy chain variable region CDR3 set forth in SEQ ID NO: 30; a light chain variable region CDR1 set forth in SEQ ID NO: 44; a light chain variable region CDR2 set forth in SEQ ID NO: 58; and a light chain variable region CDR3 set forth in SEQ ID NO: 72, (c) a heavy chain variable region CDR1 set forth in SEQ ID NO: 3; a heavy chain variable region CDR2 set forth in SEQ ID NO: 17; a light chain variable region CDR2 set forth in SEQ ID NO: 18 NO:31; the heavy chain variable region CDR3 of SEQ ID NO:45; the light chain variable region CDR2 of SEQ ID NO:59; and the light chain variable region CDR3 of SEQ ID NO:73, (d) the heavy chain variable region CDR1 of SEQ ID NO:4; the heavy chain variable region CDR2 of SEQ ID NO:18; the heavy chain variable region CDR3 of SEQ ID NO:32; the light chain variable region CDR1 of SEQ ID NO:46; the light chain variable region CDR2 of SEQ ID NO:60; and the light chain variable region CDR3 of SEQ ID NO:74, (e) the heavy chain variable region CDR1 of SEQ ID NO:5; the heavy chain variable region CDR2 of SEQ ID NO:19; the heavy chain variable region CDR3 of SEQ ID NO:33; the light chain variable region CDR1 of SEQ ID NO:47; the light chain variable region CDR2 of SEQ ID NO:61; and the light chain variable region CDR3 of SEQ ID NO:74. (f) the heavy chain variable region CDR1 set forth in SEQ ID NO:6; the heavy chain variable region CDR2 set forth in SEQ ID NO:20; the heavy chain variable region CDR3 set forth in SEQ ID NO:34; the light chain variable region CDR1 set forth in SEQ ID NO:48; the light chain variable region CDR2 set forth in SEQ ID NO:62; and the light chain variable region CDR3 set forth in SEQ ID NO:76, (g) the heavy chain variable region CDR1 set forth in SEQ ID NO:7;the heavy chain variable region CDR2 shown in SEQ ID NO:21; the heavy chain variable region CDR3 shown in SEQ ID NO:35; the light chain variable region CDR1 shown in SEQ ID NO:49; the light chain variable region CDR2 shown in SEQ ID NO:63; and the light chain variable region CDR3 shown in SEQ ID NO:77, (h) the heavy chain variable region CDR1 shown in SEQ ID NO:8; the heavy chain variable region CDR2 shown in SEQ ID NO:22; the heavy chain variable region CDR3 shown in SEQ ID NO:36; the light chain variable region CDR1 shown in SEQ ID NO:50; the light chain variable region CDR2 shown in SEQ ID NO:64; and the light chain variable region CDR3 shown in SEQ ID NO:78, (i) the heavy chain variable region CDR1 shown in SEQ ID NO:9; the heavy chain variable region CDR2 shown in SEQ ID NO:23; the heavy chain variable region CDR3 shown in SEQ ID NO:37; the light chain variable region CDR1 shown in SEQ ID NO:51; NO:65; and the light chain variable region CDR3 shown in SEQ ID NO:79, (j) the heavy chain variable region CDR1 shown in SEQ ID NO:10; the heavy chain variable region CDR2 shown in SEQ ID NO:24; the heavy chain variable region CDR3 shown in SEQ ID NO:38; the light chain variable region CDR1 shown in SEQ ID NO:52; the light chain variable region CDR2 shown in SEQ ID NO:66; and the light chain variable region CDR3 shown in SEQ ID NO:80, (k) the heavy chain variable region CDR1 shown in SEQ ID NO:11; the heavy chain variable region CDR2 shown in SEQ ID NO:25; the heavy chain variable region CDR3 shown in SEQ ID NO:39; the light chain variable region CDR1 shown in SEQ ID NO:53; the light chain variable region CDR2 shown in SEQ ID NO:67; and the light chain variable region CDR3 shown in SEQ ID NO:81, (i) the heavy chain variable region CDR1 shown in SEQ ID NO:12; the heavy chain variable region CDR2 shown in SEQ ID NO:25; the heavy chain variable region CDR3 shown in SEQ ID NO:39; the light chain variable region CDR1 shown in SEQ ID NO:53; the light chain variable region CDR2 shown in SEQ ID NO:67; and the light chain variable region CDR3 shown in SEQ ID NO:81 NO:26; the heavy chain variable region CDR2 set forth in SEQ ID NO:40; the light chain variable region CDR1 set forth in SEQ ID NO:54; the light chain variable region CDR2 set forth in SEQ ID NO:68; and the light chain variable region CDR3 set forth in SEQ ID NO:82, (m) the heavy chain variable region CDR1 set forth in SEQ ID NO:13; the heavy chain variable region CDR2 set forth in SEQ ID NO:27; the heavy chain variable region CDR3 set forth in SEQ ID NO:41; and the light chain variable region CDR1 set forth in SEQ ID NO:55;The light chain variable region CDR2 set forth in SEQ ID NO:69; and the light chain variable region CDR3 set forth in SEQ ID NO:83, or (n) the heavy chain variable region CDR1 set forth in SEQ ID NO:14; the heavy chain variable region CDR2 set forth in SEQ ID NO:28; the heavy chain variable region CDR3 set forth in SEQ ID NO:42; the light chain variable region CDR1 set forth in SEQ ID NO:56; the light chain variable region CDR2 set forth in SEQ ID NO:70; and the light chain variable region CDR3 set forth in SEQ ID NO:84.

[0100] In another embodiment, the anti-ActRII antibody comprises (i) a full heavy chain amino acid sequence that has at least 95% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160, (ii) a full light chain amino acid sequence that has at least 95% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: 155, or (iii) (a) the variable heavy chain sequence of SEQ ID NO: 99 and the variable light chain sequence of SEQ ID NO: 85; (b) the variable heavy chain sequence of SEQ ID NO: 100 and the variable light chain sequence of SEQ ID NO: 86; (c) the variable heavy chain sequence of SEQ ID NO: 101 and the variable light chain sequence of SEQ ID NO: 87; (d) the variable heavy chain sequence of SEQ ID NO: 102 and the variable light chain sequence of SEQ ID NO: 88; (e) the variable heavy chain sequence of SEQ ID NO: 103 and the variable light chain sequence of SEQ ID NO: 104. NO: 103 and the variable light chain sequence shown in SEQ ID NO: 89; (f) the variable heavy chain sequence shown in SEQ ID NO: 104 and the variable light chain sequence shown in SEQ ID NO: 90; (g) the variable heavy chain sequence shown in SEQ ID NO: 105 and the variable light chain sequence shown in SEQ ID NO: 91; (h) the variable heavy chain sequence shown in SEQ ID NO: 106 and the variable light chain sequence shown in SEQ ID NO: 92; (i) the variable heavy chain sequence shown in SEQ ID NO: 107 and the variable light chain sequence shown in SEQ ID NO: 93; (j) the variable heavy chain sequence shown in SEQ ID NO: 108 and the variable light chain sequence shown in SEQ ID NO: 94; (k) the variable heavy chain sequence shown in SEQ ID NO: 109 and the variable light chain sequence shown in SEQ ID NO: 95; (l) the variable heavy chain sequence shown in SEQ ID NO: 110 and the variable light chain sequence shown in SEQ ID NO: 96; (m) the variable heavy chain sequence shown in SEQ ID NO: 111 and the variable light chain sequence shown in SEQ ID NO: The variable light chain sequence shown in NO:97; or (n) the variable heavy chain sequence shown in SEQ ID NO:112 and the variable light chain sequence shown in SEQ ID NO:98.

[0101] In certain aspects, the present disclosure relates to the above-mentioned compositions, wherein the anti-ActRII antibody comprises (a) a heavy chain sequence set forth in SEQ ID NO: 146 and a light chain sequence set forth in SEQ ID NO: 141; (b) a heavy chain sequence set forth in SEQ ID NO: 147 and a light chain sequence set forth in SEQ ID NO: 142; (c) a heavy chain sequence set forth in SEQ ID NO: 148 and a light chain sequence set forth in SEQ ID NO: 143; (d) a heavy chain sequence set forth in SEQ ID NO: 149 and a light chain sequence set forth in SEQ ID NO: 144; (e) a heavy chain sequence set forth in SEQ ID NO: 150 and a light chain sequence set forth in SEQ ID NO: 145; (f) a heavy chain sequence set forth in SEQ ID NO: 156 and a light chain sequence set forth in SEQ ID NO: 151; (g) a heavy chain sequence set forth in SEQ ID NO: 157 and a light chain sequence set forth in SEQ ID NO: 152; (h) a heavy chain sequence set forth in SEQ ID NO: 158 and a light chain sequence set forth in SEQ ID NO: 153; (i) a heavy chain sequence set forth in SEQ ID NO: 159 and a light chain sequence set forth in SEQ ID NO: 160; The heavy chain sequence shown in SEQ ID NO: 159 and the light chain sequence shown in SEQ ID NO: 154; or (j) the heavy chain sequence shown in SEQ ID NO: 160 and the light chain sequence shown in SEQ ID NO: 155.

[0102] Another subject matter of the present disclosure relates to compositions wherein (i) the anti-ActRII antibody cross-blocks or is cross-blocked by one of the above antibodies, (ii) has altered effector function through mutation of the Fc region, and / or (iii) binds to an epitope recognized by one of the above antibodies.

[0103] In yet another alternative embodiment, the above composition comprises an anti-ActRII antibody that binds to ActRIIB with an affinity that is 10 times or greater than its affinity for ActRIIA.

[0104] In another embodiment, the disclosed composition comprises an anti-ActRII antibody encoded by pBW522 (DSM22873) or pBW524 (DSM22874). Sequence Listing <110> Novartis AG <120> Methods for preventing and treating heart disease <130> 057695 <141> 2018-03-20 <150> US 62 / 476,054 <151> 2017-03-24 <160> 194 <170> PatentIn version 3.5 <210> 1 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 1 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 2 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 3 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 3 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 4 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 4 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 5 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 5 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 6 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 6 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 7 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 7 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 8 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 8 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 9 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 9 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 10 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 10 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 11 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 11 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 12 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 12 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 13 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 13 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 14 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 14 Gly Tyr Thr Phe Thr Ser Ser Tyr Ile Asn 1 5 10 <210> 15 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 15 Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 16 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 16 Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 17 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 17 Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 18 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 18 Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 19 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 19 Met Ile Asn Ala Pro Ile Gly Thr Thr Arg Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 20 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 20 Gln Ile Asn Ala Ala Ser Gly Met Thr Arg Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 21 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 21 Met Ile Asn Ala Pro Ile Gly Thr Thr Arg Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 22 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 22 Thr Ile Asn Pro Val Ser Gly Asn Thr Arg Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 23 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 23 Thr Ile Asn Pro Val Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 24 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 24 Gln Ile Asn Ala Ala Ser Gly Met Thr Arg Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 25 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 25 Asn Ile Asn Ala Ala Ala Gly Ile Thr Leu Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 26 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 26 Thr Ile Asn Pro Pro Thr Gly Gly Thr Tyr Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 27 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 27 Gly Ile Asn Pro Pro Ala Gly Thr Thr Ser Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 28 <211> 17 <212> PRT <213> Artificial <220> <223> CDR <400> 28 Asn Ile Asn Pro Ala Thr Gly His Ala Asp Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 29 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 29 Gly Gly Trp Phe Asp Tyr 1 5 <210> 30 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 30 Gly Gly Trp Phe Asp Tyr 1 5 <210> 31 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 31 Gly Gly Trp Phe Asp Tyr 1 5 <210> 32 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 32 Gly Gly Trp Phe Asp Tyr 1 5 <210> 33 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 33 Gly Gly Trp Phe Asp Tyr 1 5 <210> 34 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 34 Gly Gly Trp Phe Asp Tyr 1 5 <210> 35 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 35 Gly Gly Trp Phe Asp Tyr 1 5 <210> 36 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 36 Gly Gly Trp Phe Asp Tyr 1 5 <210> 37 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 37 Gly Gly Trp Phe Asp Tyr 1 5 <210> 38 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 38 Gly Gly Trp Phe Asp Tyr 1 5 <210> 39 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 39 Gly Gly Trp Phe Asp Tyr 1 5 <210> 40 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 40 Gly Gly Trp Phe Asp Tyr 1 5 <210> 41 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 41 Gly Gly Trp Phe Asp Tyr 1 5 <210> 42 <211> 6 <212> PRT <213> Artificial <220> <223> CDR <400> 42 Gly Gly Trp Phe Asp Tyr 1 5 <210> 43 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 43 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 44 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 44 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 45 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 45 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 46 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 46 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 47 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 47 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 48 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 48 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 49 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 49 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 50 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 50 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 51 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 51 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 52 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 52 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 53 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 53 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 54 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 54 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 55 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 55 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 56 <211> 14 <212> PRT <213> Artificial <220> <223> CDR <400> 56 Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr Asn Tyr Val Asn 1 5 10 <210> 57 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 57 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 58 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 58 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 59 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 59 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 60 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 60 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 61 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 61 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 62 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 62 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 63 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 63 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 64 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 64 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 65 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 65 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 66 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 66 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 67 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 67 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 68 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 68 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 69 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 69 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 70 <211> 11 <212> PRT <213> Artificial <220> <223> CDR <400> 70 Leu Met Ile Tyr Gly Val Ser Lys Arg Pro Ser 1 5 10 <210> 71 <211> 9 <212> PRT <213> Artificial <220> <223> CDR <400> 71 Gln Ala Trp Thr Ser Lys Met Ala Gly 1 5 <210> 72 <211> 9 <212> PRT <213> Artificial <220> <223> CDR <400> 72 Ser Ser Tyr Thr Arg Met Gly His Pro 1 5 <210> 73 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 73 Ala Thr Tyr Gly Lys Gly Val Thr Pro Pro 1 5 10 <210> 74 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 74 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 75 <211> 9 <212> PRT <213> Artificial <220> <223> CDR <400> 75 Gln Ala Trp Thr Ser Lys Met Ala Gly 1 5 <210> 76 <211> 9 <212> PRT <213> Artificial <220> <223> CDR <400> 76 Gln Ala Trp Thr Ser Lys Met Ala Gly 1 5 <210> 77 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 77 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 78 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 78 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 79 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 79 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 80 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 80 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 81 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 81 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 82 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 82 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 83 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 83 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 84 <211> 10 <212> PRT <213> Artificial <220> <223> CDR <400> 84 Gly Thr Phe Ala Gly Gly Ser Tyr Tyr Gly 1 5 10 <210> 85 <211> 112 <212> PRT <213> Artificial <220> <223> VL <400> 85 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Thr Ser Lys 85 90 95 Met Ala Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 <210> 86 <211> 112 <212> PRT <213> Artificial <220> <223> VL <400> 86 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Arg Met 85 90 95 Gly His Pro Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 <210> 87 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 87 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Tyr Gly Lys Gly 85 90 95 Val Thr Pro Pro Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 88 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 88 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 89 <211> 112 <212> PRT <213> Artificial <220> <223> VL <400> 89 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Thr Ser Lys 85 90 95 Met Ala Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 <210> 90 <211> 112 <212> PRT <213> Artificial <220> <223> VL <400> 90 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Thr Ser Lys 85 90 95 Met Ala Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 <210> 91 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 91 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 92 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 92 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 93 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 93 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 94 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 94 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 95 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 95 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 96 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 96 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 97 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 97 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 98 <211> 113 <212> PRT <213> Artificial <220> <223> VL <400> 98 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln <210> 99 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 99 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 100 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 100 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 101 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 101 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 102 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 102 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Asn Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 103 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 103 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asn Ala Pro Ile Gly Thr Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 104 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 104 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gln Ile Asn Ala Ala Ser Gly Met Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 105 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 105 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asn Ala Pro Ile Gly Thr Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 106 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 106 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Asn Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 107 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 107 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 108 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 108 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gln Ile Asn Ala Ala Ser Gly Met Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 109 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 109 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Asn Ala Ala Ala Gly Ile Thr Leu Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 110 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 110 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Pro Thr Gly Gly Thr Tyr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 111 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 111 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Asn Pro Pro Ala Gly Thr Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 112 <211> 115 <212> PRT <213> Artificial <220> <223> VH <400> 112 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Asn Pro Ala Thr Gly His Ala Asp Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 <210> 113 <211> 336 <212> DNA <213> Artificial <220> <223> VL <400> 113 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc caggcttgga cttctaagat ggctggtgtg 300 tttggcggcg gcacgaagtt aaccgttctt ggccag 336 <210> 114 <211> 336 <212> DNA <213> Artificial <220> <223> VL <400> 114 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc tcttcttata ctcgtatggg tcatcctgtg 300 tttggcggcg gcacgaagtt aaccgttctt ggccag 336 <210> 115 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 115 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc gctacttatg gtaagggtgt tactcctcct 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 116 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 116 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 117 <211> 336 <212> DNA <213> Artificial <220> <223> VL <400> 117 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc caggcttgga cttctaagat ggctggtgtg 300 tttggcggcg gcacgaagtt aaccgttctt ggccag 336 <210> 118 <211> 336 <212> DNA <213> Artificial <220> <223> VL <400> 118 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc caggcttgga cttctaagat ggctggtgtg 300 tttggcggcg gcacgaagtt aaccgttctt ggccag 336 <210> 119 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 119 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 120 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 120 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 121 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 121 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 122 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 122 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 123 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 123 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 124 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 124 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtactg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 125 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 125 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 126 <211> 339 <212> DNA <213> Artificial <220> <223> VL <400> 126 gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339 <210> 127 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 127 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact atcaatccgg tttctggcaa tacgtcttac 180 gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 128 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 128 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact atcaatccgg tttctggcaa tacgtcttac 180 gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 129 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 129 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact atcaatccgg tttctggcaa tacgtcttac 180 gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 130 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 130 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact atcaatccgg tttctggcaa tacgtcttac 180 gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 131 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 131 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcatg attaatgctc ctattggtac tactcgttat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 132 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 132 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggccag attaatgctg cttctggtat gactcgttat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 133 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 133 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcatg attaatgctc ctattggtac tactcgttat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 134 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 134 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact atcaatccgg tttctggcaa tacgcgttac 180 gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 135 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 135 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact atcaatccgg tttctggctc tacgtcttac 180 gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 136 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 136 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggccag attaatgctg cttctggtat gactcgttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 137 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 137 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcaat attaatgctg ctgctggtat tactctttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 138 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 138 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcact attaatcctc ctactggagg tacttattat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 139 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 139 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcggt attaatcctc ctgctggtac tacttcttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 140 <211> 345 <212> DNA <213> Artificial <220> <223> VH <400> 140 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcaat attaatcctg ctactggtca tgctgattat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctca 345 <210> 141 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 141 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 142 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 142 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 143 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 143 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 144 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 144 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 145 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 145 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 146 <211> 445 <212> PRT <213> Artificial <220> <223> heavy chain <400> 146 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 147 <211> 445 <212> PRT <213> Artificial <220> <223> heavy chain <400> 147 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gln Ile Asn Ala Ala Ser Gly Met Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 148 <211> 445 <212> PRT <213> Artificial <220> <223> heavy chain <400> 148 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Asn Ala Ala Ala Gly Ile Thr Leu Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 149 <211> 445 <212> PRT <213> Artificial <220> <223> heavy chain <400> 149 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Asn Pro Pro Ala Gly Thr Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 150 <211> 445 <212> PRT <213> Artificial <220> <223> heavy chain <400> 150 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Asn Pro Ala Thr Gly His Ala Asp Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 151 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 151 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 152 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 152 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 153 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 153 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 154 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 154 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 155 <211> 217 <212> PRT <213> Artificial <220> <223> light chain <400> 155 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 156 <211> 441 <212> PRT <213> Artificial <220> <223> heavy chain <400> 156 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly 180 185 190 Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys 210 215 220 Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 225 230 235 240 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 245 250 255 Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 260 265 270 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 275 280 285 Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His 290 295 300 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 305 310 315 320 Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln 325 330 335 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 340 345 350 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 355 360 365 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 370 375 380 Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu 385 390 395 400 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 405 410 415 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 420 425 430 Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 157 <211> 441 <212> PRT <213> Artificial <220> <223> heavy chain <400> 157 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gln Ile Asn Ala Ala Ser Gly Met Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly 180 185 190 Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys 210 215 220 Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 225 230 235 240 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 245 250 255 Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 260 265 270 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 275 280 285 Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His 290 295 300 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 305 310 315 320 Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln 325 330 335 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 340 345 350 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 355 360 365 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 370 375 380 Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu 385 390 395 400 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 405 410 415 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 420 425 430 Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 158 <211> 441 <212> PRT <213> Artificial <220> <223> heavy chain <400> 158 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Asn Ala Ala Ala Gly Ile Thr Leu Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly 180 185 190 Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys 210 215 220 Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 225 230 235 240 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 245 250 255 Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 260 265 270 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 275 280 285 Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His 290 295 300 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 305 310 315 320 Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln 325 330 335 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 340 345 350 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 355 360 365 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 370 375 380 Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu 385 390 395 400 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 405 410 415 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 420 425 430 Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 159 <211> 441 <212> PRT <213> Artificial <220> <223> heavy chain <400> 159 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Asn Pro Pro Ala Gly Thr Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly 180 185 190 Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys 210 215 220 Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 225 230 235 240 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 245 250 255 Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 260 265 270 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 275 280 285 Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His 290 295 300 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 305 310 315 320 Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln 325 330 335 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 340 345 350 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 355 360 365 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 370 375 380 Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu 385 390 395 400 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 405 410 415 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 420 425 430 Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 160 <211> 441 <212> PRT <213> Artificial <220> <223> heavy chain <400> 160 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Asn Pro Ala Thr Gly His Ala Asp Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115 120 125 Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly 180 185 190 Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys 195 200 205 Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys 210 215 220 Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 225 230 235 240 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 245 250 255 Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 260 265 270 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 275 280 285 Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His 290 295 300 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 305 310 315 320 Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln 325 330 335 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 340 345 350 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 355 360 365 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 370 375 380 Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu 385 390 395 400 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 405 410 415 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 420 425 430 Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 161 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 161 cagagcgccc tgacccagcc cgccagcgtg tccggcagcc caggccagtc tatcacaatc 60 agctgcaccg gcacctccag cgacgtgggc agctacaact acgtgaactg gtatcagcag 120 caccccggca aggcccccaa gctgatgatc tacggcgtga gcaagaggcc cagcggcgtg 180 tccaacaggt tcagcggcag caagagcggc aacaccgcca gcctgacaat cagtgggctg 240 caggctgagg acgaggccga ctactactgc ggcacctttg ccggcggatc atactacggc 300 gtgttcggcg gagggaccaa gctgaccgtg ctgggccagc ctaaggctgc ccccagcgtg 360 accctgttcc cccccagcag cgaggagctg caggccaaca aggccaccct ggtgtgcctg 420 atcagcgact tctacccagg cgccgtgacc gtggcctgga aggccgacag cagccccgtg 480 aaggccggcg tggagaccac cacccccagc aagcagagca acaacaagta cgccgccagc 540 agctacctga gcctgacccc cgagcagtgg aagagccaca ggtcctacag ctgccaggtg 600 acccacgagg gcagcaccgt ggaaaagacc gtggccccaa ccgagtgcag c 651 <210> 162 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 162 cagagcgcc tgacccagcc cgccagcgtg tccggcagcc caggccagtc tatcacaatc 60 agctgcaccg gcacctccag cgacgtgggc agctacaact acgtgaactg gtatcagcag 120 caccccggca aggcccccaa gctgatgatc tacggcgtga gcaagaggcc cagcggcgtg 180 tccaacaggt tcagcggcag caagagcgg aacaccgcca gcctgacaat cagtgggctg 240 caggctgagg acgaggccga ctactactgc ggcacctttg ccggcggatc atactacggc 300 gtgttcggcg gaggaccaa gctgaccgtg ctgggccagc ctaaggctgc ccccagcgtg 360 accctgttcc cccccagcag cgaggagctg caggccaaca aggccaccct ggtgtgcctg 420 atcagcgact tctacccagg cgccgtgacc gtggcctgga aggccgacag cagccccgtg 480 aaggccggcg tggagaccac cacccccagc aagcagagca acaacaagta cgccgccagc 540 agctacctga gcctgacccc cgagcagtgg aagagccaca ggtcctacag ctgccaggtg 600 acccacgagg gcagcaccgt ggaaaagacc gtggccccaa ccgagtgcag c 651 <210> 163 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 163 cagagcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcagc ccaaggctgc cccctcggtc 360 actctgttcc cgccctcctc tgaggagctt caagccaaca aggccacact ggtgtgtctc 420 ataagtgact tctacccggg agccgtgaca gtggcctgga aggcagatag cagccccgtc 480 aaggcgggag tggagaccac cacaccctcc aaacaaagca acaacaagta cgcggccagc 540 agctatctga gcctgacgcc tgagcagtgg aagtcccaca gaagctacag ctgccaggtc 600 acgcatgaag ggagcaccgt ggagaagaca gtggccccta cagaatgttc a 651 <210> 164 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 164 cagagcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcagc ccaaggctgc cccctcggtc 360 actctgttcc cgccctcctc tgaggagctt caagccaaca aggccacact ggtgtgtctc 420 ataagtgact tctacccggg agccgtgaca gtggcctgga aggcagatag cagccccgtc 480 aaggcgggag tggagaccac cacaccctcc aaacaaagca acaacaagta cgcggccagc 540 agctatctga gcctgacgcc tgagcagtgg aagtcccaca gaagctacag ctgccaggtc 600 acgcatgaag ggagcaccgt ggagaagaca gtggccccta cagaatgttc a 651 <210> 165 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 165 cagagcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcagc ccaaggctgc cccctcggtc 360 actctgttcc cgccctcctc tgaggagctt caagccaaca aggccacact ggtgtgtctc 420 ataagtgact tctacccggg agccgtgaca gtggcctgga aggcagatag cagccccgtc 480 aaggcgggag tggagaccac cacaccctcc aaacaaagca acaacaagta cgcggccagc 540 agctatctga gcctgacgcc tgagcagtgg aagtcccaca gaagctacag ctgccaggtc 600 acgcatgaag ggagcaccgt ggagaagaca gtggccccta cagaatgttc a 651 <210> 166 <211> 1335 <212> DNA <213> Artificial <220> <223> heavy chain <400> 166 caggtgcagc tggtgcagag cggagctgag gtgaagaagc caggcgccag cgtcaaggtg 60 tcctgcaagg ccagcggcta caccttcacc agcagctaca tcaactgggt ccgccaggct 120 cctgggcagg gactggagtg gatgggcacc atcaaccccg tgtccggcag caccagctac 180 gcccagaagt tccagggcag agtcaccatg accagggaca ccagcatcag caccgcctac 240 atggagctgt ccaggctgag aagcgacgac accgccgtgt actactgcgc caggggcggc 300 tggttcgact actggggcca gggcaccctg gtgaccgtgt cctcagctag caccaagggc 360 cccagcgtgt tccccctggc ccccagcagc aagagcacct ccggcggcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag cccgtgaccg tgtcctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctccaga gcagcggcct gtacagcctg 540 tccagcgtgg tgacagtgcc cagcagcagc ctgggcacc agacctacat ctgcaacgtg 600 aaccacaagc ccagcaacac caaggtggac aagagagtgg agcccaagag ctgcgacaag 660 acccacacct gccccccctg cccagcccc gaagctgcag gcggcccttc cgtgttcctg 720 ttcccccccca agcccaagga caccctgatg atcagcagga cccccgaggt gacctgcgtg 780 gtggtggacg tgagccacga ggacccagag gtgaagttca actggtacgt ggacggcgtg 840 gaggtgcaca acgccaagac caagcccaga gaggcagt acaacagcac ctacagggtg gtgtccgtgc tgaccgtgct gcaccaggac tggctgaacg gcaaagaata caagtgcaag gtctccaaca aggccctgcc tgcccccatc gaaaagacca tcagcaaggc caagggccag cccgggagc cccaggtgta caccctgccc ccttctcggg aggagatgac caagaccag gtgtccctga cctgtctggt gaagggcttc taccccagcg acatcgccgt ggagtggggag agcaacggcc agcccgagaa caactacaag accacccccc cagtgctgga cagcgacggc agcttcttcc tgtacagcaa gctgaccgtg gacaagagca ggtggcagca gggcaacgtg ttcagctgca gcgtgatgca cgaggccctg cacaaccact acacccagaa gagcctgagc ctgtcacccg gcaag <210> 167 <211> 1335 <212> DNA <213> Artificial <220> <223> heavy chain <400> 167 60. caggtgcagc tggtgcagag cggagctgag gtgaagaagc caggcgccag cgtcaaggtg tcctgcaagg ccagcggcta caccttcacc agcagctaca tcaactgggt gcgccaggct 120 ccagggcagg gactggagtg gatgggccag atcaacgccg ccagcggcat gaccagatac 180 gcccagaagt tccagggcag agtcacaatg accagggaca cctctatcag caccgcctac 240 atggagctgt ccaggctgag aagcgacgac accgccgtgt actactgcgc caggggcggc 300 tggttcgact actggggcca gggcaccctg gtgaccgtgt cctcagctag caccaagggc 360 cccagcgtgt tccccctggc ccccagcagc aagagcacct ccggcggcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag cccgtgaccg tgtcctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctccaga gcagcggcct gtacagcctg 540 tccagcgtgg tgacagtgcc cagcagcagc ctgggcacc agacctacat ctgcaacgtg 600 aaccacaagc ccagcaacac caaggtggac aagagagtgg agcccaagag ctgcgacaag 660 acccacacct gccccccctg cccagcccc gaagctgcag gcggcccttc cgtgttcctg 720 ttcccccccca agcccaagga caccctgatg atcagcagga cccccgaggt gacctgcgtg 780 gtggtggacg tgagccacga ggacccagag gtgaagttca actggtacgt ggacggcgtg 840 gaggtgcaca acgccaagac caagcccaga gaggcagt acaacagcac ctacagggtg gtgtccgtgc tgaccgtgct gcaccaggac tggctgaacg gcaaagaata caagtgcaag gtctccaaca aggccctgcc tgcccccatc gaaaagacca tcagcaaggc caagggccag cccgggagc cccaggtgta caccctgccc ccttctcggg aggagatgac caagaccag gtgtccctga cctgtctggt gaagggcttc taccccagcg acatcgccgt ggagtggggag agcaacggcc agcccgagaa caactacaag accacccccc cagtgctgga cagcgacggc agcttcttcc tgtacagcaa gctgaccgtg gacaagagca ggtggcagca gggcaacgtg ttcagctgca gcgtgatgca cgaggccctg cacaaccact acacccagaa gagcctgagc ctgtcacccg gcaag <210> 168 <211> 1335 <212> DNA <213> Artificial <220> <223> heavy chain <400> 168 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcaat attaatgctg ctgctggtat tactctttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gccgcctgcg tagcgatgat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctcagcctc caccaagggt 360 ccatcggtct tccccctggc accctcctcc aagagcacct ctgggggcac agcggccctg 420 ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgcc 480 ctgaccagcg gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc 540 agcagcgtgg tgaccgtgcc ctccagcagc ttgggcaccc agacctacat ctgcaacgtg 600 aatcacaagc ccagcaacac caaggtggac aagagagttg agcccaaatc ttgtgacaaa 660 actcacacat gcccaccgtg cccagcacct gaagcagcgg ggggaccgtc agctttcctc 720 ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 840 900. gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgggtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 960 gtctccaaca aagccctccc agcccccatc gagaaacca tctccaaagc caaagggcag ccccgagac cacaggtgta caccctgccc ccatcccggg aggagatgac caagaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtggggag agcaatgggc agccggagaa caactacag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaa <210> 169 <211> 1335 <212> DNA <213> Artificial <220> <223> heavy chain <400> 169 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcggt attaatcctc ctgctggtac tacttcttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gccgcctgcg tagcgatgat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctcagcctc caccaagggt 360 ccatcggtct tccccctggc accctcctcc aagagcacct ctgggggcac agcggccctg 420 ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgcc 480 ctgaccagcg gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc 540 agcagcgtgg tgaccgtgcc ctccagcagc ttgggcaccc agacctacat ctgcaacgtg 600 aatcacaagc ccagcaacac caaggtggac aagagagttg agcccaaatc ttgtgacaaa 660 actcacacat gcccaccgtg cccagcacct gaagcagcgg ggggaccgtc agtcttcctc 720 ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 840 900. gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgggtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 960 gtctccaaca aagccctccc agcccccatc gagaaacca tctccaaagc caaagggcag ccccgagac cacaggtgta caccctgccc ccatcccggg aggagatgac caagaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtggggag agcaatgggc agccggagaa caactacag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaa <210> 170 <211> 1335 <212> DNA <213> Artificial <220> <223> heavy chain <400> 170 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcaat attaatcctg ctactggtca tgctgattat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gccgcctgcg tagcgatgat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctcagcctc caccaagggt 360 ccatcggtct tccccctggc accctcctcc aagagcacct ctgggggcac agcggccctg 420 ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgcc 480 ctgaccagcg gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc 540 agcagcgtgg tgaccgtgcc ctccagcagc ttgggcaccc agacctacat ctgcaacgtg 600 aatcacaagc ccagcaacac caaggtggac aagagagttg agcccaaatc ttgtgacaaa 660 actcacacat gcccaccgtg cccagcacct gaagcagcgg ggggaccgtc agtcttcctc 720 ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 840 900. gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgggtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 960 gtctccaaca aagccctccc agcccccatc gagaaacca tctccaaagc caaagggcag ccccgagac cacaggtgta caccctgccc ccatcccggg aggagatgac caagaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtggggag agcaatgggc agccggagaa caactacag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaa <210> 171 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 171 cagagcgcc tgacccagcc cgccagcgtg tccggcagcc caggccagtc tatcacaatc 60 agctgcaccg gcacctccag cgacgtgggc agctacaact acgtgaactg gtatcagcag 120 caccccggca aggcccccaa gctgatgatc tacggcgtga gcaagaggcc cagcggcgtg 180 tccaacaggt tcagcggcag caagagcgg aacaccgcca gcctgacaat cagtgggctg 240 caggctgagg acgaggccga ctactactgc ggcacctttg ccggcggatc atactacggc 300 gtgttcggcg gaggaccaa gctgaccgtg ctgggccagc ctaaggctgc ccccagcgtg 360 accctgttcc ccccagcag cgaggagctg caggccaaca aggccaccct ggtgtgcctg 420 atcagcgact tctacccagg cgccgtgacc gtggcctgga aggccgacag cagccccgtg 480 aaggccggcg tggagaccac cacccccagc aagcagagca acaacaagta cgccgccagc 540 agctacctga gcctgacccc cgagcagtgg aagagccaca ggtcctacag ctgccaggtg 600 acccacgagg gcagcaccgt ggaaaagacc gtggccccaa ccgagtgcag c 651 <210> 172 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 172 cagagcgcc tgacccagcc cgccagcgtg tccggcagcc caggccagtc tatcacaatc 60 agctgcaccg gcacctccag cgacgtgggc agctacaact acgtgaactg gtatcagcag 120 caccccggca aggcccccaa gctgatgatc tacggcgtga gcaagaggcc cagcggcgtg 180 tccaacaggt tcagcggcag caagagcgg aacaccgcca gcctgacaat cagtgggctg 240 caggctgagg acgaggccga ctactactgc ggcacctttg ccggcggatc atactacggc 300 gtgttcggcg gaggaccaa gctgaccgtg ctgggccagc ctaaggctgc ccccagcgtg 360 accctgttcc ccccagcag cgaggagctg caggccaaca aggccaccct ggtgtgcctg 420 atcagcgact tctacccagg cgccgtgacc gtggcctgga aggccgacag cagccccgtg 480 aaggccggcg tggagaccac cacccccagc aagcagagca acaacaagta cgccgccagc 540 agctacctga gcctgacccc cgagcagtgg aagagccaca ggtcctacag ctgccaggtg 600 acccacgagg gcagcaccgt ggaaaagacc gtggccccaa ccgagtgcag c 651 <210> 173 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 173 cagagcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcagc ccaaggctgc cccctcggtc 360 actctgttcc cgccctcctc tgaggagctt caagccaaca aggccacact ggtgtgtctc 420 ataagtgact tctacccggg agccgtgaca gtggcctgga aggcagatag cagccccgtc 480 aaggcgggag tggagaccac cacaccctcc aaacaaagca acaacaagta cgcggccagc 540 agctatctga gcctgacgcc tgagcagtgg aagtcccaca gaagctacag ctgccaggtc 600 acgcatgaag ggagcaccgt ggagaagaca gtggccccta cagaatgttc a 651 <210> 174 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 174 cagagcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcagc ccaaggctgc cccctcggtc 360 actctgttcc cgccctcctc tgaggagctt caagccaaca aggccacact ggtgtgtctc 420 ataagtgact tctacccggg agccgtgaca gtggcctgga aggcagatag cagccccgtc 480 aaggcgggag tggagaccac cacaccctcc aaacaaagca acaacaagta cgcggccagc 540 agctatctga gcctgacgcc tgagcagtgg aagtcccaca gaagctacag ctgccaggtc 600 acgcatgaag ggagcaccgt ggagaagaca gtggccccta cagaatgttc a 651 <210> 175 <211> 651 <212> DNA <213> Artificial <220> <223> light chain <400> 175 cagagcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60 tcgtgtacgg gtactagcag cgatgttggt tcttataatt atgtgaattg gtaccagcag 120 catcccggga aggcgccgaa acttatgatt tatggtgttt ctaagcgtcc ctcaggcgtg 180 agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240 caagcggaag acgaagcgga ttattattgc ggtacttttg ctggtggttc ttattatggt 300 gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcagc ccaaggctgc cccctcggtc 360 actctgttcc cgccctcctc tgaggagctt caagccaaca aggccacact ggtgtgtctc 420 ataagtgact tctacccggg agccgtgaca gtggcctgga aggcagatag cagccccgtc 480 aaggcgggag tggagaccac cacaccctcc aaacaaagca acaacaagta cgcggccagc 540 agctatctga gcctgacgcc tgagcagtgg aagtcccaca gaagctacag ctgccaggtc 600 acgcatgaag ggagcaccgt ggagaagaca gtggccccta cagaatgttc a 651 <210> 176 <211> 1323 <212> DNA <213> Artificial <220> <223> heavy chain <400> 176 caggtgcagc tggtgcagag cggagctgag gtgaagaagc caggcgccag cgtcaaggtg 60 tcctgcaagg ccagcggcta caccttcacc agcagctaca tcaactgggt ccgccaggct 120 cctgggcagg gactggagtg gatgggcacc atcaaccccg tgtccggcag caccagctac 180 gcccagaagt tccagggcag agtcaccatg accagggaca ccagcatcag caccgcctac 240 atggagctgt ccaggctgag aagcgacgac accgccgtgt actactgcgc caggggcggc 300 tggttcgact actggggcca gggcaccctg gtgaccgtgt cctcagctag caccaagggc 360 cccagcgtgt tccccctggc cccctgcagc agaagcacca gcgagagcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag ccagtgaccg tgtcctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct gtacagcctg 540 tccagcgtgg tgaccgtgcc cagcagcaac ttcggcaccc agacctacac ctgcaacgtg 600 gaccacaagc ccagcaacac caaggtggac aagaccgtgg agaggaagtg ctgcgtggag 660 tgccccccct gcccagcccc cccagtggcc ggaccctccg tgttcctgtt cccccccaag 720 cccaaggaca ccctgatgat cagcaggacc cccgaggtga cctgcgtggt ggtggacgtg 780 agccacgagg acccagaggt gcagttcaac tggtacgtgg acggcgtgga ggtgcacaac 840 gccaagacca agcccagaga ggaacagttt aacagcacct tcagggtggt gtccgtgctg 900 accgtggtgc accaggactg gctgaacggc aaagagtaca agtgcaaggt ctccaacaag 960 ggcctgccag cccccatcga gaaaaccatc agcaagacca agggccagcc acgggagccc 1020 caggtgtaca ccctgccccc cagccgggag gaaatgacca agaaccaggt gtccctgacc 1080 tgtctggtga agggcttcta ccccagcgac atcgccgtgg agtgggagag caacggccag 1140 cccgagaaca actacaagac cacccccccc atgctggaca gcgacggcag cttcttcctg 1200 tacagcaagc tgacagtgga caagagcagg tggcagcagg gcaacgtgtt cagctgcagc 1260 gtgatgcacg aggccctgca caaccactac acccaagaga gcctgagcct gtcccccggc 1320 area 1323 <210> 177 <211> 1323 <212> DNA <213> Artificial <220> <223> heavy chain <400> 177 caggtgcagc tggtgcagag cggagctgag gtgaagaagc caggcgccag cgtcaaggtg 60 tcctgcaagg ccagcggcta caccttcacc agcagctaca tcaactgggt gcgccaggct 120 ccagggcagg gactggagtg gatgggccag atcaacgccg ccagcggcat gaccagatac 180 gcccagaagt tccagggcag agtcacaatg accagggaca cctctatcag caccgcctac 240 atggagctgt ccaggctgag aagcgacgac accgccgtgt actactgcgc caggggcggc 300 tggttcgact actggggcca gggcaccctg gtgaccgtgt cctcagctag caccaagggc 360 cccagcgtgt tccccctggc cccctgcagc agaagcacca gcgagagcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag ccagtgaccg tgtcctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct gtacagcctg 540 tccagcgtgg tgaccgtgcc cagcagcaac ttcggcaccc agacctacac ctgcaacgtg 600 gaccacaagc ccagcaacac caaggtggac aagaccgtgg agaggaagtg ctgcgtggag 660 tgccccccct gcccagcccc cccagtggcc ggaccctccg tgttcctgtt cccccccaag 720 cccaaggaca ccctgatgat cagcaggacc cccgaggtga cctgcgtggt ggtggacgtg 780 agccacgagg acccagaggt gcagttcaac tggtacgtgg acggcgtgga ggtgcacaac 840 gccaagacca agcccagaga ggaacagttt aacagcacct tcagggtggt gtccgtgctg 900 accgtggtgc accaggactg gctgaacggc aaagagtaca agtgcaaggt ctccaacaag 960 ggcctgccag cccccatcga gaaaaccatc agcaagacca agggccagcc acgggagccc 1020 caggtgtaca ccctgccccc cagccgggag gaaatgacca agaaccaggt gtccctgacc 1080 tgtctggtga agggcttcta ccccagcgac atcgccgtgg agtgggagag caacggccag 1140 cccgagaaca actacaagac cacccccccc atgctggaca gcgacggcag cttcttcctg 1200 tacagcaagc tgacagtgga caagagcagg tggcagcagg gcaacgtgtt cagctgcagc 1260 gtgatgcacg aggccctgca caaccactac acccagaaga gcctgagcct gtcccccggc 1320 Aug 1323 <210> 178 <211> 1323 <212> DNA <213> Artificial <220> <223> heavy chain <400> 178 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcaat attaatgctg ctgctggtat tactctttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gccgcctgcg tagcgatgat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctcagcttc caccaagggc 360 cccagcgtgt tccccctggc cccctgcagc agaagcacca gcgagagcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag cccgtgaccg tgagctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct gtacagcctg 540 agcagcgtgg tgaccgtgcc cagcagcaac ttcggcaccc agacctacac ctgcaacgtg 600 gaccacaagc ccagcaacac caaggtggac aagaccgtgg agcggaagtg ctgcgtggag 660 tgccccccct gccctgcccc tcctgtggcc ggaccctccg tgttcctgtt cccccccaag 720 cccaaggaca ccctgatgat cagccggacc cccgaggtga cctgcgtggt ggtggacgtg 780 agccacgagg accccgaggt gcagttcaac tggtacgtgg acggcgtgga ggtgcacaac 840 gccaagacca agccccggga ggaacagttc aacagcacct tccgggtggt gtccgtgctg 900 accgtggtgc accaggactg gctgaacggc aaagaataca agtgcaaggt gtccaacaag 960 ggcctgcctg cccccatcga gaaaaccatc agcaagacaa agggccagcc cagggaaccc 1020 caggtgtaca ccctgccccc cagccgggag gaaatgacca agaaccaggt gtccctgacc 1080 tgtctggtga agggcttcta ccccagcgac atcgccgtgg agtgggagag caacggccag 1140 cccgagaaca actacaagac cacccccccc atgctggaca gcgacggcag cttcttcctg 1200 tacagcaagc tgacagtgga caagagccgg tggcagcagg gcaacgtgtt cagctgcagc 1260 gtgatgcacg aggccctgca caaccactac acccagaaga gcctgagcct gtcccccggc 1320 aaa 1323 <210> 179 <211> 1323 <212> DNA <213> Artificial <220> <223> heavy chain <400> 179 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcggt attaatcctc ctgctggtac tacttcttat 180 gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gccgcctgcg tagcgatgat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctcagcttc caccaagggc 360 cccagcgtgt tccccctggc cccctgcagc agaagcacca gcgagagcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag cccgtgaccg tgagctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct gtacagcctg 540 agcagcgtgg tgaccgtgcc cagcagcaac ttcggcaccc agacctacac ctgcaacgtg 600 gaccacaagc ccagcaacac caaggtggac aagaccgtgg agcggaagtg ctgcgtggag 660 tgccccccct gccctgcccc tcctgtggcc ggaccctccg tgttcctgtt cccccccaag 720 cccaaggaca ccctgatgat cagccggacc cccgaggtga cctgcgtggt ggtggacgtg 780 agccacgagg accccgaggt gcagttcaac tggtacgtgg acggcgtgga ggtgcacaac 840 gccaagacca agccccggga ggaacagttc aacagcacct tccgggtggt gtccgtgctg 900 accgtggtgc accaggactg gctgaacggc aaagaataca agtgcaaggt gtccaacaag 960 ggcctgcctg cccccatcga gaaaaccatc agcaagacaa agggccagcc cagggaaccc 1020 caggtgtaca ccctgccccc cagccgggag gaaatgacca agaaccaggt gtccctgacc 1080 tgtctggtga agggcttcta ccccagcgac atcgccgtgg agtgggagag caacggccag 1140 cccgagaaca actacaagac cacccccccc atgctggaca gcgacggcag cttcttcctg 1200 tacagcaagc tgacagtgga caagagccgg tggcagcagg gcaacgtgtt cagctgcagc 1260 gtgatgcacg aggccctgca caaccactac acccagaaga gcctgagcct gtcccccggc 1320 aaa 1323 <210> 180 <211> 1323 <212> DNA <213> Artificial <220> <223> heavy chain <400> 180 caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60 agctgcaaag cctccggata tacctttact tcttcttata ttaattgggt ccgccaagcc 120 cctgggcagg gtctcgagtg gatgggcaat attaatcctg ctactggtca tgctgattat 180 gctcagaagt ttcagggtcg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240 atggaactga gccgcctgcg tagcgatgat acggccgtgt attattgcgc gcgtggtggt 300 tggttgatt attggggcca aggcaccctg gtgacggtta gctcagcttc caccaagggc 360 cccagcgtgt tccccctggc cccctgcagc agaagcacca gcgagagcac agccgccctg 420 ggctgcctgg tgaaggacta cttccccgag cccgtgaccg tgagctggaa cagcggagcc 480 ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct gtacagcctg 540 agcagcgtgg tgaccgtgcc cagcagcaac ttcggcaccc agacctacac ctgcaacgtg 600 gaccacaagc ccagcaacac caaggtggac aagaccgtgg agcggaagtg ctgcgtggag 660 tgccccccct gccctgcccc tcctgtggcc ggaccctccg tgttcctgtt cccccccaag 720 cccaaggaca ccctgatgat cagccggacc cccgaggtga cctgcgtggt ggtggacgtg 780 agccacgagg accccgaggt gcagttcaac tggtacgtgg acggcgtgga ggtgcacaac 840 gccaagacca agccccggga ggaacagttc aacagcacct tccgggtggt gtccgtgctg 900 accgtggtgc accaggactg gctgaacggc aaagaataca agtgcaaggt gtccaacaag 960 ggcctgcctg cccccatcga gaaaaccatc agcaagacaa agggccagcc cagggaaccc 1020 caggtgtaca ccctgccccc cagccgggag gaaatgacca agaaccaggt gtccctgacc 1080 tgtctggtga agggctctta ccccagcgac atcgccgtgg agtgggag caacggccag 1140 cccgagacaca actacagac cacccccccc atgctggaca gcgacggcag cttctctcctg 1200 tacagcaagc tgacagtgga tacagccgg tggcagcagg gcaacgtgtt cagctgcagc 1260 gtgatgcacg aggccctgca caccactac acccagaga gcctgagcct gtcccccggc 1320 aaa 1323 <210> 181 <211> 512 <212> PRT <213> Homo sapiens <400> 181 Met Thr Ala Pro Trp Val Leu Ala Leu Ala Leu Trp Gly Ser Leu Cys 1 5 10 15 Ala Gly Ser Gly Arg Gly Glu Ala Glu Thr Arg Glu Cys Ile Tyr Tyr 20 25 30 Asn Only Asn Terp Glu Leu Glu Arg Thr Gln Ser Gly Asn Leu Glu Arg 35 40 45 Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys Tyr Ala Is Trp Arg 50 55 60 Asn Ser Gly Thr Ile Glu Leu Val Lys Gly Cys Trp Leu Asp 65 70 75 80 Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val Ala Thr Glu Glu Asn 85 90 95 Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn Phe Cys Asn Glu Arg 100 105 110 Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu Val Thr Tyr Glu Pro 115 120 125 Pro Pro Thr Ala Pro Thr Leu Leu Thr Val Leu Ala Tyr Ser Leu Leu 130 135 140 Pro Ile Gly Gly Leu Ser Leu Ile Val Leu Leu Ala Phe Trp Met Tyr 145 150 155 160 Arg His Arg Lys Pro Pro Tyr Gly His Val Asp Ile His Glu Asp Pro 165 170 175 Gly Pro Pro Pro Pro Ser Pro Leu Val Gly Leu Lys Pro Leu Gln Leu 180 185 190 Leu Glu Ile Lys Ala Arg Gly Arg Phe Gly Cys Val Trp Lys Ala Gln 195 200 205 Leu Met Asn Asp Phe Val Ala Val Lys Ile Phe Pro Leu Gln Asp Lys 210 215 220 Gln Ser Trp Gln Ser Glu Arg Glu Ile Phe Ser Thr Pro Gly Met Lys 225 230 235 240 His Glu Asn Leu Leu Gln Phe Ile Ala Ala Glu Lys Arg Gly Ser Asn 245 250 255 Leu Glu Val Glu Leu Trp Leu Ile Thr Ala Phe His Asp Lys Gly Ser 260 265 270 Leu Thr Asp Tyr Leu Lys Gly Asn Ile Ile Thr Trp Asn Glu Leu Cys 275 280 285 His Val Ala Glu Thr Met Ser Arg Gly Leu Ser Tyr Leu His Glu Asp 290 295 300 Val Pro Trp Cys Arg Gly Glu Gly His Lys Pro Ser Ile Ala His Arg 305 310 315 320 Asp Phe Lys Ser Lys Asn Val Leu Leu Lys Ser Asp Leu Thr Ala Val 325 330 335 Leu Ala Asp Phe Gly Leu Ala Val Arg Phe Glu Pro Gly Lys Pro Pro 340 345 350 Gly Asp Thr His Gly Gln Val Gly Thr Arg Arg Tyr Met Ala Pro Glu 355 360 365 Val Leu Glu Gly Ala Ile Asn Phe Gln Arg Asp Ala Phe Leu Arg Ile 370 375 380 Asp Met Tyr Ala Met Gly Leu Val Leu Trp Glu Leu Val Ser Arg Cys 385 390 395 400 Lys Ala Ala Asp Gly Pro Val Asp Glu Tyr Met Leu Pro Phe Glu Glu 405 410 415 Glu Ile Gly Gln His Pro Ser Leu Glu Glu Leu Gln Glu Val Val Val 420 425 430 His Lys Lys Met Arg Pro Thr Ile Lys Asp His Trp Leu Lys His Pro 435 440 445 Gly Leu Ala Gln Leu Cys Val Thr Ile Glu Ala Cys Trp Asp His Asp 450 455 460 Ala Glu Ala Arg Leu Ser Ala Gly Cys Val Glu Glu Arg Val Ser Leu 465 470 475 480 Ile Arg Arg Ser Val Asn Gly Thr Thr Ser Asp Cys Leu Val Ser Leu 485 490 495 Val Thr Ser Val Thr Asn Val Asp Leu Pro Pro Lys Glu Ser Ser Ile 500 505 510 <210> 182 <211> 116 <212> PRT <213> Homo sapiens <400> 182 Ser Gly Arg Gly Glu Ala Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala 1 5 10 15 Asn Trp Glu Leu Glu Arg Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu 20 25 30 Gly Glu Gln Asp Lys Arg Leu His Cys Tyr Ala Ser Trp Arg Asn Ser 35 40 45 Ser Gly Thr Ile Glu Leu Val Lys Lys Gly Cys Trp Leu Asp Asp Phe 50 55 60 Asn Cys Tyr Asp Arg Gln Glu Cys Val Ala Thr Glu Glu Asn Pro Gln 65 70 75 80 Val Tyr Phe Cys Cys Cys Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr 85 90 95 His Leu Pro Glu Ala Gly Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro 100 105 110 Thr Ala Pro Thr 115 <210> 183 <211> 15 <212> PRT <213> Homo sapiens <400> 183 Ile Glu Leu Val Lys Lys Gly Ser Trp Leu Asp Asp Phe Asn Ser 1 5 10 15 <210> 184 <211> 15 <212> PRT <213> Homo sapiens <400> 184 Val Lys Lys Gly Ser Trp Leu Asp Asp Phe Asn Ser Tyr Asp Arg 1 5 10 15 <210> 185 <211> 15 <212> PRT <213> Homo sapiens <400> 185 Gly Ser Trp Leu Asp Asp Phe Asn Ser Tyr Asp Arg Gln Glu Ser 1 5 10 15 <210> 186 <211> 9 <212> PRT <213> Homo sapiens <400> 186 Gly Cys Trp Leu Asp Asp Phe Asn Cys 1 5 <210> 187 <211> 15 <212> PRT <213> Homo sapiens <400> 187 Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys Tyr Ala Ser Trp 1 5 10 15 <210> 188 <211> 6 <212> PRT <213> Homo sapiens <400> 188 Trp Leu Asp Asp Phe Asn 1 5 <210> 189 <211> 5 <212> PRT <213> Homo sapiens <400> 189 Glu Gln Asp Lys Arg 1 5 <210> 190 <211> 11 <212> PRT <213> Homo sapiens <400> 190 Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr 1 5 10 <210> 191 <211> 13 <212> PRT <213> Human <400> 191 Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr 1 5 10 <210> 192 <211> 11 <212> PRT <213> Human <400> 192 Tyr Phe Cys Cys Cys Glu Gly Asn Phe Cys Asn 1 5 10 <210> 193 <211> 216 <212> PRT <213> Artificial Sequence <220> <223> Light Chain <400> 193 Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Phe Ala Gly Gly 85 90 95 Ser Tyr Tyr Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ser Thr Pro Thr Leu Thr Val Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Lys Glu Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asn Phe 130 135 140 Ser Pro Ser Gly Val Thr Val Ala Trp Lys Ala Asn Gly Thr Pro Ile 145 150 155 160 Thr Gln Gly Val Asp Thr Ser Asn Pro Thr Lys Glu Gly Asn Lys Phe 165 170 175 Met Ala Ser Ser Phe Leu His Leu Thr Ser Asp Gln Trp Arg Ser His 180 185 190 Asn Ser Phe Thr Cys Gln Val Thr His Glu Gly Asp Thr Val Glu Lys 195 200 205 Ser Leu Ser Pro Ala Glu Cys Leu 210 215 <210> 194 <211> 445 <212> PRT <213> Artificial <220> <223> Heavy Chain <400> 194 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Thr Ile Asn Pro Val Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Ala Lys Thr Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro 115 120 125 Val Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val 130 135 140 Lys Gly Tyr Phe Pro Glu Pro Val Thr Leu Thr Trp Asn Ser Gly Ser 145 150 155 160 Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu 165 170 175 Tyr Thr Leu Ser Ser Ser Val Thr Val Thr Ser Ser Thr Trp Pro Ser 180 185 190 Gln Ser Ile Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val 195 200 205 Asp Lys Lys Ile Glu Pro Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro 210 215 220 Cys Lys Cys Pro Ala Pro Asn Ala Ala Gly Gly Pro Ser Val Phe Ile 225 230 235 240 Phe Pro Pro Lys Ile Lys Asp Val Leu Met Ile Ser Leu Ser Pro Ile 245 250 255 Val Thr Cys Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln 260 265 270 Ile Ser Trp Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln 275 280 285 Thr His Arg Glu Asp Tyr Asn Ser Thr Leu Arg Val Val Ser Ala Leu 290 295 300 Pro Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys 305 310 315 320 Val Asn Asn Lys Asp Leu Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys 325 330 335 Pro Lys Gly Ser Val Arg Ala Pro Gln Val Tyr Val Leu Pro Pro Pro 340 345 350 Glu Glu Glu Met Thr Lys Lys Gln Val Thr Leu Thr Cys Met Val Thr 355 360 365 Asp Phe Met Pro Glu Asp Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys 370 375 380 Thr Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Tyr Phe Met Tyr Ser Lys Leu Arg Val Glu Lys Lys Asn Trp Val 405 410 415 Glu Arg Asn Ser Tyr Ser Cys Ser Val Val His Glu Gly Leu His Asn 420 425 430 His His Thr Thr Lys Ser Phe Ser Arg Thr Pro Gly Lys 435 440 445 BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Figure 1A graphically shows CDD866 plasma levels measured in wild-type C57BL / 6 mice treated with weekly injections of CDD866 or isotype control Ab for 8 weeks.

[0106] Figure 1B graphically shows the heart weight / tibial muscle length ratio (HW / TL) of control mice with the same type Ab (n=3) (grey bars) and experimental mice with CDD866 Ab (n=3) *p<0.05 (black bars), indicating that CDD866 does not significantly increase heart mass in adult wild-type C57BL / 6 mice.

[0107] Figure 1C is a bar graph showing % fibrosis in mice in the control group (isotype Ab, grey bars) and experimental group (CDD866 Ab [n=3], *p<0.05, black bars)). CDD866 reduced myocardial fibrosis, but the % fibrosis at baseline was significantly lower in adult wild-type C57BL / 6 mice.

[0108] Figure ID shows a representative micrograph of PAS-stained myocardium, highlighting the size of cardiomyocytes.

[0109] Figure 1E graphically illustrates the finding that CDD866 does not significantly increase cardiomyocyte size in wild-type animals. Data are presented as mean ± standard deviation. Gray = control group, isotype Ab (n = 3). Black = experimental group, CDD866 Ab (n = 3), *p < 0.05

[0110] Figure 2A graphically shows that contractile function, as measured by % fractional shortening (FS), was expectedly reduced with TAC (horizontal bars), but was preserved in CDD866-treated animals subjected to TAC (diagonal bars). SHAM + isotype Ab (n=7), black bars; SHAM + CDD866 Ab (n=7), gray bars; TAC + isotype Ab (n=10), horizontal bars; TAC + CDD866 Ab (n=10), diagonal bars. #P<0.01.

[0111] Figure 2B shows representative echocardiographic images 11 weeks after SHAM or TAC surgery, demonstrating preserved contractile function in TAC animals treated with CDD866.

[0112] Figure 2C graphically shows the lung weight / tibialis muscle length ratio (LW / TL) of mice in different treatment groups. SHAM + isotype Ab (n=7), black bars; SHAM + CDD866 Ab (n=7), gray bars; TAC + isotype Ab (n=10), horizontal bars; TAC + CDD866 Ab (n=10), diagonal bars. *p<0.01. There was a trend toward decreased lung weight in CDD866-treated animals, indicating less lung congestion (a surrogate for heart failure in the mouse model).

[0113] Figure 2D graphically shows a significant reduction in the primary endpoint (survival or %FS < 20%) with CDD866 treatment.

[0114] FIG3A graphically shows plasma CDD866 levels for various treatment groups: TAC+isotype; TAC+CDD866; Sham+isotype; and Sham+CDD866.

[0115] Figure 3B is a bar graph showing that cardiac follistatin-like 3 (FSTL3) expression increases with TAC, indicating increased cardiac ActRII-A / B signaling in this cardiac injury model. CDD866 treatment reduces cardiac FSTL3 expression, indicating that it effectively blocks TAC-induced ActRII-A / B signaling in the heart. Black = SHAM + isotype Ab (n = 7). Gray = SHAM + CDD866 Ab (n = 7). Horizontal bars = TAC + isotype Ab (n = 10). Diagonal bars = TAC + CDD866 Ab (n = 10). *p < 0.05. #p < 0.01.

[0116] Figure 3C graphically shows that the relative mRNA expression of pathological cardiac hypertrophy genes decreased with CDD866 treatment. ANP (atrial natriuretic peptide); BNP (brain natriuretic peptide); aMHC (alpha myosin heavy chain); bMHC (beta myosin heavy chain). Black = SHAM + isotype Ab (n = 7). Gray = SHAM + CDD866 Ab (n = 7). Horizontal bars = TAC + isotype Ab (n = 10). Diagonal bars = TAC + CDD866 Ab (n = 10). *p < 0.05. #p < 0.01.

[0117] FIG3D is a bar graph illustrating that treatment with CDD866 reduces relative mRNA expression of pathological cardiac fibrosis genes in TAC-induced heart failure. COL1 (collagen type 1); CTGF (connective tissue growth factor). Black = SHAM + isotype Ab (n=7). Gray = SHAM + CDD866 Ab (n=7). Horizontal bars = TAC + isotype Ab (n=10). Diagonal bars = TAC + CDD866 Ab (n=10). *p<0.05. #p<0.01.

[0118] FIG4A graphically shows CDD866 plasma levels measured in mice that developed contractile function after TAC followed by weekly treatment with CDD866 injections for 8 weeks.

[0119] Figure 4B is a bar graph showing the relative mRNA expression levels of FSTL3 (follistatin-like 3), activin A, MSTN (myostatin), ACVR2A (activin A receptor type 2A), and ACVR2B (activin A receptor type 2B). The graph shows that treatment with CDD866 can reduce cardiac FSTL3 expression, indicating that CDD866 can effectively block TAC-induced ActRII-A / B signaling in the heart.

[0120] Figure 4C is a graph of % fractional shortening plotted against time (in weeks), showing that CDD866 reversed systolic dysfunction in TAC-induced heart failure in a progressively improving manner as early as 1 week after treatment.

[0121] Figure 4D graphically shows that CDD866 also reduced the lung weight to tibialis muscle length ratio, a surrogate marker for heart failure in a mouse model. Gray = TAC + isotype Ab. Black = TAC + CDD866 Ab. *p < 0.05. #p < 0.01. LW / TL (lung weight / tibialis muscle length ratio).

[0122] Figure 5A is a graph plotting wall thickness versus weeks after TAC, showing a gradual increase in wall thickness with CDD866 treatment. The arrow indicates the onset of CDD866.

[0123] Figure 5B shows serial echo images of ventricular sections during treatment, demonstrating differences in cardiac growth between isotype- and CDD866-treated animals. Cdd866-mediated cardiac growth prevents eccentric remodeling associated with progressive contractile dysfunction.

[0124] Figure 5C is a graph showing the heart weight / tibialis muscle length ratio (HW / TL) of mice treated with TAC+isotype Ab (grey bars) and TAC+CDD866 Ab (black bars), demonstrating that CDD866 increases heart mass in the TAC model. *p<0.05. #p<0.01.

[0125] Figure 5D shows micrographs of PAS-stained myocardium highlighting cardiomyocyte size in TAC+isotype Ab and TAC+CDD866Ab treated mice.

[0126] Figure 5E is a graph showing cross-sectional area of ​​cardiomyocytes from mice treated with TAC+isotype Ab (grey) and TAC+CDD866Ab (black), demonstrating that CDD866 increases cardiomyocyte growth in TAC. *p<0.05. #p<0.01.

[0127] FIG6A is a graph showing that mRNA expression of genes associated with pathological hypertrophy decreased with CDD866 treatment. ANP (atrial natriuretic peptide); BNP (brain natriuretic peptide); αMHC (α-myosin heavy chain); βMHC (β-myosin heavy chain). TAC + isotype Ab (grey); TAC + CDD866 Ab (black) are shown. *p<0.05. #p<0.01.

[0128] Figure 6B plots fractional shortening, wall thickness, and body weight of mice over time (in weeks). Arrow = timing of single dose; dashed line = expected trajectory without CDD866 treatment. The figure shows that the effects of CDD866 on cardiac growth and body weight occur rapidly, are transient, and reversible. The effects of a single dose of CDD866 also occur within a 1-2 week timeframe and persist for at least 6 weeks.

[0129] FIG6C shows a photomicrograph of Masson's trichrome-stained myocardium (blue = fibrosis; red = muscle), demonstrating reduced cardiac fibrosis in TAC'd mice treated with CDD866.

[0130] Figure 6D is a bar graph showing % fibrosis using TAC + isotype Ab (grey) and TAC + CDD866 Ab (black), demonstrating a trend toward reduced myocardial fibrosis with CDD866 treatment. *p<0.05. #p<0.01.

[0131] Figure 7A is a Western blot of gastrocnemius muscle samples probed with p-SMAD3 and GAPDH antibodies. Samples were collected from T57BL / 6 mice with confirmed cardiac dysfunction after TAC and subsequently treated with CDD866 (TT-2 to TT-10) or isotype control Abs (TT-11 to TT-20) for 8 weeks. This figure generally demonstrates that CDD866 reduces ActRII-A / B signaling in skeletal muscle in a TAC-mediated mouse model of heart failure. MSTN stimulation in C2C12 cells was used as a positive control for the assay.

[0132] Figure 7B is a graph showing the % change in mouse weight from baseline measured relative to weeks post-treatment. Diamond data points (red) represent TAC + isotype Ab. Square data points (blue) represent TAC + CDD866 Ab. *p<0.05. #p<0.01. CDD866 resulted in a gradual increase in overall body weight, likely through increased muscle mass.

[0133] Figure 7C graphically shows the % change in muscle mass relative to control for each of the following skeletal muscle groups: EDL (extensor digitorum longus), gastrocnemius, and tibialis. Red indicates TAC + isotype Ab. Blue indicates TAC + CDD866 Ab. *p<0.05. #p<0.01. CDD866 increased the mass of all three skeletal muscle groups overall.

[0134] Figure 7D is a graph showing % fiber distribution relative to serial tissue sections, demonstrating that CDD866 increases skeletal muscle cell size. Red indicates TAC + isotype Ab treatment. Blue indicates TAC + CDD866 Ab treatment. *p<0.05. #p<0.01.

[0135] Figure 7E shows four graphs showing the % fiber distribution relative to serial tissue sections, demonstrating that CDD866 induces multiple fiber type switching in skeletal muscle. Red indicates TAC+isotype Ab treatment. Blue indicates TAC+CDD866 Ab treatment.

[0136] FIG8A is a graph showing changes in % fractional shortening (FS) over time in mice harboring a missense mutation (F764L) in the αMHC gene (a mouse model of dilated cardiomyopathy). Twelve weeks of CDD866 treatment resulted in a trend toward a modest increase in contractile function. Gray = isotype Ab. Black = CDD866 Ab. *p<0.05. #p<0.01.

[0137] Figure 8B is a bar graph showing the relative mRNA expression levels of various genes associated with the ActRII pathway in cardiac tissue from mice treated with CDD866 (black) or isotype Ab (grey). There was a trend toward decreased cardiac FSTL3 expression with CDD866, suggesting inhibition of ActRII receptor signaling in the heart. *p<0.05. #p<0.01.

[0138] FIG8C is a bar graph showing relative mRNA expression levels of genes associated with pathological hypertrophy in mice treated with isotype Ab or CDD866 Ab. *p<0.05. #p<0.01. Gray = isotype Ab. Black = CDD866 Ab. *p<0.05. #p<0.01. ANP (atrial natriuretic peptide); BNP (brain natriuretic peptide); αMHC (α-myosin heavy chain); βMHC (β-myosin heavy chain). There were no significant differences in the pathological hypertrophy gene expression profiles.

[0139] definition

[0140] In order to more readily understand the present disclosure, certain terms are first defined. Additional definitions are set forth throughout the detailed description.

[0141] The term "comprising" means "including", eg, a composition "comprising" X may consist solely of X or may include something additional, eg, X+Y.

[0142] The term "about" in connection with a value x means, for example, x ± 10%.

[0143] The following exemplifies possible preclinical treatment protocols to evaluate the potential effects of treatment with an ActRII binding molecule, more preferably an antagonist antibody to ActRII (eg, bimagrumab).

[0144] Treatment is exemplified using mice subjected to transverse aortic constriction (TAC), a common experimental model of pressure overload-induced cardiac hypertrophy and heart failure. Technicians know how to establish suitable experimental or dosing protocols for other species, particularly humans. For studies in primates, anti-ActRII antibodies (e.g., bimagrumab) can be administered to male and female macaques once a week by intravenous injection for up to 3 months. 32 macaques (16 / sex) can be assigned to one of four treatment groups (3 to 5 animals / sex / group) and can be administered vehicle or ActRIIB antibody (e.g., BYM338) at 10, 30, or 100 mg / kg intravenously once a week for 13 weeks (a total of 14 doses; the dose should be selected based on the symptoms of the heart disease).

[0145] The terms "ActRIIA" and "ActRIIB" refer to activin receptors. Activins signal through a heterodimeric complex of receptor serine kinases, which include at least two type I (I and IB) and two type II (IIA and IIB, also known as ACVR2A and ACVR2B) receptors. These receptors are all transmembrane proteins consisting of a ligand-binding extracellular domain (which has a cysteine-rich region), a transmembrane domain, and a cytoplasmic domain with predicted serine / threonine specificity. Type I receptors are essential for signal transduction, while type II receptors are required for ligand binding and expression / recruitment of type I receptors. Type I and type II receptors form a stable complex after ligand binding, resulting in phosphorylation of type I receptors by type II receptors. Activin receptor IIB (ActRIIB) is a receptor for myostatin. Activin receptor IIA (ActRIIA) is also a receptor for myostatin. The term ActRIIB or Act IIB receptor refers to human ActRIIB as defined in SEQ ID NO: 181 (AAC64515.1, GI: 3769443). Research grade polyclonal and monoclonal anti-ActRIIB antibodies are known in the art, such as those produced by R&D Of course, antibodies can be raised against ActRIIB from other species and used to treat pathological conditions in those species.

[0146] The term "immune response" refers to the action of, for example, lymphocytes, antigen-presenting cells, phagocytes, granulocytes, and soluble macromolecules (e.g., antibodies, cytokines, and complement) produced by these cells or the liver that results in the selective damage, destruction, or elimination from the body of invading pathogens, cells or tissues infected by pathogens, cancer cells, or, in the case of autoimmunity or pathological inflammation, normal human cells or tissues.

[0147] "Signaling activity" refers to a biochemical cause-and-effect relationship, typically initiated by protein-protein interactions (such as the binding of a growth factor to a receptor), resulting in the transmission of a signal from one part of a cell to another. Typically, the transmission involves specific phosphorylation of one or more tyrosine, serine, or threonine residues on one or more proteins in a series of reactions, leading to signal transduction. The penultimate process typically involves nuclear events, resulting in changes in gene expression.

[0148] The term "antibody" as used herein includes intact antibodies and any antigen-binding fragments (i.e., "antigen-binding portions") or single chains thereof. Naturally occurring "antibodies" are glycoproteins comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as V H) and a heavy chain constant region. The heavy chain constant region is composed of three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as V L ) and the light chain constant region. The light chain constant region consists of a domain C L Composition. H and V L The V domains can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), and more conserved regions, called framework regions (FRs), interspersed with the CDRs. H and V L It is composed of three CDRs and four FRs, which are arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0149] As used herein, the term "antigen-binding portion" of an antibody (or simply "antigen portion") refers to the full length of an antibody, or one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., a portion of ActRIIB). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody include: a Fab fragment, which is a fragment of a V-binding region of a Fab protein. L 、V H 、C L and CH1 domains; F(ab)2 fragment, which is a bivalent fragment comprising two Fab fragments, each of which binds to the same antigen and is linked by a disulfide bond in the hinge region; and Fd fragment, which consists of V H and CH1 domains; Fv fragment, the Fv fragment consists of the V L and V H Domain composition; dAb fragment (Ward et al., 1989 Nature 341:544-546), the dAb fragment consists of V H domain composition; and isolated complementarity determining regions (CDRs).

[0150] In addition, although the two domains V L and V Hare encoded by separate genes, but they can be joined using recombinant methods by a synthetic linker that enables them to be made into a single protein chain in which V L and V H The regions are paired to form monovalent molecules (called single-chain Fv (scFv); see, e.g., Bird et al., 1988 Science 242:423-426; and Huston et al., 1988 Proc. Natl. Acad. Sci. 85:5879-5883). Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding region" of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for use in the same manner as intact antibodies.

[0151] As used herein, an "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigenic specificities (e.g., an isolated antibody that specifically binds ActRIIB is substantially free of antibodies that specifically bind an antigen other than ActRIIB). However, an isolated antibody that specifically binds ActRIIB may have cross-reactivity with other antigens, such as ActRIIB molecules from other species. Furthermore, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0152] The term "cross-blocking" refers to the ability of an antibody or other binding agent to interfere with the binding of other antibodies or binding agents to ActRIIB, particularly the ligand binding domain, in a standard competitive binding assay.

[0153] The term "monoclonal antibody" or "monoclonal antibody composition" as used herein refers to a preparation of antibody molecules of single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.

[0154] As used herein, the term "human (class) antibody " is intended to include such antibodies, and the antibody has a variable region, in which framework region and CDR region are all derived from sequences in human origin. In addition, if the antibody contains a constant region, the constant region is also derived from this type of human sequence, such as human germline sequence, or a mutant form of human germline sequence, or an antibody containing a consensus framework sequence derived from human framework sequence analysis, such as described in Knappik et al. (2000.J Mol Biol 296,57-86). Human antibodies of the present disclosure may include amino acid residues not encoded by human sequences (such as mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, as used herein, the term "human (class) antibody " is not intended to include such antibodies, in which the CDR sequences derived from the germline of another mammalian species (such as a mouse) have been transplanted onto human framework sequences.

[0155] The term "human monoclonal antibody" refers to an antibody displaying a single binding specificity having a variable region in which both the framework and CDR regions are derived from human sequences. In one embodiment, the human monoclonal antibody is produced by a hybridoma comprising a B cell obtained from a transgenic non-human animal, such as a transgenic mouse, having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell.

[0156] As used herein, the term "recombinant human (class) antibody" includes all human antibodies prepared, expressed, produced or isolated by recombinant methods, such as antibodies isolated from animals (e.g., mice) that are transgenic or transchromosomal for human immunoglobulin genes or hybridomas prepared therefrom, antibodies isolated from host cells transformed to express human antibodies (e.g., antibodies from transfectomas), antibodies isolated from recombinant, combinatorial human antibody libraries, and antibodies prepared, expressed, produced or isolated by any other method involving splicing all or part of the sequence of human immunoglobulin genes to other DNA sequences. Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies can be subjected to in vitro mutagenesis (or, when using animals that are transgenic for human Ig sequences, somatic mutagenesis in the receptor), whereby the V H Area and V L The amino acid sequence of the region is such a sequence that, although derived from human germline V H Sequence and V L Sequence and human germline V H Sequence and V L Sequence-related, but may not naturally occur in the human antibody germline repertoire.

[0157] As used herein, "isotype" refers to the antibody class provided by the heavy chain constant region genes (eg, IgM, IgE, IgG, such as IgGl or IgG2).

[0158] The phrases "an antibody that recognizes an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody that specifically binds to an antigen."

[0159] As used herein, an antibody that "specifically binds to an ActRIIB polypeptide" means an antibody that binds to an ActRIIB polypeptide with a K of about 100 nM or less, about 10 nM or less, about 1 nM or less, or about 1 nM or less. D An antibody that "cross-reacts with an antigen other than ActRIIB" means an antibody that binds to a human ActRIIB polypeptide at a concentration of about 10 × 10 -9 M or lower, about 5×10 -9 M or lower, or about 2×10 -9 M or lower K D An antibody that "does not cross-react with a specific antigen" means an antibody that binds to an antigen at a rate of about 1.5 × 10 -8 M or higher K D , or about 5-10×10 -8 M or about 1×10 -7 M or higher K D In certain embodiments, such antibodies that do not cross-react with the antigen exhibit substantially undetectable binding to these proteins in standard binding assays. Biosensor systems such as Equilibrium titration of the system or solution to determine K D .

[0160] As used herein, the term "antagonist antibody" refers to an antibody that inhibits ActRIIB-induced signaling activity in the presence of myostatin or other ActRIIB ligands (such as activin or GDF-11), and / or refers to an antibody that inhibits ActRIIA-induced signaling activity in the presence of myostatin or other ActRIIA ligands (such as activin or GDF-11). Examples of assays for detecting this include inhibition of myostatin-induced signaling (e.g., by Smad-dependent reporter gene assay), inhibition of myostatin-induced Smad phosphorylation (P-Smad ELISA), and inhibition of myostatin-induced inhibition of skeletal muscle cell differentiation (e.g., by creatine kinase assay).

[0161] In some embodiments, the antibody inhibits myostatin-induced signaling as measured in a Smad-dependent reporter gene assay, IC 50About 10 nM or less, about 1 nM or less, or about 100 pM or less.

[0162] As used herein, an antibody with "no agonistic activity" is intended to mean an antibody that does not significantly increase ActRIIB-mediated signaling activity in the absence of myostatin in a cell-based assay, such as inhibition of myostatin-induced signaling (e.g., by a Smad-dependent reporter gene assay), inhibition of myostatin-induced Smad phosphorylation (P-Smad ELISA), and inhibition of myostatin-induced inhibition of skeletal muscle cell differentiation (e.g., by a creatine kinase assay).

[0163] As used herein, the term "K assoc ” or “K a " is intended to represent the association rate of a particular antibody-antigen interaction, while the term "K" as used herein dis ” or “K d " is intended to represent the off-rate of a particular antibody-antigen interaction. As used herein, the term "K D " is intended to represent the dissociation constant, which is derived from K d With K a The ratio (K d / K a ) and expressed as molar concentration (M). The K of an antibody can be determined using well-established methods in the art. D Used to determine the K value of the antibody D The method is to use surface plasmon resonance, such as Biosensor systems, or solution equilibrium titration (SET) (see Friguet B et al., (1985) J. Immunol Methods; 77(2):305-319, and Hanel C et al., (2005) Anal Biochem; 339(1):182-184).

[0164] As used herein, the term "affinity" refers to the strength of the interaction between an antigen and an antibody at a single antigenic site. Within each antigenic site, the variable region of the antibody "arm" interacts with the antigen at many sites through weak non-covalent forces; the more interactions there are, the stronger the affinity.

[0165] As used herein, the term "avidity" refers to an informative measure of the overall stability or strength of an antibody-antigen complex. It is governed by three main factors: antibody epitope affinity; the valence of both the antigen and the antibody; and the structural arrangement of the interacting parts. Ultimately, these factors define the specificity of an antibody, i.e., the likelihood that a particular antibody will bind to a precise antigen epitope.

[0166] As used herein, the term "ADCC" or "antibody-dependent cellular cytotoxicity" activity refers to human B cell depletion activity. ADCC activity can be measured by a human B cell depletion assay known in the art.

[0167] In order to obtain higher affinity probes, dimer conjugates (two antibody protein molecules coupled to a FACS marker) can be constructed to make it easier to detect low affinity interactions (such as interactions with germline antibodies) by FACS. In addition, another method for increasing antigen binding affinity involves producing dimers, trimers, or multimers of any construct of the anti-ActRIIB antibodies described herein. Such multimers can be produced by covalent bonding between the individual modules, for example, by simulating natural C-to-N-terminal binding or by simulating antibody dimers held together by the constant region of the antibody. The bonds engineered into the Fc / Fc interface can be covalent or non-covalent. In addition, dimerization or multimerization of partners other than Fc can be used in ActRIIB hybrids to create such higher order structures. For example, a multimerization domain can be used, such as the trimerization domain described in WO2004 / 039841 or the pentamerization domain described in WO98 / 18943.

[0168] As used herein, the term "selectivity" of an antibody refers to the ability of the antibody to bind to a particular target polypeptide but not to closely related polypeptides.

[0169] As used herein, the term "high affinity" of an antibody refers to the K of the antibody for the target antigen. D As used herein, the term "subject" includes any human or non-human animal.

[0170] The term "non-human animals" includes all vertebrates, eg, mammals and non-mammals, such as non-human primates, sheep, dogs, cats, mice, horses, cows, chickens, amphibians, reptiles, etc.

[0171] As used herein, the term "optimized" refers to a nucleotide sequence that has been altered to encode an amino acid sequence using codons that are preferred in production cells or organisms, typically eukaryotic cells, such as cells of Pichia, Trichoderma, Chinese hamster ovary cells (CHO), or human cells. The optimized nucleotide sequence is engineered to retain, completely or as much as possible, the amino acid sequence originally encoded by the starting nucleotide sequence, which is also referred to as the "parent" sequence. The optimized sequences herein have been engineered to have codons that are preferred in CHO mammalian cells, but optimized expression of these sequences in other eukaryotic cells is also contemplated herein. The amino acid sequence encoded by the optimized nucleotide sequence is also referred to as optimized.

[0172] Detailed description of the present disclosure

[0173] Antibodies directed against ActRII receptors, such as bimagrumab, have been found to reduce signaling through these receptors and lead to the prevention and / or treatment of heart disease.

[0174] Therefore, in one aspect, the present disclosure provides a composition comprising an ActRIIA or ActRIIB binding molecule (e.g., bimagrumab), or a functional protein comprising an antigen binding portion of the antibody. The binding molecule may be an ActRIIB binding molecule, such as human ActRIIB. The polypeptide sequence of human ActRIIB is shown in SEQ ID NO: 181 (AAC64515.1, GI: 3769443). In one embodiment, the antibody or functional protein is from a mammal, having an origin such as human or camelid. Therefore, the antibody contained in the disclosed composition may be a chimeric antibody, a human antibody, or a humanized antibody. In a specific embodiment, the anti-ActRIIB antibody contained in the disclosed composition is characterized by having an antigen binding region that is specific for the target protein ActRIIB and binds to ActRIIB or a fragment of ActRIIB.

[0175] In one embodiment, the antibody contained in the disclosed composition is an ActRII antagonist that has no agonistic activity or has low agonistic activity. In another embodiment, the antibody or functional fragment contained in the disclosed composition binds to the target protein ActRII and reduces the binding of myostatin to ActRII to a basal level. In another aspect of this embodiment, the antibody or functional fragment contained in the disclosed composition completely prevents the binding of myostatin to ActRII. In another embodiment, the antibody or functional fragment contained in the disclosed composition inhibits Smad activation. In another embodiment, the antibody or functional fragment contained in the disclosed composition inhibits type IIB activin receptor-mediated myostatin-induced inhibition of skeletal differentiation through a Smad-dependent pathway.

[0176] Binding can be determined by one or more assays that can be used to measure the antagonistic or agonistic activity of the antibody. Preferably, the assay measures at least one of the effects of the antibody on ActRIIB, including: inhibiting myostatin binding to ActRIIB (by ELISA); inhibiting myostatin-induced signaling (e.g., by Smad-dependent reporter gene assay), inhibiting myostatin-induced Smad phosphorylation (P-Smad ELISA), and inhibiting myostatin-induced inhibition of skeletal muscle cell differentiation (e.g., by creatine kinase assay).

[0177] In one embodiment, the present disclosure provides a composition comprising an antibody that specifically binds to the myostatin binding region (i.e., the ligand binding domain) of ActRIIB. The ligand binding domain consists of amino acids 19-134 of SEQ ID NO: 181 and has been designated herein as SEQ ID NO: 182. The ligand binding domain comprises several epitopes described below.

[0178] In one embodiment, the antibodies included in the disclosed compositions have a K of about 100 nM or less, about 10 nM or less, about 1 nM or less, or about 1 nM or less. D Binding to ActRIIB. Preferably, the antibodies included in the disclosed compositions bind to ActRIIB with an affinity of 100 pM or less (i.e., about 100 pM, about 50 pM, about 10 pM, about 2 pM, about 1 pM, or less). In one embodiment, the antibodies included in the disclosed compositions bind to ActRIIB with an affinity of between about 1 pM and about 10 pM.

[0179] In one embodiment, the antibodies included in the disclosed compositions do not cross-react with ActRIIB-related proteins, in particular, do not cross-react with human ActRIIA (NP_001607.1, GI: 4501897). In another embodiment, the antibodies included in the disclosed compositions cross-react with ActRIIA and bind to ActRIIB with equivalent affinity, or bind to ActRIIB with an affinity that is about 1-fold, 2-fold, 3-fold, 4-fold, or 5-fold, more preferably about 10-fold, still more preferably about 20-fold, 30-fold, 40-fold, or 50-fold, and still more preferably about 100-fold greater than their affinity for ActRIIA.

[0180] In one embodiment, the antibodies comprised in the disclosed compositions bind to ActRIIA with an affinity of 100 pM or greater (ie, about 250 pM, about 500 pM, about 1 nM, about 5 nM or greater).

[0181] In one embodiment, the antibodies comprised in the disclosed compositions are of the IgG2 isotype.

[0182] In another embodiment, the antibody included in the disclosed composition is of the IgG1 isotype. In another embodiment, the antibody included in the disclosed composition is of the IgG1 isotype and has altered effector function by mutation of the Fc region. The altered effector function can be reduced ADCC activity and CDC activity. In one embodiment, the altered effector function is silenced ADCC activity and CDC activity.

[0183] In another related embodiment, the antibody comprised in the disclosed composition is a fully human or humanized IgG1 antibody that has no antibody-dependent cellular cytotoxicity (ADCC) activity or CDC activity and binds to the ActRIIB region consisting of amino acids 19-134 of SEQ ID NO:181.

[0184] In another related embodiment, the antibody comprised in the disclosed composition is a fully human or humanized IgG1 antibody having reduced antibody-dependent cellular cytotoxicity (ADCC) activity or CDC activity and binding to the ActRIIB region consisting of amino acids 19-134 of SEQ ID NO:181.

[0185] The present disclosure also relates to compositions comprising human anti-ActRIIB antibodies or humanized anti-ActRIIB antibodies for use in preventing and / or treating heart diseases as described above.

[0186] In certain embodiments, the antibodies included in the disclosed compositions are derived from specific heavy and light chain sequences and / or contain specific structural features, such as CDR regions comprising specific amino acid sequences. The present disclosure provides isolated ActRIIB antibodies, methods for preparing the antibodies, immunoconjugates and multivalent or multispecific molecules comprising the antibodies, and pharmaceutical compositions containing the antibodies, immunoconjugates, or bispecific molecules.

[0187] In alternative embodiments, the present disclosure relates to the following aspects:

[0188] 1. An ActRII receptor antagonist for use in treating and / or preventing heart failure, including heart failure associated with or caused by valvular heart disease, hypertension, coronary artery disease, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, and other forms of inherited or idiopathic cardiomyopathy.

[0189] 2. The ActRII receptor antagonist for use according to aspect 1, wherein the ActRII antagonist is administered to a patient in need thereof at a dose of about 3-10 mg / kg.

[0190] 3. The ActRII receptor antagonist for use according to aspect 2, wherein the myostatin antagonist is administered at a dose of about 3 mg / kg body weight or about 10 mg / kg body weight.

[0191] Alternatively, the ActRII receptor antagonist is administered at a dose of about 3, 4, 5, 6, 7, 8, 9, or about 10 mg / kg body weight.

[0192] 4. The ActRII receptor antagonist for use according to aspects 1-3, wherein the ActRII receptor antagonist is administered intravenously or subcutaneously.

[0193] 5. An ActRII receptor antagonist for use according to any one of aspects 1 to 4, wherein the ActRII receptor antagonist is administered once every four weeks.

[0194] Alternatively, the ActRII receptor antagonist may be administered once every 8 weeks.

[0195] 6. An ActRII receptor antagonist for use according to any one of aspects 1 to 5, wherein the ActRII receptor antagonist is administered for at least 3 months.

[0196] 7. An ActRII receptor antagonist for use according to any one of aspects 1 to 6, wherein the ActRII receptor antagonist is administered for up to 12 months.

[0197] Preferably, the ActRII receptor antagonist is administered for at least or at most 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months.

[0198] 8. A method for treating and / or preventing heart failure, comprising administering an effective amount of an ActRII receptor antagonist to a subject having heart failure or at risk of developing heart failure.

[0199] In many cases, heart failure can be caused by or associated with conditions such as valvular heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, exposure to toxic and infectious agents, and other forms of inherited or idiopathic cardiomyopathy. Patients at risk for developing heart failure may have one or more of these conditions.

[0200] 9. A method of treating structural and / or functional cardiac abnormalities associated with a condition selected from the group consisting of valvular heart disease, hypertension, coronary artery disease, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic and infectious agents, and other forms of inherited or idiopathic cardiomyopathy, the method comprising administering to a subject having the structural and / or functional cardiac abnormalities associated with the condition an effective amount of an ActRII receptor antagonist.

[0201] 10. The method of aspect 8 or 9, comprising administering the ActRII receptor antagonist to a patient in need thereof at a dose of about 3-10 mg / kg.

[0202] 11. The method of aspect 8 or 9, comprising administering the ActRII receptor antagonist to a patient in need thereof at a dose of about 3 or about 10 mg / kg body weight.

[0203] 12. The method of aspect 8 or 9, comprising administering the ActRII receptor antagonist intravenously or subcutaneously.

[0204] 13. The method of any one of aspects 8 to 10, comprising administering the ActRII receptor antagonist once every four weeks.

[0205] 14. The method of any one of aspects 8 to 13, comprising administering the ActRII receptor antagonist for at least 3 months.

[0206] 15. The method of aspect 14, comprising administering the ActRII receptor antagonist for up to 12 months.

[0207] 16. An ActRII receptor antagonist for the method or use according to any one of aspects 1-15, wherein the ActRII receptor antagonist is an anti-ActRII receptor antibody.

[0208] 17. An ActRII receptor antagonist for the method or use according to any one of aspects 1-16, wherein the anti-ActRII receptor antibody is bimagrumab.

[0209] 18. An ActRII receptor antagonist for use according to aspect 17, wherein the ActRII receptor antagonist is an anti-ActRII antibody that binds to an epitope of ActRIIB consisting of amino acids 19-134 of SEQ ID NO: 181 (SEQ ID NO: 182).

[0210] 19. The method or ActRII receptor antagonist for use according to any one of aspects 16-18, wherein the anti-ActRII antibody binds to an epitope of ActRIIB comprising or consisting of:

[0211] (a) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188);

[0212] (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC-SEQ ID NO: 186);

[0213] (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190);

[0214] (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR—SEQ ID NO: 189);

[0215] (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW-SEQ ID NO: 187);

[0216] (f) amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191);

[0217] (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN - SEQ ID NO: 192); or

[0218] (h) Amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR).

[0219] 20. An ActRII receptor antagonist for use according to any one of aspects 16 to 19, wherein the anti-ActRIIB antibody is selected from the group consisting of:

[0220] a) an anti-ActRIIB antibody, wherein the anti-ActRIIB antibody binds to an epitope of ActRIIB comprising:

[0221] (a) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188);

[0222] (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC-SEQ ID NO: 186);

[0223] (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190);

[0224] (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR—SEQ ID NO: 189);

[0225] (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW-SEQ ID NO: 187);

[0226] (f) amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191);

[0227] (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN - SEQ ID NO: 192); or

[0228] (h) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR);

[0229] and b) an ActRIIB antagonist antibody that binds to an epitope of ActRIIB comprising amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO: 188);

[0230] (b) amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC-SEQ ID NO: 186);

[0231] (c) amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO: 190);

[0232] (d) amino acids 52-56 of SEQ ID NO: 181 (EQDKR—SEQ ID NO: 189);

[0233] (e) amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW-SEQ ID NO: 187);

[0234] (f) amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191);

[0235] (g) amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN - SEQ ID NO: 192); or

[0236] (h) amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR), wherein the K D It is about 2pM.

[0237] 21. An ActRII receptor antagonist for the method or use of any one of aspects 16-20, wherein the antibody binds to ActRIIB with an affinity that is 10 times greater than its affinity for ActRIIA.

[0238] 22. The method or ActRII receptor antagonist for use according to any one of aspects 16-21, wherein the antibody comprises: a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14; a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28; a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: 42; a light chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 43 to SEQ ID NO: 56; a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 57 to SEQ ID NO: 70; and a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 71 to SEQ ID NO: The amino acid sequence of the group consisting of NO:84.

[0239] 23. An ActRII receptor antagonist for the method or use according to any one of aspects 16 to 22, wherein the antibody comprises:

[0240] (a) the heavy chain variable region CDR1 set forth in SEQ ID NO: 1; the heavy chain variable region CDR2 set forth in SEQ ID NO: 15; the heavy chain variable region CDR3 set forth in SEQ ID NO: 29; the light chain variable region CDR1 set forth in SEQ ID NO: 43; the light chain variable region CDR2 set forth in SEQ ID NO: 57; and the light chain variable region CDR3 set forth in SEQ ID NO: 71,

[0241] (b) the heavy chain variable region CDR1 set forth in SEQ ID NO: 2; the heavy chain variable region CDR2 set forth in SEQ ID NO: 16; the heavy chain variable region CDR3 set forth in SEQ ID NO: 30; the light chain variable region CDR1 set forth in SEQ ID NO: 44; the light chain variable region CDR2 set forth in SEQ ID NO: 58; and the light chain variable region CDR3 set forth in SEQ ID NO: 72,

[0242] (c) the heavy chain variable region CDR1 set forth in SEQ ID NO: 3; the heavy chain variable region CDR2 set forth in SEQ ID NO: 17; the heavy chain variable region CDR3 set forth in SEQ ID NO: 31; the light chain variable region CDR1 set forth in SEQ ID NO: 45; the light chain variable region CDR2 set forth in SEQ ID NO: 59; and the light chain variable region CDR3 set forth in SEQ ID NO: 73,

[0243] (d) the heavy chain variable region CDR1 set forth in SEQ ID NO:4; the heavy chain variable region CDR2 set forth in SEQ ID NO:18; the heavy chain variable region CDR3 set forth in SEQ ID NO:32; the light chain variable region CDR1 set forth in SEQ ID NO:46; the light chain variable region CDR2 set forth in SEQ ID NO:60; and the light chain variable region CDR3 set forth in SEQ ID NO:74,

[0244] (e) the heavy chain variable region CDR1 set forth in SEQ ID NO:5; the heavy chain variable region CDR2 set forth in SEQ ID NO:19; the heavy chain variable region CDR3 set forth in SEQ ID NO:33; the light chain variable region CDR1 set forth in SEQ ID NO:47; the light chain variable region CDR2 set forth in SEQ ID NO:61; and the light chain variable region CDR3 set forth in SEQ ID NO:75,

[0245] (f) the heavy chain variable region CDR1 set forth in SEQ ID NO:6; the heavy chain variable region CDR2 set forth in SEQ ID NO:20; the heavy chain variable region CDR3 set forth in SEQ ID NO:34; the light chain variable region CDR1 set forth in SEQ ID NO:48; the light chain variable region CDR2 set forth in SEQ ID NO:62; and the light chain variable region CDR3 set forth in SEQ ID NO:76,

[0246] (g) the heavy chain variable region CDR1 set forth in SEQ ID NO:7; the heavy chain variable region CDR2 set forth in SEQ ID NO:21; the heavy chain variable region CDR3 set forth in SEQ ID NO:35; the light chain variable region CDR1 set forth in SEQ ID NO:49; the light chain variable region CDR2 set forth in SEQ ID NO:63; and the light chain variable region CDR3 set forth in SEQ ID NO:77,

[0247] (h) the heavy chain variable region CDR1 set forth in SEQ ID NO:8; the heavy chain variable region CDR2 set forth in SEQ ID NO:22; the heavy chain variable region CDR3 set forth in SEQ ID NO:36; the light chain variable region CDR1 set forth in SEQ ID NO:50; the light chain variable region CDR2 set forth in SEQ ID NO:64; and the light chain variable region CDR3 set forth in SEQ ID NO:78,

[0248] (i) the heavy chain variable region CDR1 set forth in SEQ ID NO:9; the heavy chain variable region CDR2 set forth in SEQ ID NO:23; the heavy chain variable region CDR3 set forth in SEQ ID NO:37; the light chain variable region CDR1 set forth in SEQ ID NO:51; the light chain variable region CDR2 set forth in SEQ ID NO:65; and the light chain variable region CDR3 set forth in SEQ ID NO:79,

[0249] (j) the heavy chain variable region CDR1 set forth in SEQ ID NO:10; the heavy chain variable region CDR2 set forth in SEQ ID NO:24; the heavy chain variable region CDR3 set forth in SEQ ID NO:38; the light chain variable region CDR1 set forth in SEQ ID NO:52; the light chain variable region CDR2 set forth in SEQ ID NO:66; and the light chain variable region CDR3 set forth in SEQ ID NO:80,

[0250] (k) the heavy chain variable region CDR1 set forth in SEQ ID NO:11; the heavy chain variable region CDR2 set forth in SEQ ID NO:25; the heavy chain variable region CDR3 set forth in SEQ ID NO:39; the light chain variable region CDR1 set forth in SEQ ID NO:53; the light chain variable region CDR2 set forth in SEQ ID NO:67; and the light chain variable region CDR3 set forth in SEQ ID NO:81,

[0251] (1) the heavy chain variable region CDR1 set forth in SEQ ID NO:12; the heavy chain variable region CDR2 set forth in SEQ ID NO:26; the heavy chain variable region CDR3 set forth in SEQ ID NO:40; the light chain variable region CDR1 set forth in SEQ ID NO:54; the light chain variable region CDR2 set forth in SEQ ID NO:68; and the light chain variable region CDR3 set forth in SEQ ID NO:82,

[0252] (m) the heavy chain variable region CDR1 set forth in SEQ ID NO:13; the heavy chain variable region CDR2 set forth in SEQ ID NO:27; the heavy chain variable region CDR3 set forth in SEQ ID NO:41; the light chain variable region CDR1 set forth in SEQ ID NO:55; the light chain variable region CDR2 set forth in SEQ ID NO:69; and the light chain variable region CDR3 set forth in SEQ ID NO:83, or

[0253] (n) the heavy chain variable region CDR1 shown in SEQ ID NO:14; the heavy chain variable region CDR2 shown in SEQ ID NO:28; the heavy chain variable region CDR3 shown in SEQ ID NO:42; the light chain variable region CDR1 shown in SEQ ID NO:56; the light chain variable region CDR2 shown in SEQ ID NO:70; and the light chain variable region CDR3 shown in SEQ ID NO:84.

[0254] 24. An ActRII receptor antagonist for use according to any one of aspects 16-23, wherein the antibody comprises a full length heavy chain amino acid sequence having at least 95% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160.

[0255] 25. An ActRII receptor antagonist for use according to any one of aspects 16-24, wherein the antibody comprises a full length light chain amino acid sequence having at least 95% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: 155.

[0256] 26. An ActRII receptor antagonist for the method or use according to any one of aspects 16 to 25, wherein the antibody comprises:

[0257] (a) the variable heavy chain sequence shown in SEQ ID NO: 99 and the variable light chain sequence shown in SEQ ID NO: 85;

[0258] (b) the variable heavy chain sequence shown in SEQ ID NO: 100 and the variable light chain sequence shown in SEQ ID NO: 86;

[0259] (c) the variable heavy chain sequence shown in SEQ ID NO: 101 and the variable light chain sequence shown in SEQ ID NO: 87;

[0260] (d) the variable heavy chain sequence shown in SEQ ID NO: 102 and the variable light chain sequence shown in SEQ ID NO: 88;

[0261] (e) the variable heavy chain sequence shown in SEQ ID NO: 103 and the variable light chain sequence shown in SEQ ID NO: 89;

[0262] (f) the variable heavy chain sequence shown in SEQ ID NO: 104 and the variable light chain sequence shown in SEQ ID NO: 90;

[0263] (g) the variable heavy chain sequence shown in SEQ ID NO: 105 and the variable light chain sequence shown in SEQ ID NO: 91;

[0264] (h) the variable heavy chain sequence shown in SEQ ID NO: 106 and the variable light chain sequence shown in SEQ ID NO: 92;

[0265] (i) the variable heavy chain sequence set forth in SEQ ID NO: 107 and the variable light chain sequence set forth in SEQ ID NO: 93;

[0266] (j) the variable heavy chain sequence shown in SEQ ID NO: 108 and the variable light chain sequence shown in SEQ ID NO: 94;

[0267] (k) the variable heavy chain sequence shown in SEQ ID NO: 109 and the variable light chain sequence shown in SEQ ID NO: 95;

[0268] (1) the variable heavy chain sequence shown in SEQ ID NO: 110 and the variable light chain sequence shown in SEQ ID NO: 96;

[0269] (m) the variable heavy chain sequence shown in SEQ ID NO: 111 and the variable light chain sequence shown in SEQ ID NO: 97; or

[0270] (n) the variable heavy chain sequence shown in SEQ ID NO: 112 and the variable light chain sequence shown in SEQ ID NO: 98.

[0271] 27. An ActRII receptor antagonist for the method or use according to any one of aspects 16 to 26, wherein the antibody comprises:

[0272] (a) the heavy chain sequence shown in SEQ ID NO: 146 and the light chain sequence shown in SEQ ID NO: 141;

[0273] (b) the heavy chain sequence shown in SEQ ID NO: 147 and the light chain sequence shown in SEQ ID NO: 142;

[0274] (c) the heavy chain sequence shown in SEQ ID NO: 148 and the light chain sequence shown in SEQ ID NO: 143;

[0275] (d) the heavy chain sequence shown in SEQ ID NO: 149 and the light chain sequence shown in SEQ ID NO: 144;

[0276] (e) the heavy chain sequence shown in SEQ ID NO: 150 and the light chain sequence shown in SEQ ID NO: 145;

[0277] (f) the heavy chain sequence shown in SEQ ID NO: 156 and the light chain sequence shown in SEQ ID NO: 151;

[0278] (g) the heavy chain sequence shown in SEQ ID NO: 157 and the light chain sequence shown in SEQ ID NO: 152;

[0279] (h) the heavy chain sequence shown in SEQ ID NO: 158 and the light chain sequence shown in SEQ ID NO: 153;

[0280] (i) the heavy chain sequence shown in SEQ ID NO: 159 and the light chain sequence shown in SEQ ID NO: 154; or

[0281] (j) The heavy chain sequence shown in SEQ ID NO: 160 and the light chain sequence shown in SEQ ID NO: 155.

[0282] 28. An ActRII receptor antagonist for use according to any one of aspects 16 to 27, wherein the antibody comprised in the composition cross-blocks binding of at least one antibody according to aspect 27 to ActRIIB or is cross-blocked by binding of at least one antibody according to aspect 27 to ActRIIB.

[0283] 29. An ActRII receptor antagonist for use according to any one of aspects 16 to 28, wherein the antibody comprised in the composition has altered effector function by mutation of the Fc region.

[0284] 30. An ActRII receptor antagonist for use according to any one of aspects 16 to 29, wherein the antibody comprised in the composition binds to the epitope recognized by the antibody listed in aspects 26 to 27.

[0285] 31. An ActRII receptor antagonist for use according to any one of aspects 16 to 30, wherein the antibody is encoded by pBW522 (DSM22873) or pBW524 (DSM22874).

[0286] 32. Bimagrumab for use in the treatment and / or prevention of heart failure or for the treatment of structural and / or functional cardiac abnormalities associated with a condition selected from the group consisting of valvular heart disease, hypertension, coronary artery disease, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic and infectious agents, and other forms of hereditary or idiopathic cardiomyopathy, wherein bimagrumab is administered intravenously once every 4 weeks at a dose of about 3-10 mg / kg body weight.

[0287] 33. A composition comprising 150 mg / ml of bimagrumab for use in the treatment and / or prevention of heart failure or for the treatment of structural and / or functional cardiac abnormalities associated with a condition selected from the group consisting of valvular heart disease, hypertension, coronary artery disease, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress cardiomyopathy, toxic and infectious agents, and other forms of inherited or idiopathic cardiomyopathy.

[0288] 34. A single dosage form comprising 150 mg / ml of bimagrumab.

[0289] In another embodiment, a single dosage form, i.e., a vial, contains 100-200 mg / ml of bimagrumab, preferably 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200 mg / ml of bimagrumab.

[0290] 35. An infusion bag comprising an appropriate amount of bimagrumab from one or more vials diluted with a solution.

[0291] The solution is preferably a glucose solution.

[0292] In some additional embodiments, the ActRII receptor antagonist or anti-ActRII antibody (such as bimagrumab) is administered at a dose of about 1, 2, 3, 4, 5, 5, 6, 7, 8, 9, 10 mg / kg body weight.

[0293] Disclosed herein are ActRII receptor antagonists for use in the preparation of medicaments for treating and / or preventing heart failure and for treating structural and / or functional cardiac abnormalities associated with conditions such as valvular heart disease, hypertension, coronary artery disease, diabetes, aging, arrhythmias, peripartum cardiomyopathy, stress-induced cardiomyopathy, and other forms of inherited or idiopathic dilated cardiomyopathy.

[0294] In further embodiments, all aspects disclosed herein can be used in combination with any other aspects.

[0295] Various aspects of the present disclosure are described in more detail in the following subsections. Standard assays for evaluating the binding ability of antibodies to ActRII of various species are known in the art and include, for example, ELISA, Western blot, and RIA. The binding affinity of the antibodies can also be assessed by standard assays known in the art, such as by Biacore analysis or solution equilibrium titration. Surface plasmon resonance-based techniques (such as Biacore) can determine binding kinetics, which allows calculation of binding affinity.

[0296] Thus, an antibody that "inhibits" one or more of these ActRII functional properties (e.g., biochemical, immunochemical, cellular, physiological, or other biological activities, etc.) as determined according to methods known in the art and described herein should be understood to involve a statistically significant reduction in the specific activity relative to that observed in the absence of the antibody (e.g., or in the presence of a control antibody of irrelevant specificity). Antibodies that inhibit ActRII activity achieve such a statistically significant reduction of at least 10%, at least 50%, 80%, or 90% of the measured parameter, and in certain embodiments, the antibodies of the present disclosure can inhibit ActRIIB functional activity by greater than 95%, 98%, or 99%.

[0297] The ability or extent to which an antibody or other binding agent is able to interfere with the binding of another antibody or binding molecule to ActRII, and therefore whether it can be said to cross-block according to the present disclosure, can be determined using standard competitive binding assays. One suitable assay involves the use of Biacore technology (e.g., by using a BIAcore instrument (Biacore, Uppsala, Sweden)), which can use surface plasmon resonance technology to measure the extent of interaction. Another assay for measuring cross-blocking uses an ELISA-based method. Another assay uses FACS analysis, in which various antibodies are tested for competition for binding to ActRIIB-expressing cells.

[0298] According to the present disclosure, the cross-blocking antibodies or other binding agents according to the present disclosure bind to ActRII in the described BIAcore cross-blocking assay such that the recorded binding of the combination (cocktail) of the antibodies or binding agents is between 80% and 0.1% (e.g., 80% to 4%) of the maximal theoretical binding of the combination of the two antibodies or binding agents, specifically between 75% and 0.1% (e.g., 75% to 4%) of the maximal theoretical binding of the combination of the two antibodies or binding agents, more specifically between 70% and 0.1% (e.g., 70% to 4%) of the maximal theoretical binding of the combination of the two antibodies or binding agents, more specifically between 65% and 0.1% (e.g., 65% to 4%) of the maximal theoretical binding (as defined above) of the combination of the two antibodies or binding agents.

[0299] If the test antibody is able to cause a reduction in binding of the anti-ActRIIB antibody to ActRIIB by between 60% and 100%, specifically between 70% and 100%, more specifically between 80% and 100%, when compared to a positive control well (i.e., the same anti-ActRIIB antibody and ActRIIB, but without the "test" cross-blocking antibody), then the antibody is defined as cross-blocking the anti-ActRIIB antibodies of the present disclosure in an ELISA assay. Examples of cross-blocking antibodies cited herein are MOR08159 and MOR08213 (disclosed in WO2010 / 125003). Accordingly, the present disclosure provides compositions comprising antibodies that cross-block the binding of MOR08159 or MOR08213 to ActRIIB.

[0300] recombinant antibodies

[0301] Antibodies included in the compositions used in the present disclosure, e.g., ActRII antagonist antibodies such as bimagrumab, include isolated and structurally characterized human recombinant antibodies as described herein. H The amino acid sequences are shown in SEQ ID NO: 99 to SEQ ID NO: 112. The V LThe amino acid sequences are shown in SEQ ID NO: 85 to SEQ ID NO: 98, respectively. Examples of preferred full-length heavy chain amino acid sequences of the antibodies included in the compositions of the present invention are shown in SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160. Examples of preferred full-length light chain amino acid sequences of the antibodies included in the compositions of the present invention are shown in SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: 155, respectively. Other antibodies included in the compositions of the present invention include amino acids that have been mutated by amino acid deletion, insertion, or substitution, but have at least 60%, 70%, 80%, 90%, 95%, 97%, or 99% identity in the CDR regions to the CDR regions described in the above sequences. In some embodiments, it includes a mutant amino acid sequence in which no more than 1, 2, 3, 4 or 5 amino acids in the CDR region have been mutated by amino acid deletion, insertion or substitution when compared to the CDR region described by the above sequence.

[0302] In addition, variable heavy chain parent nucleotide sequences are shown in SEQ ID NO: 127 to SEQ ID NO: 140. Variable light chain parent nucleotide sequences are shown in SEQ ID NO: 113 to SEQ ID NO: 126. Full-length light chain nucleotide sequences optimized for expression in mammalian cells are shown in SEQ ID NO: 161 to SEQ ID NO: 165 and SEQ ID NO: 171 to SEQ ID NO: 175. Full-length heavy chain nucleotide sequences optimized for expression in mammalian cells are shown in SEQ ID NO: 166 to SEQ ID NO: 170 and SEQ ID NO: 176 to SEQ ID NO: 180. Other antibodies included in the compositions of the present invention include mutated amino acids or are encoded by mutated nucleic acids, but the mutated amino acids or mutated nucleic acids have a percent identity of at least 60 or more (i.e., 80, 90, 95, 97, 99 or more) with the above sequences. In some embodiments, the antibody comprises a mutant amino acid sequence wherein no more than 1, 2, 3, 4, or 5 amino acids in the variable region have been mutated by amino acid deletion, insertion, or substitution when compared to the variable region depicted in the above sequence.

[0303] Since each of these antibodies binds to the same epitope and is descended from the same parent antibody, V H 、V L, full-length light chain, and full-length heavy chain sequences (nucleotide and amino acid sequences) can be "mixed and matched" to generate other anti-ActRIIB binding molecules of the present disclosure. Such "mixed and matched" antibodies can be tested for ActRIIB binding using the binding assays described above and well-known methods (e.g., ELISA). When these chains are mixed and matched, the binding activity of the antibodies from a particular V H / V L Paired V H The sequence should be replaced by a structurally similar V H Likewise, the full-length heavy chain sequence from a particular full-length heavy chain / full-length light chain pairing should be replaced with a structurally similar full-length heavy chain sequence. H / V L Paired V L The sequence should be replaced by a structurally similar V L Sequence replacement. Similarly, the full-length chain sequence from a particular full-length heavy chain / full-length light chain pairing should be replaced with a structurally similar full-length chain sequence. Thus, in one aspect, the present disclosure provides a composition comprising a recombinant anti-ActRII antibody or antigen-binding region thereof having: a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 99 to SEQ ID NO: 112; and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 85 to SEQ ID NO: 98.

[0304] In another aspect, the present disclosure provides a composition comprising:

[0305] (i) an isolated recombinant anti-ActRII antibody having: a full-length heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 99 to SEQ ID NO: 112; and a full-length light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 85 to SEQ ID NO: 98, or

[0306] (ii) a functional protein comprising the antigen-binding portion of the isolated recombinant anti-ActRII antibody.

[0307] In another aspect, the present disclosure provides a composition comprising:

[0308] (i) an isolated recombinant anti-ActRII antibody having: a full-length heavy chain encoded by a nucleotide sequence optimized for expression in mammalian cells, the nucleotide sequence selected from the group consisting of SEQ ID NO: 127 to SEQ ID NO: 140; and a full-length light chain encoded by a nucleotide sequence optimized for expression in mammalian cells, the nucleotide sequence selected from the group consisting of SEQ ID NO: 113 to SEQ ID NO: 126, or

[0309] (ii) a functional protein comprising the antigen-binding portion of the isolated recombinant anti-ActRII antibody.

[0310] The V of the antibody contained in the composition of the present invention H Examples of the amino acid sequences of CDR1 are shown in SEQ ID NO: 1 to SEQ ID NO: 14. H The amino acid sequences of CDR2 are shown in SEQ ID NO: 15 to SEQ ID NO: 28. H The amino acid sequences of CDR3 are shown in SEQ ID NO: 29 to SEQ ID NO: 42. L The amino acid sequences of CDR1 are shown in SEQ ID NO: 43 to SEQ ID NO: 56. L The amino acid sequences of CDR2 are shown in SEQ ID NO: 57 to SEQ ID NO: 70. LThe amino acid sequences of CDR3 are shown in SEQ ID NO: 71 to SEQ ID NO: 84. The Kabat system is used to delineate CDR regions (Kabat, EA et al., 1991 Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). An alternative method for determining CDR regions uses a method designed by Chothia (Chothia et al., 25 1989, Nature, 342: 877-883). The Chothia definition is based on the position of the structural loop regions. However, due to variations in the numbering system used by Chothia (see, for example, http: / / www.biochem.ucl.ac.uk / ~martin / abs / Generallnfo.html and http: / / www.bioinf.org.uk / abs / ), this system is now less commonly used. Other systems exist for defining CDRs, and these systems are also mentioned on these two websites.

[0311] Given that each of these antibodies can bind to ActRII and that the antigen-binding specificity is primarily provided by the CDR1, CDR2, and CDR3 regions, V H CDR1, V H CDR2 and V H CDR3 sequence and V L CDR1, V L CDR2 and V L CDR3 sequences can be "mixed and matched" (i.e., CDRs from different antibodies can be mixed and matched, with each antibody containing V H CDR1, V H CDR2 and V H CDR3 and V L CDR1, V L CDR2 and V L CDR3, thereby generating other anti-ActRII binding molecules of the present disclosure. Such "mixed and matched" antibodies can be tested for ActRIIB binding using the binding assays described above and in the Examples (e.g., ELISA). H When CDR sequences are mixed and matched, H The CDR1, CDR2 and / or CDR3 sequences of the sequence should be replaced by CDR sequences with similar structures. L When CDR sequences are mixed and matched,L The CDR1, CDR2 and / or CDR3 sequences of the monoclonal antibodies shown in the present invention should be replaced by structurally similar CDR sequences. It is obvious to those of ordinary skill that by replacing one or more CDR sequences with structurally similar sequences from the CDR sequences of the monoclonal antibodies shown in the present invention, the CDR1, CDR2 and / or CDR3 sequences of the monoclonal antibodies shown in the present invention can be replaced by structurally similar CDR sequences. H and / or V L CDR region sequences can generate new V H Sequence and V L sequence.

[0312] The anti-ActRII antibodies or antigen-binding regions thereof included in the disclosed compositions have: a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14; a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28; a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: 42; a light chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 43 to SEQ ID NO: 56; a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 57 to SEQ ID NO: 70; and a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 71 to SEQ ID NO: 84.

[0313] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO: 1; the heavy chain variable region CDR2 shown in SEQ ID NO: 15; the heavy chain variable region CDR3 shown in SEQ ID NO: 29; the light chain variable region CDR1 shown in SEQ ID NO: 43; the light chain variable region CDR2 shown in SEQ ID NO: 57; and the light chain variable region CDR3 shown in SEQ ID NO: 71.

[0314] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:2; the heavy chain variable region CDR2 shown in SEQ ID NO:16; the heavy chain variable region CDR3 shown in SEQ ID NO:30; the light chain variable region CDR1 shown in SEQ ID NO:44; the light chain variable region CDR2 shown in SEQ ID NO:58; and the light chain variable region CDR3 shown in SEQ ID NO:72.

[0315] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO: 3; the heavy chain variable region CDR2 shown in SEQ ID NO: 17; the heavy chain variable region CDR3 shown in SEQ ID NO: 31; the light chain variable region CDR1 shown in SEQ ID NO: 45; the light chain variable region CDR2 shown in SEQ ID NO: 59; and the light chain variable region CDR3 shown in SEQ ID NO: 73.

[0316] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:4; the heavy chain variable region CDR2 shown in SEQ ID NO:18; the heavy chain variable region CDR3 shown in SEQ ID NO:32; the light chain variable region CDR1 shown in SEQ ID NO:46; the light chain variable region CDR2 shown in SEQ ID NO:60; and the light chain variable region CDR3 shown in SEQ ID NO:74.

[0317] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO: 5; the heavy chain variable region CDR2 shown in SEQ ID NO: 19; the heavy chain variable region CDR3 shown in SEQ ID NO: 33; the light chain variable region CDR1 shown in SEQ ID NO: 47; the light chain variable region CDR2 shown in SEQ ID NO: 61; and the light chain variable region CDR3 shown in SEQ ID NO: 75.

[0318] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO: 6; the heavy chain variable region CDR2 shown in SEQ ID NO: 20; the heavy chain variable region CDR3 shown in SEQ ID NO: 34; the light chain variable region CDR1 shown in SEQ ID NO: 48; the light chain variable region CDR2 shown in SEQ ID NO: 62; and the light chain variable region CDR3 shown in SEQ ID NO: 76.

[0319] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:7; the heavy chain variable region CDR2 shown in SEQ ID NO:21; the heavy chain variable region CDR3 shown in SEQ ID NO:35; the light chain variable region CDR1 shown in SEQ ID NO:49; the light chain variable region CDR2 shown in SEQ ID NO:63; and the light chain variable region CDR3 shown in SEQ ID NO:77.

[0320] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO: 8; the heavy chain variable region CDR2 shown in SEQ ID NO: 22; the heavy chain variable region CDR3 shown in SEQ ID NO: 36; the light chain variable region CDR1 shown in SEQ ID NO: 50; the light chain variable region CDR2 shown in SEQ ID NO: 64; and the light chain variable region CDR3 shown in SEQ ID NO: 78.

[0321] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:9; the heavy chain variable region CDR2 shown in SEQ ID NO:23; the heavy chain variable region CDR3 shown in SEQ ID NO:37; the light chain variable region CDR1 shown in SEQ ID NO:51; the light chain variable region CDR2 shown in SEQ ID NO:65; and the light chain variable region CDR3 shown in SEQ ID NO:79.

[0322] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:10; the heavy chain variable region CDR2 shown in SEQ ID NO:24; the heavy chain variable region CDR3 shown in SEQ ID NO:38; the light chain variable region CDR1 shown in SEQ ID NO:52; the light chain variable region CDR2 shown in SEQ ID NO:66; and the light chain variable region CDR3 shown in SEQ ID NO:80.

[0323] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:11; the heavy chain variable region CDR2 shown in SEQ ID NO:25; the heavy chain variable region CDR3 shown in SEQ ID NO:39; the light chain variable region CDR1 shown in SEQ ID NO:53; the light chain variable region CDR2 shown in SEQ ID NO:67; and the light chain variable region CDR3 shown in SEQ ID NO:81.

[0324] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:12; the heavy chain variable region CDR2 shown in SEQ ID NO:26; the heavy chain variable region CDR3 shown in SEQ ID NO:40; the light chain variable region CDR1 shown in SEQ ID NO:54; the light chain variable region CDR2 shown in SEQ ID NO:68; and the light chain variable region CDR3 shown in SEQ ID NO:82.

[0325] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:13; the heavy chain variable region CDR2 shown in SEQ ID NO:27; the heavy chain variable region CDR3 shown in SEQ ID NO:41; the light chain variable region CDR1 shown in SEQ ID NO:55; the light chain variable region CDR2 shown in SEQ ID NO:69; and the light chain variable region CDR3 shown in SEQ ID NO:83.

[0326] In one embodiment, the antibody contained in the composition of the present invention comprises: the heavy chain variable region CDR1 shown in SEQ ID NO:14; the heavy chain variable region CDR2 shown in SEQ ID NO:28; the heavy chain variable region CDR3 shown in SEQ ID NO:42; the light chain variable region CDR1 shown in SEQ ID NO:56; the light chain variable region CDR2 shown in SEQ ID NO:70; and the light chain variable region CDR3 shown in SEQ ID NO:84.

[0327] In one embodiment, the present disclosure provides a composition comprising an antibody comprising: (a) a variable heavy chain sequence set forth in SEQ ID NO: 85 and a variable light chain sequence set forth in SEQ ID NO: 99; (b) a variable heavy chain sequence set forth in SEQ ID NO: 86 and a variable light chain sequence set forth in SEQ ID NO: 100; (c) a variable heavy chain sequence set forth in SEQ ID NO: 87 and a variable light chain sequence set forth in SEQ ID NO: 101; (d) a variable heavy chain sequence set forth in SEQ ID NO: 88 and a variable light chain sequence set forth in SEQ ID NO: 102; (e) a variable heavy chain sequence set forth in SEQ ID NO: 89 and a variable light chain sequence set forth in SEQ ID NO: 103; (f) a variable heavy chain sequence set forth in SEQ ID NO: 90 and a variable light chain sequence set forth in SEQ ID NO: 104; (g) a variable heavy chain sequence set forth in SEQ ID NO: 91 and a variable light chain sequence set forth in SEQ ID NO: 105; (h) a variable heavy chain sequence set forth in SEQ ID NO: 92 and a variable light chain sequence set forth in SEQ ID NO: 106; (i) a variable heavy chain sequence set forth in SEQ ID NO: 88 and a variable light chain sequence set forth in SEQ ID NO: 102. (i) the variable heavy chain sequence shown in SEQ ID NO:93 and the variable light chain sequence shown in SEQ ID NO:107; (j) the variable heavy chain sequence shown in SEQ ID NO:94 and the variable light chain sequence shown in SEQ ID NO:108; (k) the variable heavy chain sequence shown in SEQ ID NO:95 and the variable light chain sequence shown in SEQ ID NO:109; (l) the variable heavy chain sequence shown in SEQ ID NO:96 and the variable light chain sequence shown in SEQ ID NO:110; (m) the variable heavy chain sequence shown in SEQ ID NO:97 and the variable light chain sequence shown in SEQ ID NO:111; or (n) the variable heavy chain sequence shown in SEQ ID NO:98 and the variable light chain sequence shown in SEQ ID NO:112.

[0328] In one embodiment, the present disclosure provides a composition comprising an antibody comprising: (a) a heavy chain sequence set forth in SEQ ID NO: 146 and a light chain sequence set forth in SEQ ID NO: 141; (b) a heavy chain sequence set forth in SEQ ID NO: 147 and a light chain sequence set forth in SEQ ID NO: 142; (c) a heavy chain sequence set forth in SEQ ID NO: 148 and a light chain sequence set forth in SEQ ID NO: 143; (d) a heavy chain sequence set forth in SEQ ID NO: 149 and a light chain sequence set forth in SEQ ID NO: 144; (e) a heavy chain sequence set forth in SEQ ID NO: 150 and a light chain sequence set forth in SEQ ID NO: 145; (f) a heavy chain sequence set forth in SEQ ID NO: 156 and a light chain sequence set forth in SEQ ID NO: 151; (g) a heavy chain sequence set forth in SEQ ID NO: 157 and a light chain sequence set forth in SEQ ID NO: 152; (h) a heavy chain sequence set forth in SEQ ID NO: 158 and a light chain sequence set forth in SEQ ID NO: 153; (i) The heavy chain sequence shown in SEQ ID NO: 159 and the light chain sequence shown in SEQ ID NO: 154; or (j) the heavy chain sequence shown in SEQ ID NO: 160 and the light chain sequence shown in SEQ ID NO: 155.

[0329] As used herein, a human antibody comprises a heavy chain variable region or a light chain variable region or a full-length heavy chain or a full-length light chain. If the variable region or full-length chain of the antibody is obtained from a system that uses human germline immunoglobulin genes, then the heavy chain variable region or light chain variable region or full-length heavy chain or full-length light chain is the "product" or "derived from" a specific germline sequence. Such systems include immunizing transgenic mice carrying human immunoglobulin genes with an antigen of interest, or screening a human immunoglobulin gene library displayed on phage with an antigen of interest. Thus, by comparing the amino acid sequence of a human antibody with the amino acid sequence of a human germline immunoglobulin and selecting the human germline immunoglobulin sequence that is closest in sequence to the sequence of the human antibody (i.e., the greatest % identity), it is possible to identify a human antibody as a "product" or "derived from" a human germline immunoglobulin sequence. Human antibodies that are the "product" or "derived from" a particular human germline immunoglobulin sequence may contain amino acid differences compared to the germline sequence due to, for example, naturally occurring somatic mutations or intentionally introduced site-directed mutations. However, the amino acid sequence of the selected human antibody is typically at least 90% identical to the amino acid sequence encoded by the human germline immunoglobulin gene and contains amino acid residues that identify the human antibody as human when compared to the germline immunoglobulin amino acid sequences of other species (e.g., mouse germline sequences). In some cases, the amino acid sequence of the human antibody may be at least 80%, 90%, or at least 95%, or even at least 96%, 97%, 98%, or 99% identical to the amino acid sequence encoded by the germline immunoglobulin gene. Typically, a human antibody derived from a particular human germline sequence will exhibit no more than 10 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene. In some cases, a human antibody may exhibit no more than 5, or even no more than 4, 3, 2, or 1 amino acid differences from the amino acid sequence encoded by the germline immunoglobulin gene.

[0330] In one embodiment, the antibody comprised in the compositions of the present disclosure is the antibody encoded by pBW522 or pBW524 (deposited on August 18, 2009 at DSMZ, Inhoffenstr. 7B, D-38124 Braunschweig, Germany under the deposit numbers DSM 22873 and DSM 22874, respectively).

[0331] Homologous antibodies

[0332] In another embodiment, the antibodies included in the compositions of the invention have a full-length heavy chain amino acid sequence and a full-length light chain amino acid sequence; a full-length heavy chain nucleotide sequence and a full-length light chain nucleotide sequence, a variable region heavy chain amino acid sequence and a variable region light chain amino acid sequence, or a variable region heavy chain nucleotide sequence and a variable region light chain nucleotide sequence that are homologous to the amino acid sequence and nucleotide sequence of the antibodies described herein, and wherein the antibodies retain the desired functional properties of the anti-ActRIIB antibodies of the present disclosure.

[0333] For example, the present disclosure provides a composition comprising an isolated recombinant anti-ActRIIB antibody (or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRIIB antibody), wherein the isolated recombinant anti-ActRIIB antibody (or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRIIB antibody) comprises a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises an amino acid sequence that is at least 80%, or at least 90% (preferably at least 95%, 97% or 99%) identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 99 to SEQ ID NO: 112; the light chain variable region comprises an amino acid sequence that is at least 80%, or at least 90% (preferably at least 95%, 97% or 99%) identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 85 to SEQ ID NO: NO:98, or an amino acid sequence that is at least 80%, or at least 90% (preferably at least 95%, 97% or 99%) identical to the amino acid sequence of the group consisting of SEQ ID NO:98; or, the composition comprises a recombinant anti-ActRIIB antibody (or a functional protein comprising the antigen-binding portion of a recombinant anti-ActRIIB antibody), wherein the recombinant anti-ActRIIB antibody (or a functional protein comprising the antigen-binding portion of a recombinant anti-ActRIIB antibody) comprises a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises no more than 5 amino acids, or no more than 4 amino acids, or no more than 3 amino acids, or no more than 2 amino acids, or no more than 1 amino acid change compared to the amino acid sequence selected from the group consisting of SEQ ID NO:99 to SEQ ID NO:112; the light chain variable region comprises no more than 5 amino acids, or no more than 4 amino acids, or no more than 3 amino acids, or no more than 2 amino acids, or no more than 1 amino acid change compared to the amino acid sequence selected from the group consisting of SEQ ID NO:85 to SEQ ID NO: The amino acid sequence of the group consisting of ID NO: 98 has no more than 5 amino acids, or no more than 4 amino acids, or no more than 3 amino acids, or no more than 2 amino acids, or no more than 1 amino acid change compared to the amino acid sequence of the group consisting of ID NO: 98, and the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo, (ii) reduces inhibition of muscle differentiation through a Smad-dependent pathway, and / or (iii) does not induce hematological changes, in particular, no changes in RBC. As used herein, the term "alteration / change" refers to insertion, deletion and / or substitution.

[0334] In another example, the present disclosure provides a composition comprising an isolated recombinant anti-ActRII antibody (or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRII antibody), wherein the isolated recombinant anti-ActRII antibody (or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRII antibody) comprises a full-length heavy chain and a full-length light chain, wherein: the full-length heavy chain comprises an amino acid sequence that is at least 80%, or at least 90% (preferably at least 95%, 97%, or 99%) identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160; and the full-length light chain comprises an amino acid sequence that is at least 80%, or at least 90% (preferably at least 95%, 97%, or 99%) identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: NO: 156 to SEQ ID NO: 160; and The amino acid sequence of the group consisting of NO: 155 has no more than 5 amino acids, or no more than 4 amino acids, or no more than 3 amino acids, or no more than 2 amino acids, or no more than 1 amino acid change compared to the amino acid sequence of NO: 155, and the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo, (ii) reduces inhibition of muscle differentiation through a Smad-dependent pathway, and / or (iii) does not induce hematological changes, in particular, no changes in RBC. Preferably, such antibodies bind to the ligand binding domain of ActRIIB and / or ActRIIA. As used herein, the term "alteration / change" refers to insertion, deletion, and / or substitution.

[0335] In another example, the present disclosure provides a composition comprising an isolated recombinant anti-ActRII antibody (or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRII antibody), wherein the isolated recombinant anti-ActRII antibody (or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRII antibody) comprises a full-length heavy chain and a full-length light chain, wherein: the full-length heavy chain is encoded by a nucleotide sequence that is at least 80%, or at least 90% (preferably at least 95%, 97% or 99%) identical to a nucleotide sequence selected from the group consisting of SEQ ID NO: 166 to SEQ ID NO: 170 and SEQ ID NO: 176 to SEQ ID NO: 180; and the full-length light chain is encoded by a nucleotide sequence that is at least 80%, or at least 90% (preferably at least 95%, 97% or 99%) identical to a nucleotide sequence selected from the group consisting of SEQ ID NO: 161 to SEQ ID NO: 165 and SEQ ID NO: 171 to SEQ ID NO: NO: 175; or, the composition comprises a recombinant anti-ActRIIB antibody (or a functional protein comprising an antigen-binding portion of a recombinant anti-ActRIIB antibody), wherein the recombinant anti-ActRIIB antibody (or a functional protein comprising an antigen-binding portion of a recombinant anti-ActRIIB antibody) comprises a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises no more than 5 amino acids, or no more than 4 amino acids, or no more than 3 amino acids, or no more than 2 amino acids, or no more than 1 amino acid change compared to the amino acid sequence selected from the group consisting of SEQ ID NO: 166 to SEQ ID NO: 170 and SEQ ID NO: 176 to SEQ ID NO: 180; the light chain variable region comprises no more than 1 amino acid change compared to the amino acid sequence selected from the group consisting of SEQ ID NO: 161 to SEQ ID NO: 165 and SEQ ID NO: 171 to SEQ ID NO: The amino acid sequence of the group consisting of NO: 175 has no more than 5 amino acids, or no more than 4 amino acids, or no more than 3 amino acids, or no more than 2 amino acids, or no more than 1 amino acid change compared to the amino acid sequence of NO: 175, and the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo, (ii) reduces inhibition of muscle differentiation through a Smad-dependent pathway, and / or (iii) does not induce hematological changes, in particular, there is no change in RBC. Preferably, such antibodies bind to the ligand binding domain of ActRIIB. As used herein, the term "alteration / change" refers to insertion, deletion, and / or substitution.

[0336] In various embodiments, the antibodies included in the compositions of the present invention may exhibit one or more, two or more, or three of the functional properties discussed above. The antibodies may be, for example, human antibodies, humanized antibodies, or chimeric antibodies. Preferably, the antibodies are fully human IgG1 antibodies.

[0337] In other embodiments, V H and / or V L The amino acid sequence may be at least 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the above sequences. In other embodiments, except for the amino acid substitution at no more than 1, 2, 3, 4 or 5 amino acid positions, V H and / or V L The amino acid sequences may be identical. H and V L The V region has high (ie, 80% or greater) identity H and V L Antibodies to the regions can be obtained by mutagenizing the nucleic acid molecules SEQ ID NO: 127 to SEQ ID NO: 140 and SEQ ID NO: 113 to SEQ ID NO: 126, respectively (e.g., site-directed mutagenesis or PCR-mediated mutagenesis), and then using the functional assays described herein to test whether the encoded altered antibodies retain function (i.e., the above-mentioned functions).

[0338] In other embodiments, the full-length heavy chain and / or full-length light chain amino acid sequence can be at least 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the above-mentioned sequences, or can be identical to the above-mentioned sequences except for amino acid changes in no more than 1, 2, 3, 4 or 5 amino acid positions. Antibodies having full-length heavy chains and full-length light chains that are highly identical (i.e., at least 80% or more) to the full-length heavy chains set forth in any one of SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160, and the full-length light chains set forth in any one of SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: 155, respectively, can be obtained by mutagenizing (e.g., site-directed mutagenesis or PCR-mediated mutagenesis) the nucleic acid molecules SEQ ID NO: 166 to SEQ ID NO: 170 and SEQ ID NO: 176 to SEQ ID NO: 180, and SEQ ID NO: 161 to SEQ ID NO: 165 and SEQ ID NO: 171 to SEQ ID NO: 175, respectively, and then testing whether the encoded altered antibodies retain function (i.e., the functions described above) using the functional assays described herein.

[0339] In other embodiments, the full-length heavy chain and / or full-length light chain nucleotide sequences may be at least 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the above sequences.

[0340] In other embodiments, the variable region of the heavy and / or light chain nucleotide sequence may be at least 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the above sequences, or may be identical to the above sequences except for amino acid changes in no more than 1, 2, 3, 4 or 5 amino acid positions.

[0341] As used herein, the percent identity between two sequences is a function of the number of identical positions that the sequences share (i.e., the number of % identity=identical positions / position totals × 100), taking into account the number of spaces that need to be introduced to realize the optimal comparison of the two sequences and the length of each space. As described below, a mathematical algorithm can be used to complete the comparison of the sequence and to the determination of the percent identity between the two sequences.

[0342] The percent identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4: 11-17, 1988), which has been incorporated into the ALIGN program (version 2.0), using a PAM 120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol, Biol. 48: 444-453, 1970) algorithm, which has been incorporated into the GAP program in the GCG software package (available at http: / / www.gcg.com), using a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.

[0343] Antibodies with conservative modifications

[0344] In certain embodiments, the antibodies included in the compositions of the present invention have a heavy chain variable region comprising a CDR1, CDR2, and CDR3 sequence and a light chain variable region comprising a CDR1, CDR2, and CDR3 sequence, wherein one or more of these CDR sequences has a specified amino acid sequence based on an antibody described herein, or a variant sequence thereof comprising 1, 2, 3, 4, or 5 amino acid changes, or conservative modifications thereof, and wherein the antibody retains the desired functional properties of the anti-ActRIIB antibodies of the present disclosure.

[0015] Accordingly, the present disclosure provides a composition comprising an isolated recombinant anti-ActRIIB antibody or a functional protein comprising an antigen-binding portion of an isolated recombinant anti-ActRIIB antibody, wherein the isolated recombinant anti-ActRIIB antibody or the functional protein comprising the antigen-binding portion of an isolated recombinant anti-ActRIIB antibody comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences, and a light chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein: the heavy chain variable region CDR1 amino acid sequence is selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14, or variant sequences thereof comprising 1, 2, 3, 4, or 5 amino acid changes, and conservative modifications thereof; the heavy chain variable region CDR2 amino acid sequence is selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28, or variant sequences thereof comprising 1, 2, 3, 4, or 5 amino acid changes, and conservative modifications thereof; the heavy chain variable region CDR3 amino acid sequence is selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: NO:42 or a variant sequence thereof comprising 1, 2, 3, 4 or 5 amino acid changes, and conservative modifications thereof; the light chain variable region CDR1 amino acid sequence is selected from SEQ ID NO:43 to SEQ ID NO:56 or a variant sequence thereof comprising 1, 2, 3, 4 or 5 amino acid changes, and conservative modifications thereof; the light chain variable region CDR2 amino acid sequence is selected from SEQ ID NO:57 to SEQ ID NO:70 or a variant sequence thereof comprising 1, 2, 3, 4 or 5 amino acid changes, and conservative modifications thereof; the light chain variable region CDR3 amino acid sequence is selected from SEQ ID NO:71 to SEQ ID NO:84 or a variant sequence thereof comprising 1, 2, 3, 4 or 5 amino acid changes, and conservative modifications thereof. Preferably, the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo, (ii) reduces inhibition of muscle differentiation through a Smad-dependent pathway, and / or (iii) does not induce hematological changes, in particular, no changes in RBCs.

[0345] In various embodiments, the antibody may exhibit one or both of the functional properties listed above. Such an antibody may be, for example, a human antibody, a humanized antibody, or a chimeric antibody.

[0346] In other embodiments, the antibodies included in the compositions of the present invention that are optimized for expression in mammalian cells have full-length heavy chain sequences and full-length light chain sequences, wherein one or more of these sequences have a specified amino acid sequence based on an antibody described herein or a conservative modification thereof, and wherein the antibody retains the desired functional properties of the anti-ActRIIB antibodies of the present disclosure. Accordingly, the present disclosure provides a composition comprising an isolated monoclonal anti-ActRII antibody that is optimized for expression in mammalian cells, the isolated monoclonal anti-ActRII antibody consisting of a full-length heavy chain and a full-length light chain, wherein: the full-length heavy chain has an amino acid sequence selected from the group consisting of SEQ ID NO: 146 to SEQ ID NO: 150 and SEQ ID NO: 156 to SEQ ID NO: 160, or variant sequences thereof comprising 1, 2, 3, 4, or 5 amino acid changes, and conservative modifications thereof; and the full-length light chain has an amino acid sequence selected from the group consisting of SEQ ID NO: 141 to SEQ ID NO: 145 and SEQ ID NO: 151 to SEQ ID NO: NO:155 or its variant sequence comprising 1, 2, 3, 4 or 5 amino acid changes and conservative modifications thereof; and the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo, (ii) reduces inhibition of muscle differentiation through a Smad-dependent pathway, and / or (iii) does not induce hematological changes, in particular no changes in RBCs.

[0347] In various embodiments, the antibody may exhibit one or both of the functional properties listed above. Such an antibody may be, for example, a human antibody, a humanized antibody, or a chimeric antibody.

[0348] As used herein, the term "conservative sequence modification" refers to amino acid modifications that do not significantly affect or alter the binding characteristics of the antibody containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into the antibodies of the present disclosure by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis.

[0349] Conservative amino acid substitutions are amino acid substitutions in which amino acid residues are replaced by amino acid residues with similar side chains. Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), amino acids with non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Therefore, one or more amino acid residues in the CDR regions of antibodies of the present disclosure can be replaced with other amino acid residues from the same side chain family, and functional assays described herein can be used to test whether the antibody of the change retains function.

[0350] Antibodies that bind to the same epitope as the anti-ActRII antibodies included in the disclosed compositions

[0351] In another embodiment, the present disclosure provides compositions comprising antibodies that bind to the same epitope as the various specific anti-ActRII antibodies described herein. All antibodies described in the Examples that block myostatin binding to ActRIIA and ActRIIB bind with high affinity to one of the epitopes in ActRIIA and ActRIIB that is contained between amino acids 19-134 of SEQ ID NO: 181.

[0352] Thus, additional antibodies can be identified based on their ability to cross-compete with other antibodies of the present disclosure (e.g., competitively inhibit the binding of other antibodies of the present disclosure in a statistically significant manner) in a standard ActRIIB binding assay. The ability of a test antibody to inhibit the binding of an antibody contained in the composition of the present invention to human ActRIIB indicates that the test antibody can compete with the antibody for binding to human ActRIIB; according to non-limiting theory, such an antibody may bind to the same or related (e.g., structurally similar or spatially adjacent) epitope on human ActRIIB as the antibody it competes with. In a certain embodiment, the antibody that binds to the same epitope on human ActRIIA and ActRIIA as the antibody contained in the composition of the present invention is a human recombinant antibody. Such human recombinant antibodies can be prepared and isolated as described in the Examples. Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO: 85 and a variable light chain sequence set forth in SEQ ID NO: 99 and / or compete for binding with an antibody having a variable heavy chain sequence set forth in SEQ ID NO: 85 and a variable light chain sequence set forth in SEQ ID NO: 99.

[0353] Thus, the present disclosure provides compositions comprising antibodies that bind to the epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:86 and a variable light chain sequence set forth in SEQ ID NO:100.

[0354] Thus, the present disclosure provides compositions comprising an antibody that binds to an epitope recognized by an antibody having a variable heavy chain sequence as set forth in SEQ ID NO: 87 and a variable light chain sequence as set forth in SEQ ID NO: 101. Thus, the present disclosure provides compositions comprising an antibody that binds to an epitope recognized by an antibody having a variable heavy chain sequence as set forth in SEQ ID NO: 88 and a variable light chain sequence as set forth in SEQ ID NO: 102. Thus, the present disclosure provides compositions comprising an antibody that binds to an epitope recognized by an antibody having a variable heavy chain sequence as set forth in SEQ ID NO: 89 and a variable light chain sequence as set forth in SEQ ID NO: 103.

[0355] Thus, the present disclosure provides compositions comprising antibodies that bind to the epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:90 and a variable light chain sequence set forth in SEQ ID NO:104.

[0356] Thus, the present disclosure provides compositions comprising antibodies that bind to the epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:91 and a variable light chain sequence set forth in SEQ ID NO:105.

[0357] Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:92 and a variable light chain sequence set forth in SEQ ID NO:106.

[0358] Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:93 and a variable light chain sequence set forth in SEQ ID NO:107.

[0359] Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:94 and a variable light chain sequence set forth in SEQ ID NO:108.

[0360] Thus, the present disclosure provides compositions comprising antibodies that bind to the epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:95 and a variable light chain sequence set forth in SEQ ID NO:109.

[0361] Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:96 and a variable light chain sequence set forth in SEQ ID NO:110.

[0362] Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:97 and a variable light chain sequence set forth in SEQ ID NO:111.

[0363] Thus, the present disclosure provides compositions comprising antibodies that bind to an epitope recognized by an antibody having a variable heavy chain sequence set forth in SEQ ID NO:98 and a variable light chain sequence set forth in SEQ ID NO:112.

[0364] Following more detailed epitope mapping experiments, the binding regions of the preferred antibodies of the compositions of the invention have been more clearly defined.

[0365] Thus, the present disclosure provides compositions comprising an antibody that binds to an epitope comprising amino acids 78-83 (WLDDFN - SEQ ID NO: 188) of SEQ ID NO: 181. The present disclosure also provides compositions comprising an antibody that binds to an epitope comprising amino acids 76-84 (GCWLDDFNC - SEQ ID NO: 186) of SEQ ID NO: 181.

[0366] The present disclosure also provides compositions comprising an antibody that binds to an epitope comprising amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY—SEQ ID NO: 190).

[0367] The present disclosure also provides compositions comprising an antibody that binds to an epitope comprising amino acids 52-56 (EQDKR - SEQ ID NO: 189) of SEQ ID NO: 181. The present disclosure also provides compositions comprising an antibody that binds to an epitope comprising amino acids 49-63 (CEGEQDKRLHCYASW - SEQ ID NO: 187) of SEQ ID NO: 181.

[0368] The present disclosure also provides compositions comprising an antibody that binds to an epitope comprising or consisting of amino acids 29-41 of SEQ ID NO: 181 (CIYYNANWELERT-SEQ ID NO: 191).

[0369] The present disclosure also provides compositions comprising an antibody that binds to an epitope comprising or consisting of amino acids 100-110 of SEQ ID NO: 181 (YFCCCEGNFCN-SEQ ID NO: 192).

[0370] The present disclosure also provides compositions comprising antibodies that bind to epitopes consisting of these sequences or comprising a combination of these epitope regions.

[0371] Thus, the present disclosure also provides compositions comprising an antibody that binds to an epitope comprising or consisting of amino acids 78-83 of SEQ ID NO: 181 (WLDDFN) and amino acids 52-56 of SEQ ID NO: 181 (EQDKR).

[0372] Engineered and modified antibodies

[0373] Antibodies included in the compositions of the present invention may also be used with V H and / or V L Antibodies containing one or more of the sequences are prepared as starting materials to engineer modified antibodies that may have altered properties compared to the starting antibody. Antibodies can be engineered by modifying one or two variable regions (i.e., V H and / or V L ), for example, one or more residues within one or more CDR regions and / or or one or more residues within one or more framework regions. Additionally or alternatively, the antibody can be engineered by modifying residues within one or more constant regions, for example, to alter the effector functions of the antibody.

[0374] One type of variable region engineering that can be performed is CDR grafting. Antibodies interact with target antigens primarily through amino acid residues located in the six heavy and light chain complementary determining regions (CDRs). Therefore, the amino acid sequences within the CDRs are more diverse between antibodies than the sequences outside the CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, recombinant antibodies that mimic the properties of a specific naturally occurring antibody can be expressed by constructing expression vectors that include the CDR sequences from a specific naturally occurring antibody grafted onto framework sequences from a different antibody with different properties (see, e.g., Riechmann, L. et al., 1998 Nature 332:323-327; Jones, P. et al., 1986 Nature 321:522-525; Queen, C. et al., 1989 Proc. Natl. Acad. Sci. USA 86:10029-10033; U.S. Pat. No. 5,225,539 to Winter, and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.).

[0375] Accordingly, another embodiment of the present disclosure is directed to a composition comprising a monoclonal anti-ActRII antibody, or a functional protein comprising an antigen-binding portion of a monoclonal anti-ActRII antibody, comprising a heavy chain variable region comprising: a CDR1 sequence having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14; a CDR2 sequence having an amino acid sequence selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28; and a CDR3 sequence having an amino acid sequence selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: 42; and a light chain variable region comprising: a CDR1 sequence having an amino acid sequence selected from the group consisting of SEQ ID NO: 43 to SEQ ID NO: 56; a CDR2 sequence having an amino acid sequence selected from the group consisting of SEQ ID NO: 57 to SEQ ID NO: 70; and a CDR3 sequence selected from the group consisting of SEQ ID NO: 71 to SEQ ID NO: 84. Therefore, these antibodies contain the V H CDR sequences and V L CDR sequences, but may contain different framework sequences from these antibodies.

[0376] Such framework sequences can be obtained from public references or public DNA databases that include germline antibody gene sequences. For example, the germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database (available on the Internet at www.mrc-cpe.cam.ac.uk / vbase), as well as in Kabat, EA et al., [as described above]; Tomlinson, IM et al., 1992 J. fol. Biol. 227: 776-798; and Cox, JPL et al., 1994 Eur. J Immunol. 24: 827-836. Examples of framework sequences used in the antibodies of the present disclosure are those that are structurally similar to the framework sequences used for the selected antibodies of the present disclosure (e.g., the consensus sequences and / or framework sequences used for the monoclonal antibodies of the present disclosure). V H CDR1, V H CDR2 and V H CDR3 sequence and V L CDR1, V L CDR2 and V LThe CDR3 sequence is grafted onto a framework region having the same sequence as that found in the germline immunoglobulin gene from which the framework sequence is derived, or the CDR sequence can be grafted onto a framework region containing one or more mutations compared to the germline sequence. For example, it has been found that in certain circumstances, mutating residues within the framework region is beneficial for maintaining or enhancing the antigen-binding ability of the antibody (see, e.g., U.S. Patents 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.).

[0377] Another type of variable region modification is to make V H and / or V L Amino acid residue mutations within the CDR1, CDR2 and / or CDR3 regions, thereby improving one or more binding properties (e.g., affinity) of an antibody of interest, are referred to as "affinity maturation." Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce one or more mutations, and the impact on antibody binding or other functional properties of interest can be assessed in an in vitro or in vivo assay as described herein and provided in the Examples. Conservative modifications (as described above) can be introduced. Mutations can be amino acid substitutions, additions, or deletions. In addition, typically no more than one, two, three, four, or five residues are changed within the CDR regions.

[0378] Thus, in another embodiment, the present disclosure provides an isolated anti-ActRII monoclonal antibody, or a functional protein comprising an antigen-binding portion of an isolated anti-ActRII monoclonal antibody, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region has: V H CDR1 region, the V H The CDR1 region consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 14, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NO: 1 to SEQ ID NO: 14; H CDR2 region, the V H The CDR2 region has an amino acid sequence selected from the group consisting of SEQ ID NO: 15 to SEQ ID NO: 28, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NO: 15 to SEQ ID NO: 28; H CDR3 region, the V HThe CDR3 region has an amino acid sequence selected from the group consisting of SEQ ID NO: 29 to SEQ ID NO: 42, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NO: 29 to SEQ ID NO: 42; L CDR1 region; the heavy chain variable region has: the V L The CDR1 region has an amino acid sequence selected from the group consisting of SEQ ID NO: 43 to SEQ ID NO: 56, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NO: 43 to SEQ ID NO: 56; L CDR2 region, the V L The CDR2 region has an amino acid sequence consisting of the group consisting of SEQ ID NO: 52 to SEQ ID NO: 70, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NO: 52 to SEQ ID NO: 70; and V L CDR3 region, the V L The CDR3 region has an amino acid sequence of the group consisting of SEQ ID NO: 71 to SEQ ID NO: 84, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NO: 71 to SEQ ID NO: 84.

[0379] Camelid antibodies

[0380] Antibody proteins obtained from camels and members of the dromedary family (Camelus bactrianus and Camelus dromaderius), including New World members such as llama species (Lama paccos, Lama glama, and Lamavicugna), have been characterized in terms of size, structural complexity, and antigenicity to human subjects. Certain IgG antibodies from this mammalian family found in nature lack light chains and are therefore structurally distinct from the typical four-chain quaternary structure of antibodies from other animals, having two heavy chains and two light chains (see WO 94 / 04678).

[0381] A region of camelid antibodies identified as V HHSmall single variable domains (SBDs) can be obtained by genetically engineering small proteins with high affinity for their targets, thereby generating low molecular weight antibody-derived proteins known as “camelid nanobodies” (see US 5,759,808; Stijlemans, B. et al., 2004 J Biol Chem 279:1256-1261; Dumoulin, M. et al., 2003 Nature 424:783-788; Pleschberger, M. et al., 2003 Bioconjugate Chem 14:440-448; Cortez-Retamozo, V. et al., 2002 Int J Cancer 89:456-62; and Lauwereys, M. et al., 1998 EMBO J 17:3512-3520). Engineered libraries of camelid antibodies and antibody fragments are commercially available, for example from Ablynx, Ghent, Belgium. As with other non-human antibodies, the amino acid sequence of camelid antibodies can be recombinantly altered to more closely resemble human sequences, i.e., Nanobodies can be “humanized.” Thus, the naturally low antigenicity of camelid antibodies to humans can be further reduced.

[0382] The molecular weight of camelid nanobodies is approximately one-tenth that of a human IgG molecule, and the physical diameter of the protein is only a few nanometers. One consequence of their small size is that camelid nanobodies have the ability to bind to antigenic sites that are functionally invisible to larger antibody proteins, meaning that camelid nanobodies can be used as reagents for detecting antigens that are hidden using classical immunological techniques, as well as as potential therapeutic agents. Consequently, another consequence of their small size is that camelid nanobodies can inhibit by binding to specific sites in grooves or narrow crevices of target proteins, thus potentially providing the ability to more closely approximate the function of classical low-molecular-weight drugs than classical antibodies.

[0383] Low molecular weight and compact size further result in camel family nano antibodies being extremely heat stable, stable to extreme pH and proteolytic digestion, and poor antigenicity. Another result is that camel family nano antibodies easily move from the circulatory system to tissues, even across the blood-brain barrier and can treat diseases affecting nervous tissue. Nano antibodies can further promote drug transport across the blood-brain barrier (see US2004 / 0161738). The combination of these features and low antigenicity to humans shows huge therapeutic potential. In addition, these molecules can be fully expressed in prokaryotic cells such as E. coli, and are expressed as fusion proteins and are functional using phage.

[0384] Thus, in one embodiment, the present disclosure relates to a composition comprising a camelid antibody or nanobody with high affinity for ActRIIB. In certain embodiments herein, the camelid antibody or nanobody is naturally produced in a camelid, i.e., produced by a camelid following immunization with ActRIIB or a peptide fragment thereof, using the techniques described herein for other antibodies. Alternatively, the anti-ActRIIB camelid nanobody is engineered, i.e., produced by selecting from a phage library displaying appropriately mutagenized camelid nanobody proteins using a panning procedure targeting ActRIIB, as described in the Examples herein. The engineered nanobody can be further customized by genetic engineering so that the half-life in the recipient subject is from 45 minutes to two weeks. In a specific embodiment, the camelid antibody or nanobody is obtained by transplanting the CDR sequences of the heavy or light chain of a human antibody of the present disclosure into a nanobody or single domain antibody framework sequence, as described, for example, in WO94 / 04678.

[0385] Non-antibody scaffolds

[0386] Known non-immunoglobulin frameworks or scaffolds include, but are not limited to, Adnectin (fibronectin) (Compound Therapeutics, Inc., Waltham, MA), ankyrin (Molecular Partners AG, Zurich, Switzerland), domain antibodies (Domantis, Ltd (Cambridge, MA) and Ablynx nv (Zwijnaarde, Belgium)), lipocalins (Anticalins) (Pieris Proteolab AG, Freising, Germany), small modular immunopharmaceuticals (Trubion Pharmaceuticals Inc., Seattle, WA), maxybodies (Avidia, Inc. (Mountain View, CA)), protein A (Affibody AG, Sweden), and affilin (γ-crystallin or ubiquitin) (Scil Proteins GmbH, Halle, Germany), protein epitope mimetics (Polyphor Ltd, Allschwil, Switzerland).

[0387] (i) Fibronectin scaffold

[0388] The fibronectin scaffold is preferably based on a type III fibronectin domain (e.g., the tenth module of type III fibronectin (10Fn3 domain)). The type III fibronectin domain has 7 or 8 β-strands distributed between two β-sheets, which themselves wrap around each other to form a protein core and further contain loops (similar to CDRs) that connect the β-strands to each other and are exposed to the solvent. There are at least three such loops at each edge of the β-sheet sandwich, where the edge is the boundary of the protein perpendicular to the direction of the β-strands (US 6,818,418).

[0389] These fibronectin-based scaffolds are not immunoglobulins, although the overall fold is closely related to the folding of the smallest functional antibody fragment, the heavy chain variable region, which contains the complete antigen recognition unit in camel and llama IgG. Due to this structure, non-immunoglobulin antibodies mimic antigen binding properties similar to antibodies in terms of properties and affinity. These scaffolds can be used for in vitro loop randomization and shuffling strategies, which are similar to the in vivo affinity maturation process of antibodies. These fibronectin-based molecules can be used as scaffolds, wherein the loop regions of the molecule can be replaced with the CDRs of the present disclosure using standard cloning techniques.

[0390] (ii) Ankyrin-molecular partner

[0391] This technology is based on the use of proteins with ankyrin-derived repeat modules as scaffolds to host variable regions that can be used to bind to different targets. The ankyrin repeat module is a 33-amino acid polypeptide composed of two antiparallel α-helices and a β-turn. The binding of the variable regions is primarily optimized using ribosome display.

[0392] (iii) Maxi-Avimers-Avidia

[0393] Avimers are derived from proteins containing natural A domains, such as LRP-1. These domains are naturally used for protein-protein interactions, and in humans, more than 250 proteins are structurally based on A domains. Avimers are composed of many different "A-domain" monomers (2-10) connected by amino acid linkers. Avimers capable of binding to target antigens can be produced using methods described in, for example, US2004 / 0175756; US2005 / 0053973; US2005 / 0048512; and US2006 / 0008844.

[0394] (vi) Protein A-Affibody

[0395] The affinity ligand is a simple small protein consisting of a three-helix bundle based on a scaffold from one of the IgG binding domains of protein A. Protein A is a surface protein from the bacterium Staphylococcus aureus. This scaffold domain consists of 58 amino acids, 13 of which are randomized to generate a large number of ligand variants. Libraries (see, for example, US 5,831,012). The molecule mimics antibodies, with a molecular weight of 6kDa, compared to 150kDa for antibodies. Despite its small size, The binding site of the molecule is similar to that of an antibody.

[0396] (v)Anticalins-Pieris

[0397] These products are developed by Pieris ProteoLab AG. They are derived from lipocalins, a widespread group of small, robust proteins that are often involved in the physiological transport or storage of chemically sensitive or insoluble compounds. Several naturally occurring lipocalins are found in human tissues and fluids. The protein architecture is reminiscent of immunoglobulins, with hypervariable loops atop a rigid framework. However, in contrast to antibodies or their recombinant fragments, lipocalins consist of a single polypeptide chain of 160 to 180 amino acid residues, only slightly larger than a single immunoglobulin domain. The set of four loops that form the binding pocket exhibits significant structural plasticity and tolerates a variety of side chains. Therefore, the binding site can be reshaped in a proprietary process to recognize target molecules of varying shapes with high affinity and specificity.

[0398] One protein of the lipocalin family, the biletriene binding protein (BBP) of Pieris brassicae, has been used to develop anticalins by mutagenesis of the set of four loops. An example of a patent application describing "anticalins" is WO 1999 / 16873.

[0399] (vi) Affilin-Scil Proteins

[0400] AFFILIN TM The molecules are small non-immunoglobulin proteins designed for specific affinity to proteins and small molecules. New AFFILINs can be selected very quickly from two libraries TM The libraries are composed of molecules, each of which is based on a different human scaffold protein.

[0401] AFFILIN TMThe molecule does not show any structural homology to immunoglobulin proteins. Scil Proteins utilizes two AFFILIN TM The scaffolds are γ-crystalline, a human structural eye lens protein, and the other is a protein from the "ubiquitin" superfamily. Both human scaffolds are very small, exhibit high-temperature stability, and are nearly resistant to pH changes and denaturants. This high stability is primarily due to the protein's extended beta-sheet structure. Examples of γ-crystalline-derived proteins are described in WO 2001 / 004144, and examples of "ubiquitin-like" proteins are described in WO 2004 / 106368.

[0402] (vii) Protein epitope mimetics (PEMs)

[0403] PEMs are medium-sized cyclic peptide-like molecules (MW 1-2 kDa) that mimic the β-hairpin secondary structure of proteins, which is the main secondary structure involved in protein-protein interactions.

[0404] Grafting antigen-binding domains into alternative frameworks or scaffolds

[0405] A variety of antibody / immunoglobulin frameworks or scaffolds can be employed, so long as the resulting polypeptide comprises at least one binding region that specifically binds to ActRIIB. Such frameworks or scaffolds include the five major idiotypes of human immunoglobulins or fragments thereof (such as those disclosed elsewhere herein), and include immunoglobulins from other animal species, preferably with humanized aspects. In this regard, single heavy chain antibodies, such as those identified in camelids, are of particular interest. Those skilled in the art will continue to explore and develop new frameworks, scaffolds, and fragments.

[0406] In one aspect, the compositions of the present disclosure may comprise non-immunoglobulin-based antibodies using non-immunoglobulin scaffolds onto which the CDRs of the disclosed antibodies may be grafted. Known or future non-immunoglobulin frameworks and scaffolds may be employed, as long as they comprise a binding region specific for the target protein shown in SEQ ID NO: 181 (preferably, a ligand binding domain thereof as shown in SEQ ID NO: 182). Such compounds are referred to herein as "polypeptides comprising a target-specific binding region". Examples of non-immunoglobulin frameworks are further described in the following sections (Camelidae Antibodies and Non-Antibody Scaffolds).

[0407] Framework or Fc engineering

[0408] The engineered antibodies included in the compositions of the present disclosure include those in which V H and / or VL In some embodiments, the framework residues within the antibody are modified, for example to improve the properties of the antibody. Typically, such framework modifications are performed to reduce the immunogenicity of the antibody. For example, one method is to "backmutate" one or more framework residues to the corresponding germline sequence. More specifically, an antibody that has undergone somatic mutation may contain framework residues that are different from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequence with the germline sequence from which the antibody is derived. In order to restore the framework region sequence to its germline configuration, the somatic mutation can be "backmutated" to the germline sequence by, for example, site-directed mutagenesis or PCR-mediated mutagenesis. Such "backmutated" antibodies may also be included in the compositions of the present disclosure.

[0409] Another type of framework modification involves mutating one or more residues within the framework region or even within one or more CDR regions to remove T cell epitopes, thereby reducing the potential immunogenicity of the antibody. This approach is also known as "deimmunization" and is described in further detail in US2003 / 0153043.

[0410] As a supplement or alternative to modifications made in the framework or CDR regions, the antibodies of the present disclosure can be engineered to include modifications in the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular toxicity. In addition, the antibodies included in the compositions of the present disclosure can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or can be modified to alter its glycosylation to again alter one or more functional properties of the antibody. Each of these embodiments is described in further detail below. The numbering of residues in the Fc region is that of the EU index of Kabat.

[0411] In one embodiment, the hinge region of CH1 is modified such that the number of cysteine ​​residues in the hinge region is altered, e.g., increased or decreased. This approach is further described in US Pat. No. 5,677,425. The number of cysteine ​​residues in the hinge region of CH1 is altered to, for example, facilitate assembly of the light and heavy chains or to increase or decrease antibody stability.

[0412] In another embodiment, the Fc hinge region of an antibody is mutated to reduce the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcal protein A (SpA) binding relative to native-Fc hinge domain SpA binding. This approach is further described in US 6,165,745.

[0413] In another embodiment, the antibody is modified to extend its biological half-life. Various methods are possible. For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in US Pat. No. 6,277,375. Alternatively, to extend biological half-life, the antibody can be modified within the CH1 or CL region to include salvage receptor binding epitopes derived from two loops of the CH2 domain of the Fc region of IgG, as described in US Pat. Nos. 5,869,046 and 6,121,022.

[0414] In other embodiments, the Fc region is changed by replacing at least one amino acid residue with a different amino acid residue to change the effector function of the antibody. For example, one or more amino acids can be replaced with different amino acid residues so that the antibody has a changed affinity for the effector ligand, but retains the antigen binding ability of the parent antibody. The effector ligand to which its affinity is changed can be, for example, an Fc receptor or the C1 component of complement. This method is further described in detail in US5,624,821 and US5,648,260 by Winter et al. Specifically, residues 234 and 235 can be mutated. Specifically, these mutations can be to become alanine. Therefore, in one embodiment, the antibody included in the composition of the present disclosure has a mutation at one or both of amino acids 234 and 235 in the Fc region. In another embodiment, one or both of amino acids 234 and 235 can be substituted with alanine. Both amino acids 234 and 235 are substituted with alanine, resulting in reduced ADCC activity.

[0415] In another embodiment, one or more amino acids selected from the amino acid residues of the antibody can be replaced with a different amino acid residue such that the antibody has altered C1q binding and / or reduced or abolished complement dependent cytotoxicity (CDC). This approach is described in further detail in US 6,194,551.

[0416] In another embodiment, one or more amino acid residues of the antibody are altered to thereby alter the ability of the antibody to fix complement. This approach is further described in WO94 / 29351.

[0417] In another embodiment, the Fc region of the antibody is modified by modifying one or more amino acids to enhance the ability of the antibody to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or to enhance the affinity of the antibody for Fcγ receptors. This approach is further described in WO00 / 42072. In addition, the binding sites of FcγRI, FcγRII, FcγRIII, and FcRn on human IgG1 have been mapped, and variants with improved binding have been described (see Shields, RL et al., 2001 J. Biol. Chen. 276: 6591-6604).

[0418] In another embodiment, the glycosylation of the antibody contained in the composition of the present disclosure is modified. For example, an aglycoslated antibody can be prepared (i.e., the antibody lacks glycosylation). Glycosylation can be changed to, for example, increase the affinity of the antibody for the antigen. Such carbohydrate modifications can be achieved by, for example, changing one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made that result in the elimination of one or more variable region framework glycosylation sites, thereby eliminating glycosylation at the site. Such glycosylation can enhance the affinity of the antibody for the antigen. Such methods are further described in detail in U.S. Patent Nos. 5,714,350 and 6,350,861 to Co et al.

[0419] Additionally or alternatively, antibodies with altered glycosylation patterns may be used, such as hypofucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GsNac structures. Such altered glycosylation patterns have been shown to enhance the ADCC ability of antibodies. Such carbohydrate modifications can be achieved, for example, by expressing the antibody in a h...

Claims

1. Use of an ActRII receptor antibody in the preparation of a medicament for treating and / or preventing heart failure, wherein the ActRII receptor antibody comprises: The heavy chain variable region CDR1 shown in SEQ ID NO: 9; the heavy chain variable region CDR2 shown in SEQ ID NO: 23; the heavy chain variable region CDR3 shown in SEQ ID NO: 37; the light chain variable region CDR1 shown in SEQ ID NO: 51; the light chain variable region CDR2 shown in SEQ ID NO: 65; and the light chain variable region CDR3 shown in SEQ ID NO:

79.

2. The use according to claim 1, wherein the heart failure is caused by or is associated with at least one of the following: valvular heart disease, coronary artery disease, hypertension, diabetes, aging, arrhythmia, peripartum cardiomyopathy, stress cardiomyopathy, toxic or infectious agents, hereditary cardiomyopathy or idiopathic dilated cardiomyopathy.

3. The use according to claim 1, wherein the heart failure is heart failure with reduced ejection fraction.

4. The use according to claim 1, wherein the heart failure is heart failure with preserved ejection fraction.

5. The use according to claim 2, wherein the valvular heart disease is aortic stenosis.

6. The use according to claim 5, wherein the aortic stenosis is accompanied by frailty and / or sarcopenia.

7. The use according to claim 2, wherein the peripartum cardiomyopathy occurs during late pregnancy or within 6 months after delivery.

8. The use according to claim 2, wherein the stress cardiomyopathy occurs following psychological, pathological or physical stress.

9. The use according to any one of claims 1-8, wherein the K of the ActRII receptor antibody is D is 2 pM.

10. The use according to any one of claims 1-8, wherein the ActRII receptor antibody binds to ActRIIB with an affinity that is 10 times or greater than its affinity for binding to ActRIIA.

11. The use according to any one of claims 1-8, wherein the ActRII receptor antibody comprises: The heavy chain sequence shown in SEQ ID NO: 146, or a sequence having at least 95% sequence identity thereto, and the light chain sequence shown in SEQ ID NO: 141, or a sequence having at least 95% sequence identity thereto.

12. The use according to claim 11, wherein the ActRII receptor antibody has altered effector function by mutation of the Fc region.

13. The use according to claim 11, wherein the ActRII receptor antibody is encoded by pBW522 deposited with the deposit number DSM22873 or pBW524 deposited with the deposit number DSM22874.

14. The use according to claim 11, wherein the ActRII receptor antibody is bimagrumab.

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