WO2010125003A1 - Compositions and methods for increasing muscle growth - Google Patents

Compositions and methods for increasing muscle growth Download PDF

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Publication number
WO2010125003A1
WO2010125003A1 PCT/EP2010/055458 EP2010055458W WO2010125003A1 WO 2010125003 A1 WO2010125003 A1 WO 2010125003A1 EP 2010055458 W EP2010055458 W EP 2010055458W WO 2010125003 A1 WO2010125003 A1 WO 2010125003A1
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Prior art keywords
seq
antibody
variable region
chain variable
light chain
Prior art date
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PCT/EP2010/055458
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English (en)
French (fr)
Inventor
Catrin Berger
Tanja Herrmann
Chris Lu
Kelly-Ann Sheppard
Estelle Trifilieff
Stefanie Urlinger
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Novartis Ag
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Priority to BRPI1014522A priority Critical patent/BRPI1014522B8/pt
Priority to AU2010243697A priority patent/AU2010243697B2/en
Application filed by Novartis Ag filed Critical Novartis Ag
Priority to SI201031603T priority patent/SI2424895T1/en
Priority to LTEP10714332.3T priority patent/LT2424895T/lt
Priority to CN201080018660.2A priority patent/CN102753578B/zh
Priority to RS20171263A priority patent/RS56661B1/sr
Priority to MX2011011352A priority patent/MX2011011352A/es
Priority to JP2012507695A priority patent/JP5766179B2/ja
Priority to NO10714332A priority patent/NO2424895T3/no
Priority to NZ595235A priority patent/NZ595235A/xx
Priority to SG2011064359A priority patent/SG174273A1/en
Priority to CA2758290A priority patent/CA2758290C/en
Priority to PL10714332T priority patent/PL2424895T3/pl
Priority to EP10714332.3A priority patent/EP2424895B1/en
Priority to UAA201112627A priority patent/UA102437C2/ru
Priority to DK10714332.3T priority patent/DK2424895T3/en
Priority to MA34357A priority patent/MA33279B1/fr
Priority to EA201101571A priority patent/EA027071B1/ru
Priority to KR1020117028110A priority patent/KR101836501B1/ko
Priority to ES10714332.3T priority patent/ES2655877T3/es
Publication of WO2010125003A1 publication Critical patent/WO2010125003A1/en
Priority to ZA2011/06538A priority patent/ZA201106538B/en
Priority to TN2011000463A priority patent/TN2011000463A1/fr
Priority to IL215168A priority patent/IL215168A0/en
Priority to HRP20171870TT priority patent/HRP20171870T1/hr
Priority to CY20171101282T priority patent/CY1119664T1/el

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    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
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    • C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
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    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
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    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • This invention is in the field of anti-Activin receptor HB (ActRIIB) antibodies.
  • it relates to the use of said antibodies for treating muscle disorders, such as muscle wasting due to disease or disuse.
  • Activins are dimeric growth and differentiation factors which belong to the transforming growth factor-beta (TGF-beta) superfamily of structurally related signaling proteins. Activins signal through a heterodimeric complex of receptor serine kinases which include at least two type I (I and IB) and two type II (II and HB, aka ACVR2A and ACVR2B) receptors. These receptors are all transmembrane proteins, composed of a ligand-binding extracellular domain with cysteine-rich region, a transmembrane domain, and a cytoplasmic domain with predicted serine/threonine specificity.
  • TGF-beta transforming growth factor-beta
  • Type I receptors are essential for signalling while type II receptors are required for binding ligands and for expression of type I receptors.
  • Type I and II receptors form a stable complex after ligand binding resulting in the phosphorylation of type I receptors by type II receptors.
  • the activin receptor II B (ActRIIB) is a receptor for myostatin.
  • the interation between myostatin and this receptor regulates the inhibition of skeletal muscle differentiation via the Smad-dependent pathway.
  • the invention provides an anti-ActRIIB antibody, or a functional protein comprising an antigen-binding portion of said antibody.
  • the ActRIIB is human ActRIIB.
  • the polypeptide sequence of human ActRIIB is recited in SEQ ID NO: 181 (AAC64515.1, GL3769443).
  • the antibody or functional protein is from a mammal, having an origin such as human or camelid.
  • the antibody may be a chimeric, human or a humanized antibody.
  • the anti-ActRIIB antibody is characterized as having antigen-binding region that is specific for the target protein ActRIIB and binds to ActRIIB or a fragment of ActRIIB.
  • the antibody is suitable for use in therapy.
  • the antibodies according to the invention are ActRIIB antagonists with no or low agonistic activity.
  • the antibody or functional fragment binds the target protein ActRIIB and decreases the binding of myostatin to ActRIIB to a basal level.
  • the antibody or functional fragment reduces the amount of myostatin that binds to ActRIIB.
  • the antibody or functional fragment completely prevents myostatin from binding to ActRIIB.
  • the antibody or functional fragment inhibits Smad activation.
  • the antibody or functional fragment inhibits activin receptor type HB mediated myostatin-induced inhibition of skeletal differentiation via the Smad-dependent pathway.
  • the binding may be determined by one or more assays that can be used to measure an activity which is either antagonism or agonism by the antibody.
  • the assays measure at least one of the effects of the antibody on ActRIIB that include: inhibition of myostatin binding to
  • the invention provides antibodies that specifically bind to the myostatin binding region (i.e. ligand binding domain) of ActRIIB.
  • This ligand binding domain consists of amino acids 19-134 of SEQ ID NO: 181 and has been assigned SEQ ID NO: 182 herein.
  • the antibodies bind to ActRIIB with a K D of 10OnM or less, 1OnM or less, InM or less.
  • the antibodies of the invention bind to ActRIIB with an affinity of lOOpM or less (i.e.lOOpM, 5OpM, lOpM, IpM or less).
  • the antibodies of the invention bind to ActRIIB with an affinity of between 10 and 2OpM.
  • the antibodies of the invention do not cross-react with an ActRIIB related protein, and more particularly do not cross-react with human ActRIIA (NP_001607.1, GL4501897).
  • the antibodies of the invention preferentially bind to ActRIIB rather than ActRIIA. In one embodiment, the antibodies of the invention bind to ActRIIB with 5-fold greater affinity than they bind to ActRIIA, more preferably 10-fold, still more preferably 50-fold, still more preferably 100-fold. In one embodiment, the antibodies of the invention bind to ActRIIA with an affinity of lOOpM or more (i.e.250pM, 50OpM, InM, 5nM or more).
  • the antibodies of the invention are of the IgG2 isotype.
  • the antibodies of the invention are of the IgGl isotype. In a further embodiment, the antibodies of the invention are of the IgGl isotype and have an altered effector function through mutation of the Fc region. In one embodiment, said altered effector function is reduced ADCC and CDC activity. In one embodiment, said altered effector function is silenced ADCC and CDC activity.
  • the antibodies according to the invention are fully human or humanized IgGl antibodies with no antibody dependent cellular cytotoxicity (ADCC) activity or CDC activity and bind to a region of ActRIIB consisting of amino acids 19-134 of SEQ ID NO:181.
  • ADCC antibody dependent cellular cytotoxicity
  • the antibodies according to the invention are fully human or humanized IgGl antibodies with reduced antibody dependent cellular cytotoxicity (ADCC) activity or CDC activity and bind to a region of ActRIIB consisting of amino acids 19-134 of SEQ ID NO:181.
  • the present invention relates to isolated antibodies, particularly human or humanized antibodies that inhibit myostatin binding to ActRIIB and activate skeletal muscle differentiation in vitro and in vivo.
  • the antibodies of the invention are derived from particular heavy and light chain sequences and/or comprise particular structural features such as CDR regions comprising particular amino acid sequences.
  • the invention provides isolated antibodies, methods of making such antibodies, immunoconjugates and multivalent or multispecific molecules comprising such antibodies and pharmaceutical compositions containing the antibodies, immunoconjugates or bispecific molecules of the invention.
  • the invention also relates to methods of using the antibodies to inhibit, i.e. antagonize, function of ActRIIB in order to inhibit Smad activation and thereby induce skeletal muscle differentiation, for example, resulting in the treatment of a pathological disorder.
  • the pathological disorder may be a musculoskeletal disease or disorder, such as muscle atrophy.
  • muscle atrophy There are many causes of muscle atrophy, including as a result of treatment with a glucocorticoid such as Cortisol, dexamethasone, betamethasone, prednisone, methylprednisolone, or prednisolone.
  • the muscle atrophy can also be a result of denervation due to nerve trauma or a result of degenerative, metabolic, or inflammatory neuropathy (e.g., Guillian-Barre syndrome, peripheral neuropathy, or exposure to environmental toxins or drugs).
  • the muscle atrophy can be a result of myopathy, such as myotonia; a congential myopathy, including nemalene myopathy, multi/minicore myopathy and myotubular (centronuclear) myopathy; mitochondrial myopathy; familial periodic paralysis; inflammatory myopathy; metabolic myopathy, such as caused by a glycogen or lipid storage disease; dermatomyositisis; polymyositis; inclusion body myositis; myositis ossificans; rhabdomyolysis and myoglobinurias.
  • myopathy such as myotonia; a congential myopathy, including nemalene myopathy, multi/minicore myopathy and myotubular (centronuclear) myopathy; mitochondrial myopathy; familial periodic paralysis; inflammatory myopathy; metabolic myopathy, such as caused by a glycogen or lipid storage disease; dermatomyositisis; polymyositis; inclusion body myositis; my
  • the myopathy may be caused by a muscular dystrophy syndrome, such as Duchenne, Becker, myotonic, fascioscapulohumeral, Emery-Dreifuss, oculopharyngeal, scapulohumeral, limb girdle, Fukuyama, a congenital muscular dystrophy, or hereditary distal myopathy.
  • the musculoskeletal disease can also be osteoporosis, a bone fracture, short stature, or dwarfism.
  • the muscle atrophy can be a result of an adult motor neuron disease, infantile spinal muscular atrophy, amyotrophic lateral sclerosis, juvenile spinal muscular atrophy, autoimmune motor neuropathy with multifocal conductor block, paralysis due to stroke or spinal cord injury, skeletal immobilization due to trauma, prolonged bed rest, voluntary inactivity, involuntary inactivity, metabolic stress or nutritional insufficiency, cancer, AIDS, fasting, a thyroid gland disorder, diabetes, benign congenital hypotonia, central core disease, burn injury, chronic obstructive pulmonary disease, liver diseases (examples such as fibrosis, cirrhosis), sepsis, renal failure, congestive heart failure, ageing, space travel or time spent in a zero gravity environment.
  • adult motor neuron disease infantile spinal muscular atrophy, amyotrophic lateral sclerosis, juvenile spinal muscular atrophy, autoimmune motor neuropathy with multifocal conductor block, paralysis due to stroke or spinal cord injury, skeletal immobilization due to trauma, prolonged
  • age-related conditions examples include, sarcopenia, skin atrophy, muscle wasting, brain atrophy, atherosclerosis, arteriosclerosis, pulmonary emphysema, osteoporosis, osteoarthritis, immunologic incompetence, high blood pressure, dementia, Huntington's disease, Alzheimer's disease, cataracts, age-related macular degeneration, prostate cancer, stroke, diminished life expectancy, frailty, memory loss, wrinkles, impaired kidney function, and age- related hearing loss; metabolic disorders, including Type II Diabetes, Metabolic Syndrome, hyperglycemia, and obesity.
  • Other conditions that may be treated with the antibodies of the invention include acute and/or chronic renal disease or failure, liver fibrosis or cirrhosis, cancer such as breast cancer, Parkinson's Disease; conditions associated with neuronal death, such as ALS, brain atrophy, or dementia and anemia.
  • Further conditions include cachexia, cachexia associated with a rheumatoid arthritis and cachexia associated with cancer.
  • the antibodies of the invention may be used to treat liver, kidney, pulmonary fibrosis and cancers examplified by but not restricted to rhabdomyosarcomas, bone-loss inducing cancers, hepatocellular carcinomas, gastrointestinal cancers.
  • the prevention may be complete, e.g., the total absence of an age-related condition or metabolic disorder.
  • the prevention may also be partial, such that the likelihood of the occurrence of the age-related condition or metabolic disorder in a subject is less likely to occur than had the subject not received an antibody of the present invention.
  • immune response refers to the action of, for example, lymphocytes, antigen presenting cells, phagocytic cells, granulocytes, and soluble macromolecules produced by the above cells or the liver (including antibodies, cytokines, and complement) that results in selective damage to, destruction of, or elimination from the human body of invading pathogens, cells or tissues infected with pathogens, cancerous cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues.
  • a “signal transduction pathway” or “signaling activity” refers to a biochemical causal relationship generally initiated by a protein-protein interaction such as binding of a growth factor to a receptor, resulting in transmission of a signal from one portion of a cell to another portion of a cell.
  • the transmission involves specific phosphorylation of one or more tyrosine, serine, or threonine residues on one or more proteins in the series of reactions causing signal transduction.
  • Penultimate processes typically include nuclear events, resulting in a change in gene expression.
  • ActRIIB or Act HB receptor refers to human ActRIIB as defined in SEQ ID NO: 181 (AAC64515.1, GL3769443).
  • Research grade polyclonal and monoclonal anti-ActRIIB antibodies are known in the art, such as those made by R&D Systems ® , MN, USA.
  • Therapeutic anti-ActRIIB antibodies have not previously been described. Of course, antibodies could be raised against ActRIIB from other species and used to treat pathological conditions in those species.
  • antibody as referred to herein includes whole antibodies and any antigen binding fragment (i.e. "antigen-binding portion") or single chains thereof.
  • a naturally occurring "antibody” is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds.
  • Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as V H ) and a heavy chain constant region.
  • the heavy chain constant region is comprised of three domains, CHl, CH2 and CH3.
  • Each light chain is comprised of a light chain variable region (abbreviated herein as V L ) and a light chain constant region.
  • the light chain constant region is comprised of one domain, C L .
  • V H and V L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR).
  • CDR complementarity determining regions
  • FR framework regions
  • Each V H and V L is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FRl, CDRl, FR2, CDR2, FR3, CDR3, FR4.
  • the variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
  • the constant regions of the antibodies may 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 (CIq) of the classical complement system.
  • antigen-binding portion of an antibody refers to full length 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.
  • binding fragments encompassed within the term "antigen-binding portion" of an antibody include a Fab fragment, a monovalent fragment consisting of the V L , V H , C L and CHl domains; a F(ab) 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the V H and CHl domains; a Fv fragment consisting of the V L and V H domains of a single arm of an antibody; a dAb fragment (Ward et al., 1989 Nature 341:544-546), which consists of a V H domain; and an isolated complementarity determining region (CDR).
  • Fab fragment a monovalent fragment consisting of the V L , V H , C L and CHl domains
  • F(ab) 2 fragment a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region
  • a Fd fragment consisting of the V H and
  • the two domains of the Fv fragment, V L and V H are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the V L and V H regions pair to form monovalent molecules (known as 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).
  • single chain Fv single chain Fv
  • Such single chain antibodies are also intended to be encompassed within the term "antigen-binding region" of an antibody.
  • an "isolated antibody”, as used herein, refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g. an isolated antibody that specifically binds ActRIIB is substantially free of antibodies that specifically bind antigens other than ActRIIB).
  • An isolated antibody that specifically binds ActRIIB may, however, have cross- reactivity to other antigens, such as ActRIIB molecules from other species.
  • an isolated antibody may be substantially free of other cellular material and/or chemicals.
  • monoclonal antibody or “monoclonal antibody composition” as used herein refer 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.
  • human antibody is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region also is derived from such human sequences, e.g. human germline sequences, or mutated versions of human germline sequences or antibody containing consensus framework sequences derived from human framework sequences analysis, for example, as described in Knappik, et al. (2000. J MoI Biol 296, 57-86).
  • human antibodies of the invention may include amino acid residues not encoded by human sequences (e.g. mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo).
  • human antibody as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
  • human monoclonal antibody refers to antibodies displaying a single binding specificity which have variable regions in which both the framework and CDR regions are derived from human sequences.
  • the human monoclonal antibodies are produced by a hybridoma which includes a B cell obtained from a transgenic nonhuman animal, e.g. a transgenic mouse, having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell.
  • recombinant human antibody includes all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from an animal (e.g. a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom, antibodies isolated from a host cell transformed to express the human antibody, e.g. from a transfectoma, antibodies isolated from a recombinant, combinatorial human antibody library, and antibodies prepared, expressed, created or isolated by any other means that involve splicing of all or a portion of a human immunoglobulin gene, sequences 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.
  • such recombinant human antibodies can be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the V H and V L regions of the recombinant antibodies are sequences that, while derived from and related to human germline V H and V L sequences, may not naturally exist within the human antibody germline repertoire in vivo.
  • isotype refers to the antibody class (e.g. IgM, IgE, IgG such as IgGl or IgG2) that is provided by the heavy chain constant region genes.
  • the phrases “an antibody recognizing an antigen” and “an antibody specific for an antigen” are used interchangeably herein with the term “an antibody which binds specifically to an antigen”.
  • an antibody that "specifically binds to ActRIIB polypeptide” is intended to refer to an antibody that binds to human ActRIIB polypeptide with a K D of a 10OnM or less, 1OnM or less, InM or less.
  • An antibody that "cross-reacts with an antigen other than ActRIIB” is intended to refer to an antibody that binds that antigen with a K D of 10 x lO "9 M or less, 5 x 10 ⁇ 9 M or less, or 2 x 10 "9 M or less.
  • An antibody that "does not cross-react with a particular antigen” is intended to refer to an antibody that binds to that antigen, with a K D of 1.5 x 10 8 M or greater, or a K D of 5-10 x 10 "8 M, or 1 x 10 "7 M or greater.
  • K D may be determined using a biosensor system, such as a Biacore ® system, or Solution Equilibrium Titration.
  • the term "antagonist antibody” is intended to refer to an antibody that inhibits ActRIIB induced signaling activity in the presence of myostatin.
  • an assay to detect this include inhibition of myostatin induced signalling (for instance 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 (for instance by a creatine kinase assay).
  • the antibodies inhibit myostatin induced signalling as measured in a Smad dependent reporter gene assay at an IC50 of 1OnM or less, InM or less, or lOOpM or less.
  • an antibody with "no agonistic activity” is intended to refer to an antibody that does not significantly increase ActRIIB mediated signaling activity in the absence of myostatin in a cell-based assay, such as inihibition of myostatin induced signalling (for instance by a Smad dependent reporter gene assay), inhibition of myostatin induced Smad phosphorylation (P-Smad
  • K assoc or "K a ", as used herein, is intended to refer to the association rate of a particular antibody-antigen interaction
  • K d15 or “K d”
  • K D is intended to refer to the dissociation rate of a particular antibody-antigen interaction
  • K D is intended to refer to the dissociation constant, which is obtained from the ratio of K d to K a (i.e. K d /K a ) and is expressed as a molar concentration (M).
  • K D values for antibodies can be determined using methods well established in the art.
  • a method for determining the K D of an antibody is by using surface plasmon resonance, such as the biosensor system of Biacore ® , 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).
  • surface plasmon resonance such as the biosensor system of Biacore ®
  • SET Solution Equilibrium Titration
  • Affinity refers to the strength of interaction between antibody and antigen at single antigenic sites. Within each antigenic site, the variable region of the antibody “arm” interacts through weak non-covalent forces with antigen at numerous sites; the more interactions, the stronger the affinity.
  • the term "Avidity” refers to an informative measure of the overall stability or strength of the antibody-antigen complex. It is controlled by three major factors: antibody epitope affinity; the valency of both the antigen and antibody; and the structural arrangement of the interacting parts. Ultimately these factors define the specificity of the antibody, that is, the likelihood that the particular antibody is binding to a precise antigen epitope.
  • ADCC antibody dependent cellular cytotoxicity activity refers to human B cell depleting activity. ADCC activity can be measured by the human B cell depleting assays known in the art. In order to get a higher avidity probe, a dimeric conjugate (two molecules of an antibody protein coupled to a FACS marker) can be constructed, thus making low affinity interactions (such as with the germline antibody) more readily detected by FACS. In addition, another means to increase the avidity of antigen binding involves generating dimers, trimers or multimers of any of the constructs described herein of the anti-ActRIIB antibodies.
  • Such multimers may be generated through covalent binding between individual modules, for example, by imitating the natural C-to-N-terminus binding or by imitating antibody dimers that are held together through their constant regions.
  • the bonds engineered into the Fc/Fc interface may be covalent or non- covalent.
  • dimerizing or multimerizing partners other than Fc can be used in ActRIIB hybrids to create such higher order structures. For example, it is possible to use multimerizing domains such as the trimerizing domain described in WO2004/039841 or pentamerizing domain described in WO98/18943.
  • the term "selectivity" for an antibody refers to an antibody that binds to a certain target polypeptide but not to closely related polypeptides.
  • the term “high affinity” for an antibody refers to an antibody having a K D of InM or less for a target antigen.
  • the term “subject” includes any human or nonhuman animal.
  • the term “nonhuman animal” includes all vertebrates, e.g. mammals and non-mammals, such as nonhuman primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc.
  • the term, "optimized" means that a nucleotide sequence has been altered to encode an amino acid sequence using codons that are preferred in the production cell or organism, generally a eukaryotic cell, for example, a cell of Pichia, a cell of Trichoderma, a Chinese Hamster Ovary cell (CHO) or a human cell.
  • 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 known as the "parental" sequence.
  • the optimized sequences herein have been engineered to have codons that are preferred in CHO mammalian cells, however optimized expression of these sequences in other eukaryotic cells is also envisioned herein.
  • the amino acid sequences encoded by optimized nucleotide sequences are also referred to as optimized.
  • Standard assays to evaluate the binding ability of the antibodies toward ActRIIB of various species are known in the art, including for example, ELISAs, western blots and RIAs. Suitable assays are described in detail in the Examples.
  • the binding affinity of the antibodies also can be assessed by standard assays known in the art, such as by Biacore analysis or Solution
  • an antibody that "inhibits" one or more of these ActRIIB functional properties e.g. biochemical, immunochemical, cellular, physiological or other biological activities, or the like
  • an antibody that inhibits ActRIIB activity effects such a statistically significant decrease by at least 10% of the measured parameter, by at least 50%, 80% or 90%, and in certain embodiments an antibody of the invention may inhibit greater than 95%, 98% or 99% of ActRIIB functional activity.
  • cross-block means 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.
  • 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 ActRIIB, and therefore whether it can be said to cross-block according to the invention, can be determined using standard competition binding assays.
  • One suitable assay involves the use of the Biacore technology (e.g. by using a BIAcore instrument (Biacore, Uppsala, Sweden)), which can measure the extent of interactions using surface plasmon resonance technology.
  • Another assay for measuring cross-blocking uses an ELISA-based approach.
  • a further assay uses FACS analysis, wherein competition of various antibodies for binding to ActRIIB expressing cells is tested (such as described in the Examples).
  • a cross-blocking antibody or other binding agent binds to ActRIIB in the described BIAcore cross-blocking assay such that the recorded binding of the combination (mixture) of the antibodies or binding agents is between 80% and 0.1% (e.g. 80% to 4%) of the maximum theoretical binding, specifically between 75% and 0.1% (e.g. 75% to 4%) of the maximum theoretical binding, and more specifically between 70% and 0.1% (e.g. 70% to 4%), and more specifically between 65% and 0.1% (e.g. 65% to 4%) of maximum theoretical binding (as defined above) of the two antibodies or binding agents in combination.
  • An antibody is defined as cross-blocking an anti- ActRIIB antibody of the invention in an ELISA assay, if the test antibody is able to cause a reduction of anti-ActRIIB antibody binding to ActRIIB of between 60% and 100%, specifically between 70% and 100%, and more specifically between 80% and 100%, when compared to the positive control wells (i.e. the same anti-ActRIIB antibody and ActRIIB, but no "test" cross-blocking antibody).
  • Examples of cross blocking antibodies as cited herein are MOR08159 and MOR08213.
  • the invention provides antibodies that cross block MOR08159 or MOR08213 for binding to ActRIIB.
  • Antibodies of the invention include the human recombinant antibodies, isolated and structurally characterized, as described in the Examples.
  • the V H amino acid sequences of isolated antibodies of the invention are shown in SEQ ID NOs: 99-112.
  • the V L amino acid sequences of isolated antibodies of the invention are shown in SEQ ID NOs: 85-98 respectively.
  • Examples of preferred full length heavy chain amino acid sequences of antibodies of the invention are shown in SEQ ID NOs: 146-150 and 156-160.
  • Examples of preferred full length light chain amino acid sequences of antibodies of the invention are shown in SEQ ID NOs: 141-145 and 151-155 respectively.
  • antibodies of the invention include amino acids that have been mutated by amino acid deletion, insertion or substitution, yet have at least 60, 70, 80, 90, 95, 97 or 99 percent identity in the CDR regions with the CDR regions depicted in the sequences described above. In some embodiments, it includes mutant amino acid sequences wherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated by amino acid deletion, insertion or substitution in the CDR regions when compared with the CDR regions depicted in the sequence described above.
  • variable heavy chain parental nucleotide sequences are shown in SEQ ID NOs: 127-140.
  • Variable light chain parental nucleotide sequences are shown in SEQ ID NOs: 113-126.
  • Full length light chain nucleotide sequences optimized for expression in a mammalian cell are shown in SEQ ID NOs: 161-165 and 171-175.
  • Full length heavy chain nucleotide sequences optimized for expression in a mammalian cell are shown in SEQ ID NOs: 166-170 and 176-180.
  • Other antibodies of the invention include amino acids or nucleic acids that have been mutated, yet have at least 60 or more (i.e. 80, 90, 95, 97, 99 or more) percent identity to the sequences described above. In some embodiments, it includes mutant amino acid sequences wherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated by amino acid deletion, insertion or substitution in the variable regions when compared with the variable regions depicted in the sequence described above.
  • V H , V L , full length light chain, and full length heavy chain sequences can be "mixed and matched" to create other anti-ActRIIB binding molecules of the invention.
  • ActRIIB binding of such "mixed and matched" antibodies can be tested using the binding assays described above and in the Examples (e.g. ELISAs).
  • ELISAs e.g. ELISAs
  • a 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.
  • a V L sequence from a particular V H /V L pairing should be replaced with a structurally similar V L sequence.
  • a full length light chain sequence from a particular full length heavy chain / full length light chain pairing should be replaced with a structurally similar full length light chain sequence.
  • the invention provides an isolated recombinant anti-ActRIIB 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 NOs: 99-112; and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 85-98.
  • the invention provides:
  • an isolated recombinant anti-ActRIIB antibody having: a full length heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:99-112; and a full length light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:85-98, or
  • an isolated recombinant anti-ActRIIB antibody having a full length heavy chain encoded by a nucleotide sequence that has been optimized for expression in the cell of a mammalian selected from the group consisting of SEQ ID NOs: 127- 140, and a full length light chain encoded by a nucleotide sequence that has been optimized for expression in the cell of a mammalian selected from the group consisting of SEQ ID NOs:113-126, or
  • the amino acid sequences of the V H CDRIS of the antibodies are shown in SEQ ID NOs: 1-14.
  • the amino acid sequences of the V H CDR2S of the antibodies are shown in SEQ ID NOs: 15-28.
  • amino acid sequences of the V H CDR3S of the antibodies are shown in SEQ ID NOs: 29-42.
  • amino acid sequences of the V L CDRIS of the antibodies are shown in SEQ ID NOs: 43-56.
  • amino acid sequences of the V L CDR2S of the antibodies are shown in SEQ ID NOs: 57-70.
  • the amino acid sequences of the V L CDR3S of the antibodies are shown in SEQ ID NOs: 71-84.
  • the CDR regions are delineated using the Kabat system (Kabat, E. A., et al., 1991 Sequences of
  • V H CDRl, 2 and 3 sequences and V L CDRl, 2 and 3 sequences can be "mixed and matched" (i.e. CDRs from different antibodies can be mixed and matched, each antibody containing a V H CDRl, 2 and 3 and a V L CDRl, 2 and 3 create other anti-ActRIIB binding molecules of the invention.
  • ActRIIB binding of such "mixed and matched" antibodies can be tested using the binding assays described above and in the Examples (e.g. ELISAs).
  • V H CDR sequences When V H CDR sequences are mixed and matched, the CDRl, CDR2 and/or CDR3 sequence from a particular V H sequence should be replaced with a structurally similar CDR sequence(s).
  • V L CDR sequences when V L CDR sequences are mixed and matched, the CDRl, CDR2 and/or CDR3 sequence from a particular V L sequence should be replaced with a structurally similar CDR sequence(s).
  • novel V H and V L sequences can be created by substituting one or more V H and/or V L CDR region sequences with structurally similar sequences from the CDR sequences shown herein for monoclonal antibodies of the present invention.
  • An isolated recombinant anti-ActRIIB antibody, or antigen binding region thereof has: a heavy chain variable region CDRl comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-14; a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 15-28; a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 29-42; a light chain variable region CDRl comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-56; a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 57-70; and a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 71-84.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 1
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 2 a heavy chain variable region CDR2 of SEQ ID NO: 16; a heavy chain variable region CDR3 of SEQ ID NO: 30; a light chain variable region CDRl of SEQ ID NO: 44; a light chain variable region CDR2 of SEQ ID NO: 58; and a light chain variable region CDR3 of SEQ ID NO: 72.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 3; a heavy chain variable region CDR2 of SEQ ID NO: 17; a heavy chain variable region CDR3 of SEQ ID NO: 31; a light chain variable region CDRl of SEQ ID NO: 45; a light chain variable region CDR2 of SEQ ID NO: 59; and a light chain variable region CDR3 of SEQ ID NO: 73.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 4; a heavy chain variable region CDR2 of SEQ ID NO: 18; a heavy chain variable region CDR3 of SEQ ID NO: 32; a light chain variable region CDRl of SEQ ID NO: 46; a light chain variable region CDR2 of SEQ ID NO: 60; and a light chain variable region CDR3 of SEQ ID NO: 74.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 5; a heavy chain variable region CDR2 of SEQ ID NO: 19; a heavy chain variable region CDR3 of SEQ ID NO: 33; a light chain variable region CDRl of SEQ ID NO: 47; a light chain variable region CDR2 of SEQ ID NO: 61; and a light chain variable region CDR3 of SEQ ID NO: 75.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 6; a heavy chain variable region CDR2 of SEQ ID NO: 20; a heavy chain variable region CDR3 of SEQ ID NO: 34; a light chain variable region CDRl of SEQ ID NO: 48; a light chain variable region CDR2 of SEQ ID NO: 62; and a light chain variable region CDR3 of SEQ ID NO: 76.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 7; a heavy chain variable region CDR2 of SEQ ID NO: 21; a heavy chain variable region CDR3 of SEQ ID NO: 35; a light chain variable region CDRl of SEQ ID NO: 49; a light chain variable region CDR2 of SEQ ID NO: 63; and a light chain variable region CDR3 of SEQ ID NO: 77.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 8; a heavy chain variable region CDR2 of SEQ ID NO: 22; a heavy chain variable region
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 9; a heavy chain variable region CDR2 of SEQ ID NO: 23; a heavy chain variable region CDR3 of SEQ ID NO: 37; a light chain variable region CDRl of SEQ ID NO: 51; a light chain variable region CDR2 of SEQ ID NO: 65; and a light chain variable region CDR3 of SEQ ID NO: 79.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 10; a heavy chain variable region CDR2 of SEQ ID NO: 24; a heavy chain variable region CDR3 of SEQ ID NO: 38; a light chain variable region CDRl of SEQ ID NO: 52; a light chain variable region CDR2 of SEQ ID NO: 66; and a light chain variable region CDR3 of SEQ ID NO: 80.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 11; a heavy chain variable region CDR2 of SEQ ID NO: 25; a heavy chain variable region CDR3 of SEQ ID NO: 39; a light chain variable region CDRl of SEQ ID NO: 53; a light chain variable region CDR2 of SEQ ID NO: 67; and a light chain variable region CDR3 of SEQ ID NO: 81.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 12; a heavy chain variable region CDR2 of SEQ ID NO: 26; a heavy chain variable region CDR3 of SEQ ID NO: 40; a light chain variable region CDRl of SEQ ID NO: 54; a light chain variable region CDR2 of SEQ ID NO: 68; and a light chain variable region CDR3 of SEQ ID NO: 82.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 13; a heavy chain variable region CDR2 of SEQ ID NO: 27; a heavy chain variable region CDR3 of SEQ ID NO: 41; a light chain variable region CDRl of SEQ ID NO: 55; a light chain variable region CDR2 of SEQ ID NO: 69; and a light chain variable region CDR3 of SEQ ID NO: 83.
  • the antibody comprises: a heavy chain variable region CDRl of SEQ ID NO: 14; a heavy chain variable region CDR2 of SEQ ID NO: 28; a heavy chain variable region CDR3 of SEQ ID NO: 42; a light chain variable region CDRl of SEQ ID NO: 56; a light chain variable region CDR2 of SEQ ID NO: 70; and a light chain variable region CDR3 of SEQ ID NO: 84.
  • the invention provides an antibody comprising: (a) the variable heavy chain sequence of SEQ ID NO: 85 and variable light chain sequence of SEQ ID NO: 99; (b) the variable heavy chain sequence of SEQ ID NO: 86 and variable light chain sequence of SEQ ID NO: 100; (c) the variable heavy chain sequence of SEQ ID NO: 87 and variable light chain sequence of SEQ ID NO: 101; (d) the variable heavy chain sequence of SEQ ID NO: 88 and variable light chain sequence of SEQ ID NO: 102; (e) the variable heavy chain sequence of SEQ ID NO: 89 and variable light chain sequence of SEQ ID NO: 103; (f) the variable heavy chain sequence of SEQ ID NO: 90 and variable light chain sequence of SEQ ID NO: 104; (g) the variable heavy chain sequence of SEQ ID NO: 91 and variable light chain sequence of SEQ ID NO: 105; (h) the variable heavy chain sequence of SEQ ID NO: 92 and variable light chain sequence of SEQ ID NO: 106; (i) the variable heavy chain sequence
  • the invention provides an antibody comprising: (a) the heavy chain sequence of SEQ ID NO: 146 and light chain sequence of SEQ ID NO: 141; (b) the heavy chain sequence of SEQ ID NO: 147 and light chain sequence of SEQ ID NO: 142; (c) the heavy chain sequence of SEQ ID NO: 148 and light chain sequence of SEQ ID NO: 143; (d) the heavy chain sequence of SEQ ID NO: 149 and light chain sequence of SEQ ID NO: 144; (e) the heavy chain sequence of SEQ ID NO: 150 and light chain sequence of SEQ ID NO: 145; (f) the heavy chain sequence of SEQ ID NO: 156 and light chain sequence of SEQ ID NO: 151; (g) the heavy chain sequence of SEQ ID NO: 157 and light chain sequence of SEQ ID NO: 152; (h) the heavy chain sequence of SEQ ID NO: 158 and light chain sequence of SEQ ID NO: 153; (i) the heavy chain sequence of SEQ ID NO: 159 and light chain sequence of
  • a human antibody comprises heavy or light chain variable regions or full length heavy or light chains that are "the product of” or “derived from” a particular germline sequence if the variable regions or full length chains of the antibody are obtained from a system that uses human germline immunoglobulin genes.
  • Such systems include immunizing a transgenic mouse carrying human immunoglobulin genes with the antigen of interest or screening a human immunoglobulin gene library displayed on phage with the antigen of interest.
  • a human antibody that is "the product of” or “derived from” a human germline immunoglobulin sequence can be identified as such by comparing the amino acid sequence of the human antibody to the amino acid sequences of human germline immunoglobulins and selecting the human germline immunoglobulin sequence that is closest in sequence (i.e. greatest % identity) to the sequence of the human antibody.
  • a human antibody that is "the product of” or “derived from” a particular human germline immunoglobulin sequence may contain amino acid differences as compared to the germline sequence, due to, for example, naturally occurring somatic mutations or intentional introduction of site-directed mutation.
  • a selected human antibody typically is at least 90% identical in amino acids sequence to an amino acid sequence encoded by a human germline immunoglobulin gene and contains amino acid residues that identify the human antibody as being human when compared to the germline immunoglobulin amino acid sequences of other species (e.g. murine germline sequences).
  • a human antibody may be at least 80%, 90%, or at least 95%, or even at least 96%, 97%, 98%, or 99% identical in amino acid sequence to the amino acid sequence encoded by the germline immunoglobulin gene.
  • a human antibody derived from a particular human germline sequence will display no more than 10 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene.
  • the human antibody may display no more than 5, or even no more than 4, 3, 2, or 1 amino acid difference from the amino acid sequence encoded by the germline immunoglobulin gene.
  • the antibody of the invention is that encoded by pBW522 or pBW524 (deposited at DSMZ, Inhoffenstr. 7B, D-38124 Braunschweig, Germany on 18 August 2009 under deposit numbers DSM22873 and DSM22874, respectively).
  • an antibody of the invention has full length heavy and light chain amino acid sequences; full length heavy and light chain nucleotide sequences, variable region heavy and light chain nucleotide sequences, or variable region heavy and light chain amino acid sequences that are homologous to the amino acid and nucleotide sequences of the antibodies described herein, and wherein the antibodies retain the desired functional properties of the anti- ActRIIB antibodies of the invention.
  • the invention provides an isolated recombinant anti-ActRIIB antibody (or a functional protein comprising an antigen binding portion thereof) comprising 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 NOs: 99-112; 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 NOs: 85-98; and the antibody exhibits at least one of the following functional properties: (i) it inhibits myo statin binding in vitro or in vivo and/or (ii) decreases inhibition of muscle differentiation through the Smad-dependent pathway.
  • 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
  • the invention provides an isolated recombinant anti-ActRIIB antibody, (or a functional protein comprising an antigen binding portion thereof) comprising 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 NOs: 146-150 and 156-160; 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 NOs: 141-145 and 151-155; and the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo and/or (ii) decreases inhibition of muscle differentiation through the Smad-dependent pathway.
  • the invention provides an isolated recombinant anti-ActRIIB antibody (or a functional protein comprising an antigen binding portion thereof), comprising 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 NOs: 166-170 and 176-180; 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 NOs: 161-165 and 171-175; and the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in viv
  • the antibody may exhibit one or more, two or more, or three of the functional properties discussed above.
  • the antibody can be, for example, a human antibody, a humanized antibody or a chimeric antibody.
  • the antibody is a fully human IgGl antibody.
  • V H and/or V L amino acid sequences may be 80%, 90%, 95%, 96%,
  • V H and/or V L amino acid sequences may be identical except an amino acid substitution in no more than 1, 2, 3, 4 or 5 amino acid position.
  • An antibody having V H and V L regions having high i.e.
  • 98 respectively can be obtained by mutagenesis (e.g. site-directed or PCR-mediated mutagenesis) of nucleic acid molecules SEQ ID NOs: 127-140 and 113-126 respectively, followed by testing of the encoded altered antibody for retained function (i.e. the functions set forth above) using the functional assays described herein.
  • mutagenesis e.g. site-directed or PCR-mediated mutagenesis
  • the full length heavy chain and/or full length light chain amino acid sequences may be 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences set forth above.
  • An antibody having a full length heavy chain and full length light chain having high (i.e. 80% or greater) identity to the full length heavy chains of any of SEQ ID NOs: 146-150 and 156- 160 and full length light chains of any of SEQ ID NOs: 141-145 and 151-155 respectively, can be obtained by mutagenesis (e.g.
  • the full length heavy chain and/or full length light chain nucleotide sequences may be 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences set forth above.
  • variable regions of heavy chain and/or light chain nucleotide sequences may be 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences set forth above.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described below.
  • 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 PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. MoI, 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 either 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.
  • an antibody of the invention has a heavy chain variable region comprising CDRl, CDR2, and CDR3 sequences and a light chain variable region comprising CDRl, CDR2, and CDR3 sequences, wherein one or more of these CDR sequences have specified amino acid sequences based on the antibodies described herein or conservative modifications thereof, and wherein the antibodies retain the desired functional properties of the anti-ActRIIB antibodies of the invention.
  • the invention provides an isolated recombinant anti-ActRIIB antibody, or a functional protein comprising an antigen binding portion thereof, consisting of a heavy chain variable region comprising CDRl, CDR2, and CDR3 sequences and a light chain variable region comprising CDRl, CDR2, and CDR3 sequences, wherein: the heavy chain variable region CDRl amino acid sequences are selected from the group consisting of SEQ ID NOs: 1-14, and conservative modifications thereof; the heavy chain variable region CDR2 amino acid sequences are selected from the group consisting of SEQ ID NOs: 15-28, and conservative modifications thereof; the heavy chain variable region CDR3 amino acid sequences are selected from the group consisting of SEQ ID NOs: 29-42, and conservative modifications thereof; the light chain variable regions CDRl amino acid sequences are selected from the group consisting of SEQ ID NOs: 43-56, and conservative modifications thereof; the light chain variable regions CDR2 amino acid sequences are selected from the group consisting of SEQ ID NOs: 57-70,
  • the antibody exhibits at least one of the following functional properties: (i) it inhibits myostatin binding in vitro or in vivo and/or (ii) decreases inhibition of muscle differentiation through the Smad-dependent pathway.
  • the antibody may exhibit one or both of the functional properties listed above.
  • Such antibodies can be, for example, human antibodies, humanized antibodies or chimeric antibodies.
  • an antibody of the invention optimized for expression in a mammalian cell has a full length heavy chain sequence and a full length light chain sequence, wherein one or more of these sequences have specified amino acid sequences based on the antibodies described herein or conservative modifications thereof, and wherein the antibodies retain the desired functional properties of the anti-ActRIIB antibodies of the invention.
  • the invention provides an isolated monoclonal anti-ActRIIB antibody optimized for expression in a mammalian cell consisting of a full length heavy chain and a full length light chain wherein: the full length heavy chain has amino acid sequences selected from the group of SEQ ID NOs: 146- 150 and 156-160, and conservative modifications thereof; and the full length light chain has amino acid sequences selected from the group of SEQ ID NOs: 141-145 and 151-155, 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 and/or (ii) decreases inhibition of muscle differentiation through the Smad-dependent pathway.
  • the antibody may exhibit one or both of the functional properties listed above.
  • Such antibodies can be, for example, human antibodies, humanized antibodies or chimeric antibodies.
  • conservative sequence modifications is intended to refer 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 an antibody of the invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis.
  • Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain.
  • Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g. lysine, arginine, histidine), acidic side chains (e.g. aspartic acid, glutamic acid), uncharged polar side chains (e.g. glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g.
  • beta-branched side chains e.g. threonine, valine, isoleucine
  • aromatic side chains e.g. tyrosine, phenylalanine, tryptophan, histidine.
  • Antibodies that bind to the same epitope as anti-ActRIIB antibodies of the invention provides antibodies that bind to the same epitope as the various specific anti-ActRIIB antibodies of the invention described herein. All the antibodies described in the Examples that are capable of blocking myostatin binding to ActRIIB bind the same epitope in ActRIIB with high affinity, said epitope being comprised between amino acids 19-134 of SEQ ID NO:181. Additional antibodies can therefore be identified based on their ability to cross-compete (e.g. to competitively inhibit the binding of, in a statistically significant manner) with other antibodies of the invention in standard ActRIIB binding assays.
  • test antibody to inhibit the binding of antibodies of the present invention to human ActRIIB demonstrates that the test antibody can compete with that antibody for binding to human ActRIIB; such an antibody may, according to non-limiting theory, bind to the same or a related (e.g. a structurally similar or spatially proximal) epitope on human ActRIIB as the antibody with which it competes.
  • the antibody that binds to the same epitope on human ActRIIB as the antibodies of the present invention is a human recombinant antibody.
  • human recombinant antibodies can be prepared and isolated as described in the Examples.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 85, and the variable light chain sequence recited in SEQ ID NO: 99.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 86, and the variable light chain sequence recited in SEQ ID NO: 100.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 87, and the variable light chain sequence recited in SEQ ID NO: 101.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 88, and the variable light chain sequence recited in SEQ ID NO: 102.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 89, and the variable light chain sequence recited in SEQ ID NO: 103.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 90, and the variable light chain sequence recited in SEQ ID NO: 104.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 91, and the variable light chain sequence recited in SEQ ID NO: 105.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 92, and the variable light chain sequence recited in SEQ ID NO: 106.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 93, and the variable light chain sequence recited in SEQ ID NO: 107.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 94, and the variable light chain sequence recited in SEQ ID NO: 108.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 95, and the variable light chain sequence recited in SEQ ID NO: 109.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 96, and the variable light chain sequence recited in SEQ ID NO: 110.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 97, and the variable light chain sequence recited in SEQ ID NO: 111.
  • the invention provides an antibody that binds to an epitope recognised by an antibody having the variable heavy chain sequence recited in SEQ ID NO: 98, and the variable light chain sequence recited in SEQ ID NO: 112. Following more detailed epitope mapping experiments, the binding regions of preferred antibodies of the invention have been more clearly defined.
  • the invention provides an antibody that binds to an epitope comprising amino acids 78-83 of SEQ ID NO: 181 (WLDDFN - SEQ ID NO:188).
  • the invention also provides an antibody that binds to an epitope comprising amino acids 76-84 of SEQ ID NO: 181 (GCWLDDFNC - SEQ ID NO: 186).
  • the invention also provides an antibody that binds to an epitope comprising amino acids 75-85 of SEQ ID NO: 181 (KGCWLDDFNCY - SEQ ID NO:190).
  • the invention also provides an antibody that binds to an epitope comprising amino acids 52-56 of SEQ ID NO: 181 (EQDKR - SEQ ID NO:189).
  • the invention also provides an antibody that binds to an epitope comprising amino acids 49-63 of SEQ ID NO: 181 (CEGEQDKRLHCYASW - SEQ ID NO:187).
  • the invention also provides antibodies that bind to epitopes consisting of these sequences or epitopes comprising combinations of these epitope regions.
  • the invention also provides 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).
  • An antibody of the invention further can be prepared using an antibody having one or more of the V H and/or V L sequences shown herein as starting material to engineer a modified antibody, which modified antibody may have altered properties from the starting antibody.
  • An antibody can be engineered by modifying one or more residues within one or both variable regions (i.e. V H and/or V L ), for example within one or more CDR regions and/or within one or more framework regions. Additionally or alternatively, an antibody can be engineered by modifying residues within the constant region(s), for example to alter the effector function(s) of the antibody.
  • CDR grafting One type of variable region engineering that can be performed is CDR grafting.
  • Antibodies interact with target antigens predominantly through amino acid residues that are located in the six heavy and light chain complementarity determining regions (CDRs). For this reason, the amino acid sequences within CDRs are more diverse between individual antibodies than sequences outside of CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express recombinant antibodies that mimic the properties of specific naturally occurring antibodies by constructing expression vectors that include CDR sequences from the 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.
  • another embodiment of the invention pertains to an isolated monoclonal anti- ActRIIB antibody, or a functional protein comprising an antigen binding portion thereof, comprising a heavy chain variable region comprising CDRl sequences having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-14; CDR2 sequences having an amino acid sequence selected from the group consisting of SEQ ID NOs: 15-28; CDR3 sequences having an amino acid sequence selected from the group consisting of SEQ ID NOs: 29-42, respectively; and a light chain variable region having CDRl sequences having an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-56; CDR2 sequences having an amino acid sequence selected from the group consisting of SEQ ID NOs: 57-70; and CDR3 sequences consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 71-84, respectively.
  • such antibodies contain the V H and V L CDR sequences of monoclonal antibodies, yet may contain different framework sequence
  • Such framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences.
  • germline DNA sequences for 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, E. A., et al., [supra]; Tomlinson, I. M., et al., 1992 J. fol. Biol. 227:776-798; and Cox, J. P. L. et al., 1994 Eur. J Immunol. 24:827-836.
  • framework sequences for use in the antibodies of the invention are those that are structurally similar to the framework sequences used by selected antibodies of the invention, e.g. consensus sequences and/or framework sequences used by monoclonal antibodies of the invention.
  • the V H CDRl, 2 and 3 sequences, and the V L CDRl, 2 and 3 sequences can be grafted onto framework regions that have the identical sequence as that found in the germline immunoglobulin gene from which the framework sequence derive, or the CDR sequences can be grafted onto framework regions that contain one or more mutations as compared to the germline sequences.
  • variable region modification is to mutate amino acid residues within the V H and/or V L CDRl, CDR2 and/or CDR3 regions to thereby improve one or more binding properties (e.g.
  • affinity maturation affinity of the antibody of interest
  • Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce the mutation(s) and the effect on antibody binding, or other functional property of interest, can be evaluated in in vitro or in vivo assays as described herein and provided in the Examples. Conservative modifications (as discussed above) can be introduced.
  • the mutations may be amino acid substitutions, additions or deletions. Moreover, typically no more than one, two, three, four or five residues within a CDR region are altered.
  • the invention provides isolated anti-ActRIIB monoclonal antibodies, or a functional protein comprising an antigen binding portion thereof, consisting of a heavy chain variable region having: a V H CDRl region consisting of an amino acid sequence selected from the group having SEQ ID NOs: 1-14 or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions as compared to SEQ ID NOs: 1-14; a V H CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 15-28, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions as compared to SEQ ID NOs: 15-28; a V H CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 29-42, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions as compared to SEQ ID NOs: 29-42; a V H CDRl region consist
  • antibody/immunoglobulin frameworks or scaffolds can be employed so long as the resulting polypeptide includes at least one binding region which specifically binds to ActRIIB.
  • Such frameworks or scaffolds include the 5 main idiotypes of human immunoglobulins, or fragments thereof (such as those disclosed elsewhere herein), and include immunoglobulins of other animal species, preferably having humanized aspects. Single heavy- chain antibodies such as those identified in camelids are of particular interest in this regard. Novel frameworks, scaffolds and fragments continue to be discovered and developed by those skilled in the art.
  • the invention pertains to generating non-immunoglobulin based antibodies using non-immunoglobulin scaffolds onto which CDRs of the invention can 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 of SEQ ID NO: 181 (preferably, the ligand binding domain thereof as shown in SEQ ID NO: 182).
  • Such compounds are known herein as "polypeptides comprising a target-specific binding region”. Examples of non- immunoglobulin framework are further described in the sections below (camelid antibodies and non-antibody scaffold).
  • Antibody proteins obtained from members of the camel and dromedary family (Camelus bactrianus and Camelus dromaderius) including new world members such as llama species
  • Lama paccos, Lama glama and Lama vicugna have been characterized with respect to size, structural complexity and antigenicity for human subjects.
  • Certain IgG antibodies from this family of mammals as found in nature lack light chains, and are thus structurally distinct from the typical four chain quaternary structure having two heavy and two light chains, for antibodies from other animals (see WO94/04678).
  • a region of the camelid antibody which is the small single variable domain identified as V HH can be obtained by genetic engineering to yield a small protein having high affinity for a target, resulting in a low molecular weight antibody-derived protein known as a "camelid nanobody” (see US5,759,808; 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.
  • the camelid nanobody has a molecular weight approximately one-tenth that of a human IgG molecule, and the protein has a physical diameter of only a few nanometers.
  • One consequence of the small size is the ability of camelid nanobodies to bind to antigenic sites that are functionally invisible to larger antibody proteins, i.e. camelid nanobodies are useful as reagents detect antigens that are otherwise cryptic using classical immunological techniques, and as possible therapeutic agents.
  • a camelid nanobody can inhibit as a result of binding to a specific site in a groove or narrow cleft of a target protein, and hence can serve in a capacity that more closely resembles the function of a classical low molecular weight drug than that of a classical antibody.
  • the low molecular weight and compact size further result in camelid nanobodies being extremely thermostable, stable to extreme pH and to proteolytic digestion, and poorly antigenic.
  • camelid nanobodies readily move from the circulatory system into tissues, and even cross the blood-brain barrier and can treat disorders that affect nervous tissue. Nanobodies can further facilitate drug transport across the blood brain barrier (see US2004/0161738). These features combined with the low antigenicity to humans indicate great therapeutic potential. Further, these molecules can be fully expressed in prokaryotic cells such as E. coli and are expressed as fusion proteins with bacteriophage and are functional.
  • a feature of the present invention is a camelid antibody or nanobody having high affinity for ActRIIB.
  • the camelid antibody or nanobody is naturally produced in the camelid animal, i.e. is produced by the camelid following immunization with ActRIIB or a peptide fragment thereof, using techniques described herein for other antibodies.
  • the anti-ActRIIB camelid nanobody is engineered, i.e. produced by selection for example from a library of phage displaying appropriately mutagenized camelid nanobody proteins using panning procedures with ActRIIB as a target as described in the examples herein.
  • Engineered nanobodies can further be customized by genetic engineering to have a half life in a recipient subject of from 45 minutes to two weeks.
  • the camelid antibody or nanobody is obtained by grafting the CDRs sequences of the heavy or light chain of the human antibodies of the invention into nanobody or single domain antibody framework sequences, as described for example in WO94/04678.
  • Non-immunoglobulin frameworks or scaffolds include, but are not limited to, Adnectins (fibronectin) (Compound Therapeutics, Inc., Waltham, MA), ankyrin (Molecular Partners AG,
  • the fibronectin scaffolds are based preferably on fibronectin type III domain (e.g. the tenth module of the fibronectin type III (10 Fn3 domain)).
  • the fibronectin type III domain has 7 or 8 beta strands which are distributed between two beta sheets, which themselves pack against each other to form the core of the protein, and further containing loops (analogous to CDRs) which connect the beta strands to each other and are solvent exposed. There are at least three such loops at each edge of the beta sheet sandwich, where the edge is the boundary of the protein perpendicular to the direction of the beta strands (US 6,818,418).
  • fibronectin-based scaffolds are not an immunoglobulin, although the overall fold is closely related to that of the smallest functional antibody fragment, the variable region of the heavy chain, which comprises the entire antigen recognition unit in camel and llama IgG. Because of this structure, the non-immunoglobulin antibody mimics antigen binding properties that are similar in nature and affinity to those of antibodies.
  • These scaffolds can be used in a loop randomization and shuffling strategy in vitro that is similar to the process of affinity maturation of antibodies in vivo.
  • These fibronectin-based molecules can be used as scaffolds where the loop regions of the molecule can be replaced with CDRs of the invention using standard cloning techniques.
  • ankyrin - Molecular Partners The technology is based on using proteins with ankyrin derived repeat modules as scaffolds for bearing variable regions which can be used for binding to different targets.
  • the ankyrin repeat module is a 33 amino acid polypeptide consisting of two anti-parallel ⁇ -helices and a ⁇ -turn. Binding of the variable regions is mostly optimized by using ribosome display.
  • Avimers are derived from natural A-domain containing protein such as LRP-I. These domains are used by nature for protein-protein interactions and in human over 250 proteins are structurally based on A-domains. Avimers consist of a number of different "A-domain” monomers (2-10) linked via amino acid linkers. Avimers can be created that can bind to the target antigen using the methodology described in, for example, US2004/0175756; US2005/0053973; US2005/0048512; and US2006/0008844. (vi) Protein A - Affibody
  • Affibody® affinity ligands are small, simple proteins composed of a three-helix bundle based on the scaffold of 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 Affibody® libraries with a large number of ligand variants (See e.g. US 5,831,012).
  • Affibody® molecules mimic antibodies, they have a molecular weight of 6 kDa, compared to the molecular weight of antibodies, which is 150 kDa. In spite of its small size, the binding site of Affibody® molecules is similar to that of an antibody.
  • Anticalins® are products developed by the company Pieris ProteoLab AG. They are derived from lipocalins, a widespread group of small and robust proteins that are usually involved in the physiological transport or storage of chemically sensitive or insoluble compounds. Several natural lipocalins occur in human tissues or body liquids.
  • lipocalins are composed of a single polypeptide chain with 160 to 180 amino acid residues, being just marginally bigger than a single immunoglobulin domain.
  • the set of four loops which makes up the binding pocket, shows pronounced structural plasticity and tolerates a variety of side chains.
  • the binding site can thus be reshaped in a proprietary process in order to recognize prescribed target molecules of different shape with high affinity and specificity.
  • One protein of lipocalin family the bilin-binding protein (BBP) of Pieris brassicae has been used to develop anticalins by mutagenizing the set of four loops.
  • BBP bilin-binding protein
  • One example of a patent application describing "anticalins" is WO1999/16873.
  • AffilinTM molecules are small non-immunoglobulin proteins which are designed for specific affinities towards proteins and small molecules. New AffilinTM molecules can be very quickly selected from two libraries, each of which is based on a different human derived scaffold protein.
  • AffilinTM molecules do not show any structural homology to immunoglobulin proteins.
  • Scil Proteins employs two AffilinTM scaffolds, one of which is gamma crystalline, a human structural eye lens protein and the other is "ubiquitin” superfamily proteins. Both human scaffolds are very small, show high temperature stability and are almost resistant to pH changes and denaturing agents. This high stability is mainly due to the expanded beta sheet structure of the proteins. Examples of gamma crystalline derived proteins are described in WO2001/004144 and examples of "ubiqui tin-like” proteins are described in WO2004/106368. (vii) Protein Epitope Mimetics (PEM)
  • PEM are medium- sized, cyclic, peptide-like molecules (MW l-2kDa) mimicking beta-hairpin secondary structures of proteins, the major secondary structure involved in protein -protein interactions.
  • Framework or Fc engineering Engineered antibodies of the invention include those in which modifications have been made to framework residues within V H and/or V L , e.g. to improve the properties of the antibody. Typically such framework modifications are made to decrease the immunogenicity of the antibody. For example, one approach 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 differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequences to the germline sequences from which the antibody is derived.
  • the somatic mutations can be "backmutated” to the germline sequence by, for example, site-directed mutagenesis or PCR- mediated mutagenesis.
  • Such "backmutated” antibodies are also intended to be encompassed by the invention.
  • 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 to thereby reduce the potential immunogenicity of the antibody. This approach is also referred to as "deimmunization" and is described in further detail in US2003/0153043.
  • antibodies of the invention may be engineered to include modifications within 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 cytotoxicity.
  • an antibody of the invention may be chemically modified (e.g. one or more chemical moieties can be attached to the antibody) or be modified to alter its glycosylation, again to alter one or more functional properties of the antibody.
  • the hinge region of CHl is modified such that the number of cysteine residues in the hinge region is altered, e.g. increased or decreased. This approach is described further in US5,677,425.
  • the number of cysteine residues in the hinge region of CHl is altered to, for example, facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.
  • the Fc hinge region of an antibody is mutated to decrease 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 Staphylococcyl protein A (SpA) binding relative to native Fc-hinge domain SpA binding.
  • SpA Staphylococcyl protein A
  • the antibody is modified to increase its biological half-life.
  • Various approaches are possible. For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in US6,277,375.
  • the antibody can be altered within the CHl or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG, as described in US5,869,046 and US6,121,022.
  • the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector functions of the antibody.
  • one or more amino acids can be replaced with a different amino acid residue such that the antibody has an altered affinity for an effector ligand but retains the antigen -binding ability of the parent antibody.
  • the effector ligand to which affinity is altered can be, for example, an Fc receptor or the Cl component of complement.
  • residues 234 and 235 may be mutated. In particular, these mutations may be to alanine.
  • the antibody of the invention has a mutation in the Fc region at one or both of amino acids 234 and 235.
  • one or both of amino acids 234 and 235 may be substituted to alanine. Substitution of both amino acids 234 and 235 to alanine results in a reduced ADCC activity.
  • one or more amino acids selected from amino acid residues can be replaced with a different amino acid residue such that the antibody has altered CIq binding and/or reduced or abolished complement dependent cytotoxicity (CDC). This approach is described in further detail in US6, 194,551.
  • one or more amino acid residues are altered to thereby alter the ability of the antibody to fix complement.
  • the Fc region is modified to increase the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and/or to increase the affinity of the antibody for an Fc ⁇ receptor by modifying one or more amino acids.
  • ADCC antibody dependent cellular cytotoxicity
  • WO00/42072 the binding sites on human IgGl for Fc ⁇ Rl, Fc ⁇ RII, Fc ⁇ RIII and FcRn have been mapped and variants with improved binding have been described (see Shields, R.L. et al., 2001 J. Biol. Chen. 276:6591-6604).
  • the glycosylation of an antibody is modified.
  • an aglycoslated antibody can be made (i.e. the antibody lacks glycosylation).
  • Glycosylation can be altered to, for example, increase the affinity of the antibody for the antigen.
  • carbohydrate modifications can be accomplished by; for example, altering one or more sites of glycosylation within the antibody sequence.
  • one or more amino acid substitutions can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site.
  • Such aglycosylation may increase the affinity of the antibody for antigen.
  • an antibody can be made that has an altered type of glycosylation, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNac structures.
  • altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies.
  • carbohydrate modifications can be accomplished by, for example, expressing the antibody in a host cell with altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells in which to express recombinant antibodies of the invention to thereby produce an antibody with altered glycosylation.
  • the antibodies of the invention are produced by recombinant expression in a cell line which exhibit hypofucosylation pattern, for example, a mammalian cell line with deficient expression of the FUT8 gene encoding fucosyltransferase.
  • WO03/035835 describes a variant CHO cell line, Lecl3 cells, with reduced ability to attach fucose to Asn(297)- linked carbohydrates, also resulting in hypofucosylation of antibodies expressed in that host cell (see also Shields, R.L. et al., 2002 J.
  • WO99/54342 describes cell lines engineered to express glycoprotein-modifying glycosyl transferases (e.g. beta(l,4)-N acetylglucosaminyltransferase III (GnTIII)) such that antibodies expressed in the engineered cell lines exhibit increased bisecting GIcN ac structures which results in increased ADCC activity of the antibodies (see also Umana et al., 1999 Nat. Biotech. 17:176-180).
  • glycoprotein-modifying glycosyl transferases e.g. beta(l,4)-N acetylglucosaminyltransferase III (GnTIII)
  • the antibodies of the invention can be produced in a yeast or a filamentous fungi engineered for mammalian-like glycosylation pattern, and capable of producing antibodies lacking fucose as glycosylation pattern (see for example EP1297172B1).
  • An antibody can be pegylated to, for example, increase the biological (e.g. serum) half-life of the antibody.
  • the antibody, or fragment thereof typically is reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment.
  • PEG polyethylene glycol
  • the pegylation can be carried out by an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer).
  • polyethylene glycol is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (Cl-ClO) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide.
  • the antibody to be pegylated is an aglycosylated antibody. Methods for pegylating proteins are known in the art and can be applied to the antibodies of the invention (see for example, EP0154316 and EP0401384).
  • Another modification of the antibodies that is contemplated by the invention is a conjugate or a protein fusion of at least the antigen-binding region of the antibody of the invention to serum protein, such as human serum albumin or a fragment thereof to increase half- life of the resulting molecule (see, for example, EP0322094).
  • serum protein such as human serum albumin or a fragment thereof to increase half- life of the resulting molecule
  • Another possibility is a fusion of at least the antigen -binding region of the antibody of the invention to proteins capable of binding to serum proteins, such human serum albumin to increase half life of the resulting molecule (see, for example, EP0486525).
  • the anti-ActRIIB antibodies having V H and V L sequences or full length heavy and light chain sequences shown herein can be used to create new anti-ActRIIB antibodies by modifying full length heavy chain and/or light chain sequences, V H and/or V L sequences, or the constant region(s) attached thereto.
  • the structural features of an anti-ActRIIB antibody of the invention are used to create structurally related anti- ActRIIB antibodies that retain at least one functional property of the antibodies of the invention, such as binding to human ActRIIB but also inhibit one or more functional properties of ActRIIB (for example, the inhibition of Smad activation).
  • one or more CDR regions of the antibodies of the present invention, or mutations thereof can be combined recombinantly with known framework regions and/or other CDRs to create additional, recombinantly-engineered, anti-ActRIIB antibodies of the invention, as discussed above.
  • the starting material for the engineering method is one or more of the V H and/or V L sequences provided herein, or one or more CDR regions thereof.
  • To create the engineered antibody it is not necessary to actually prepare (i.e. express as a protein) an antibody having one or more of the V H and/or V L sequences provided herein, or one or more CDR regions thereof. Rather, the information contained in the sequence(s) is used as the starting material to create a "second generation" sequence(s) derived from the original sequence(s) and then the "second generation" sequence(s) is prepared and expressed as a protein.
  • the invention provides a method for preparing an anti-ActRIIB antibody consisting of: a heavy chain variable region antibody sequence having a CDRl sequence selected from the group consisting of SEQ ID NOs: 1-14, a CDR2 sequence selected from the group consisting of SEQ ID NOs: 15-28 and/or a CDR3 sequence selected from the group consisting of SEQ ID NOs: 29-42; and a light chain variable region antibody sequence having a CDRl sequence selected from the group consisting of SEQ ID NOs: 43-56, a CDR2 sequence selected from the group consisting of SEQ ID NOs: 57-70 and/or a CDR3 sequence selected from the group consisting of SEQ ID NOs: 71-84; altering at least one amino acid residue within the heavy chain variable region antibody sequence and/or the light chain variable region antibody sequence to create at least one altered antibody sequence; and expressing the altered antibody sequence as a protein.
  • the invention provides a method for preparing an anti-ActRIIB antibody optimized for expression in a mammalian cell consisting of: a full length heavy chain antibody sequence having a sequence selected from the group of SEQ ID NOs: 146- 150 and 156-160; and a full length light chain antibody sequence having a sequence selected from the group of SEQ ID NOs: 141-145 and 151-155; altering at least one amino acid residue within the full length heavy chain antibody sequence and/or the full length light chain antibody sequence to create at least one altered antibody sequence; and expressing the altered antibody sequence as a protein.
  • the altered antibody sequence can also be prepared by screening antibody libraries having fixed CDR3 sequences selected among the group consisting of SEQ ID NO: 29-42 and SEQ ID NO: 71-84 or minimal essential binding determinants as described in US2005/0255552 and diversity on CDRl and CDR2 sequences.
  • the screening can be performed according to any screening technology appropriate for screening antibodies from antibody libraries, such as phage display technology.
  • Standard molecular biology techniques can be used to prepare and express the altered antibody sequence.
  • the antibody encoded by the altered antibody sequence(s) is one that retains one, some or all of the functional properties of the anti-ActRIIB antibodies described herein, which functional properties include, but are not limited to, specifically binding to human ActRIIB and inhibition of Smad activation.
  • the altered antibody may exhibit one or more, two or more, or three or more of the functional properties discussed above.
  • the functional properties of the altered antibodies can be assessed using standard assays available in the art and/or described herein, such as those set forth in the Examples (e.g. ELISAs).
  • mutations can be introduced randomly or selectively along all or part of an anti-ActRIIB antibody coding sequence and the resulting modified anti-ActRIIB antibodies can be screened for binding activity and/or other functional properties as described herein.
  • Mutational methods have been described in the art.
  • WO02/092780 describes methods for creating and screening antibody mutations using saturation mutagenesis, synthetic ligation assembly, or a combination thereof.
  • WO03/074679 describes methods of using computational screening methods to optimize physiochemical properties of antibodies.
  • Another aspect of the invention pertains to nucleic acid molecules that encode the antibodies of the invention.
  • Examples of full length light chain nucleotide sequences optimized for expression in a mammalian cell are shown in SEQ ID NOs: 161-165 and 171-175.
  • Examples of full length heavy chain nucleotide sequences optimized for expression in a mammalian cell are shown in SEQ ID NOs: 166-170 and 176-180.
  • the nucleic acids may be present in whole cells, in a cell lysate, or may be nucleic acids in a partially purified or substantially pure form.
  • a nucleic acid is "isolated” or “rendered substantially pure” when purified away from other cellular components or other contaminants, e.g. other cellular nucleic acids or proteins, by standard techniques, including alkaline/SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis and others well known in the art. See, F. Ausubel, et al., ed. 1987 Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York.
  • a nucleic acid of the invention can be, for example, DNA or RNA and may or may not contain intronic sequences.
  • the nucleic acid is a cDNA molecule.
  • the nucleic acid may be present in a vector such as a phage display vector, or in a recombinant plasmid vector.
  • the invention also provides the vectors referred to as pBW522 and pBW524 (deposited at DSMZ, Inhoffenstr. 7B, D-38124 Braunschweig, Germany on 18 August 2009 under deposit numbers DSM22873 and DSM22874, respectively).
  • Nucleic acids of the invention can be obtained using standard molecular biology techniques.
  • hybridomas e.g. hybridomas prepared from transgenic mice carrying human immunoglobulin genes as described further below
  • cDNAs encoding the light and heavy chains of the antibody made by the hybridoma can be obtained by standard PCR amplification or cDNA cloning techniques.
  • nucleic acid encoding the antibody can be recovered from various phage clones that are members of the library. Also included within the scope of the invention are variant nucleic acid sequences that comprise one or more deletions, additions or substitutions.
  • the invention comprises one or more of SEQ ID NOs: 113-140 or 161-180, which comprises a conservative nucleotide substitution. Due to the degeneracy of the genetic code, an amino acid may be encoded by more than one codon. Thus, it is possible to amend the nucleotide sequence, while the translated amino acid sequence remains the same.
  • V H and V L segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example to convert the variable region genes to full-length antibody chain genes, to Fab fragment genes or to an scFv gene.
  • a V L - or V ⁇ -encoding DNA fragment is operatively linked to another DNA molecule, or to a fragment encoding another protein, such as an antibody constant region or a flexible linker.
  • the term "operatively linked”, as used in this context, is intended to mean that the two DNA fragments are joined in a functional manner, for example, such that the amino acid sequences encoded by the two DNA fragments remain in-frame, or such that the protein is expressed under control of a desired promoter.
  • the isolated DNA encoding the V H region can be converted to a full-length heavy chain gene by operatively linking the V ⁇ -encoding DNA to another DNA molecule encoding heavy chain constant regions (CHl, CH2 and CH3).
  • heavy chain constant regions CH2 and CH3
  • the sequences of human heavy chain constant region genes are known in the art (see e.g. Kabat, E. A., et al. [supra]) and DNA fragments encompassing these regions can be obtained by standard PCR amplification.
  • the heavy chain constant region can be an IgGl, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region.
  • the heavy chain contstant region is selected among IgGl isotypes.
  • the V ⁇ -encoding DNA can be operatively linked to another DNA molecule encoding only the heavy chain CHl constant region.
  • the isolated DNA encoding the V L region can be converted to a full-length light chain gene (as well as to a Fab light chain gene) by operatively linking the V L -encoding DNA to another DNA molecule encoding the light chain constant region, CL.
  • the sequences of human light chain constant region genes are known in the art (see e.g. Kabat, E. A., et al. [supra]) and DNA fragments encompassing these regions can be obtained by standard PCR amplification.
  • the light chain constant region can be a kappa or a lambda constant region.
  • the V H - and V L -encoding DNA fragments are operatively linked to another fragment encoding a flexible linker, e.g. encoding the amino acid sequence (Gly4 -Ser) 3 , such that the V H and V L sequences can be expressed as a contiguous single-chain protein, with the V L and V H regions joined by the flexible linker (see e.g. Bird et al., 1988 Science 242:423- 426; Huston et al., 1988 Proc. Natl. Acad. Sci. USA 85:5879-5883; McCafferty et al., 1990 Nature 348:552-554).
  • a flexible linker e.g. encoding the amino acid sequence (Gly4 -Ser) 3
  • nucleic acids of the invention may be used in gene delivery. That is, nucleic acid encoding the polypeptides (antibodies or functional proteins) of the invention may be directly delivered to a patient for translation in the patient.
  • the nucleic acid is typically "packaged" for administration to a patient.
  • Gene delivery vehicles may be non-viral, such as liposomes, or replication-deficient viruses, such as adenovirus as described by Berkner, K.L., in Curr. Top. Microbiol. Immunol., 158, 39-66 (1992) or adeno- associated virus (AAV) vectors as described by Muzyczka, N., in Curr. Top. Microbiol. Immunol., 158, 97-129 (1992) and U.S. Patent No. 5,252,479.
  • a retrovirus such as a lentivirus may be used.
  • a nucleic acid molecule encoding a polypeptide of the invention may be engineered for expression in a replication-defective retroviral vector.
  • This expression construct may then be isolated and introduced into a packaging cell transduced with a retroviral plasmid vector containing RNA encoding the polypeptide, such that the packaging cell now produces infectious viral particles containing the gene of interest.
  • These producer cells may be administered to a subject for engineering cells in vivo and expression of the polypeptide in vivo (see Chapter 20, Gene Therapy and other Molecular Genetic-based Therapeutic Approaches, (and references cited therein) in Human Molecular Genetics (1996), T Strachan and A P Read, BIOS Scientific Publishers Ltd).
  • Another approach is the administration of "naked DNA" in which the therapeutic gene is directly injected into the bloodstream or muscle tissue.
  • Monoclonal antibodies can be produced by a variety of techniques, including conventional monoclonal antibody methodology e.g. the standard somatic cell hybridization technique of Kohler and Milstein (1975 Nature 256: 495). Many techniques for producing monoclonal antibody can be employed e.g. viral or oncogenic transformation of B lymphocytes.
  • An animal system for preparing hybridomas is the murine system.
  • Hybridoma production in the mouse is a well established procedure. Immunization protocols and techniques for isolation of immunized splenocytes for fusion are known in the art. Fusion partners (e.g. murine myeloma cells) and fusion procedures are also known.
  • Chimeric or humanized antibodies of the present invention can be prepared based on the sequence of a murine monoclonal antibody prepared as described above. DNA encoding the heavy and light chain immunoglobulins can be obtained from the murine hybridoma of interest and engineered to contain non-murine (e.g. human) immunoglobulin sequences using standard molecular biology techniques.
  • the murine variable regions can be linked to human constant regions using methods known in the art (see e.g. US4,816,567).
  • the murine CDR regions can be inserted into a human framework using methods known in the art (see e.g. U.S. Patent No. 5225539; 5530101; 5585089; 5693762 and 6180370).
  • the antibodies of the invention are human monoclonal antibodies. Such human monoclonal antibodies directed against ActRIIB can be generated using transgenic or transchromosomic mice carrying parts of the human immune system rather than the mouse system.
  • HuMAb mice ® (Medarex, Inc.) contains human immunoglobulin gene miniloci that encode un-rearranged human heavy ( ⁇ and ⁇ ) and K light chain immunoglobulin sequences, together with targeted mutations that inactivate the endogenous ⁇ and K chain loci (see e.g. Lonberg, et al., 1994 Nature 368(6474): 856-859).
  • mice exhibit reduced expression of mouse IgM or K, and in response to immunization, the introduced human heavy and light chain transgenes undergo class switching and somatic mutation to generate high affinity human IgG ⁇ monoclonal (Lonberg, N. et al., 1994 [supra]; reviewed in Lonberg, N., 1994 Handbook of Experimental Pharmacology 113:49-101; Lonberg, N. and Huszar, D., 1995 Intern. Rev. Immunol.13: 65-93, and Harding, F. and Lonberg, N., 1995 Ann. N. Y. Acad. Sci. 764:536-546).
  • HuMAb mice and the genomic modifications carried by such mice, is further described in Taylor, L.
  • human antibodies of the invention can be raised using a mouse that carries human immunoglobulin sequences on transgenes and transchomosomes such as a mouse that carries a human heavy chain transgene and a human light chain transchromosome.
  • KM mice a mouse that carries a human heavy chain transgene and a human light chain transchromosome.
  • transgenic animal systems expressing human immunoglobulin genes are available in the art and can be used to raise anti-ActRIIB antibodies of the invention.
  • an alternative transgenic system referred to as the Xenomouse (Abgenix, Inc.) can be used.
  • Such mice are described in, e.g. U.S. Patent Nos. 5,939,598; 6,075,181; 6,114,598; 6, 150,584 and 6,162,963.
  • mice carrying both a human heavy chain transchromosome and a human light chain tranchromosome referred to as "TC mice” can be used; such mice are described in Tomizuka et al., 2000 Proc. Natl. Acad. Sci. USA 97:722-727.
  • cows carrying human heavy and light chain transchromosomes have been described in the art (Kuroiwa et al., 2002 Nature Biotechnology 20:889-894) and can be used to raise anti-ActRIIB antibodies of the invention.
  • Human recombinant antibodies of the invention can also be prepared using phage display methods for screening libraries of human immunoglobulin genes. Such phage display methods for isolating human antibodies are established in the art or described in the examples below. See for example: U.S. Patent Nos. 5,223,409; 5,403,484; 5,571,698; 5,427,908; 5,580,717; 5,969,108; 6,172,197; 5,885,793; 6,521,404; 6,544,731; 6,555,313; 6,582,915 and 6,593,081.
  • Human monoclonal antibodies of the invention can also be prepared using SCID mice into which human immune cells have been reconstituted such that a human antibody response can be generated upon immunization. Such mice are described in, for example, U.S. Patent Nos. 5,476,996 and 5,698,767.
  • hybridomas producing human monoclonal antibodies of the invention splenocytes and/or lymph node cells from immunized mice can be isolated and fused to an appropriate immortalized cell line, such as a mouse myeloma cell line.
  • an appropriate immortalized cell line such as a mouse myeloma cell line.
  • the resulting hybridomas can be screened for the production of antigen- specific antibodies.
  • single cell suspensions of splenic lymphocytes from immunized mice can be fused to one-sixth the number of P3X63- Ag8.653 nonsecreting mouse myeloma cells (ATCC, CRL 1580) with 50% PEG.
  • Cells are plated at approximately 2 x 145 in flat bottom microtiter plates, followed by a two week incubation in selective medium containing 20% fetal Clone Serum, 18% "653" conditioned media, 5% origen (IGEN), 4 mM L-glutamine, 1 mM sodium pyruvate, 5mM HEPES, 0:055 mM 2-mercaptoethanol, 50 units/ml penicillin, 50 mg/ml streptomycin, 50 mg/ml gentamycin and IX HAT (Sigma; the HAT is added 24 hours after the fusion). After approximately two weeks, cells can be cultured in medium in which the HAT is replaced with HT.
  • selective medium containing 20% fetal Clone Serum, 18% "653" conditioned media, 5% origen (IGEN), 4 mM L-glutamine, 1 mM sodium pyruvate, 5mM HEPES, 0:055 mM 2-mercaptoethanol, 50 units/m
  • selected hybridomas can be grown in two -liter spinner- flasks for monoclonal antibody purification.
  • Supernatants can be filtered and concentrated before affinity chromatography with protein A-sepharose (Pharmacia).
  • Eluted IgG can be checked by gel electrophoresis and high performance liquid chromatography to ensure purity.
  • the buffer solution can be exchanged into PBS, and the concentration can be determined by OD 28 o using 1.43 extinction coefficient.
  • the monoclonal antibodies can be aliquoted and stored at -8O 0 C.
  • Antibodies of the invention also can be produced in a host cell transfectoma using, for example, a combination of recombinant DNA techniques and gene transfection methods as is well known in the art (e.g. Morrison, S. (1985) Science 229:1202).
  • DNAs encoding partial or full-length light and heavy chains can be obtained by standard molecular biology techniques (e.g. PCR amplification or cDNA cloning using a hybridoma that expresses the antibody of interest) and the DNAs can be inserted into expression vectors such that the genes are operatively linked to transcriptional and translational control sequences.
  • operatively linked is intended to mean that an antibody gene is ligated into a vector such that transcriptional and translational control sequences within the vector serve their intended function of regulating the transcription and translation of the antibody gene.
  • the expression vector and expression control sequences are chosen to be compatible with the expression host cell used.
  • the antibody light chain gene and the antibody heavy chain gene can be inserted into separate vector or, more typically, both genes are inserted into the same expression vector.
  • the antibody genes are inserted into the expression vector by standard methods (e.g. ligation of complementary restriction sites on the antibody gene fragment and vector, or blunt end ligation if no restriction sites are present).
  • the light and heavy chain variable regions of the antibodies described herein can be used to create full-length antibody genes of any antibody isotype by inserting them into expression vectors already encoding heavy chain constant and light chain constant regions of the desired isotype such that the V H segment is operatively linked to the CH segment(s) within the vector and the V L segment is operatively linked to the CL segment within the vector.
  • the recombinant expression vector can encode a signal peptide that facilitates secretion of the antibody chain from a host cell.
  • the antibody chain gene can be cloned into the vector such that the signal peptide is linked in frame to the amino terminus of the antibody chain gene.
  • the signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e. a signal peptide from a non-immunoglobulin protein).
  • the recombinant expression vectors of the invention carry regulatory sequences that control the expression of the antibody chain genes in a host cell.
  • the term "regulatory sequence” is intended to include promoters, enhancers and other expression control elements (e.g. polyadenylation signals) that control the transcription or translation of the antibody chain genes.
  • Such regulatory sequences are described, for example, in Goeddel (Gene Expression Technology. Methods in Enzymology 185, Academic Press, San Diego, CA 1990). It will be appreciated by those skilled in the art that the design of the expression vector, including the selection of regulatory sequences, may depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc.
  • Regulatory sequences for mammalian host cell expression include viral elements that direct high levels of protein expression in mammalian cells, such as promoters and/or enhancers derived from cytomegalovirus (CMV), Simian Virus 40 (SV40), adenovirus (e.g. the adenovirus major late promoter (AdMLP)), and polyoma.
  • CMV cytomegalovirus
  • SV40 Simian Virus 40
  • AdMLP adenovirus major late promoter
  • polyoma e.g. the adenovirus major late promoter (AdMLP)
  • nonviral regulatory sequences may be used, such as the ubiquitin promoter or P-globin promoter.
  • regulatory elements composed of sequences from different sources such as the SRa promoter system, which contains sequences from the SV40 early promoter and the long terminal repeat of human T cell leukemia virus type 1 (Takebe, Y.
  • the recombinant expression vectors of the invention may carry additional sequences, such as sequences that regulate replication of the vector in host cells (e.g. origins of replication) and selectable marker genes.
  • the selectable marker gene facilitates selection of host cells into which the vector has been introduced (see, e.g. U.S. Patent Nos. 4,399,216, 4,634,665 and 5,179,017).
  • the selectable marker gene confers resistance to drugs, such as G418, hygromycin or methotrexate, on a host cell into which the vector has been introduced.
  • Selectable marker genes include the dihydrofolate reductase (DHFR) gene (for use in dhfr- host cells with methotrexate selection/amplification) and the neo gene (for G418 selection).
  • DHFR dihydrofolate reductase
  • neo gene for G418 selection.
  • the expression vector(s) encoding the heavy and light chains is transfected into a host cell by standard techniques.
  • the various forms of the term "transfection" are intended to encompass a wide variety of techniques commonly used for the introduction of exogenous DNA into a prokaryotic or eukaryotic host cell, e.g. electroporation, calcium-phosphate precipitation, DEAE-dextran transfection and the like.
  • Mammalian host cells for expressing the recombinant antibodies of the invention include Chinese Hamster Ovary (CHO cells) (including dhfr- CHO cells, described Urlaub and Chasin, 1980 Proc. Natl. Acad. Sci. USA 77:4216-4220 used with a DH FR selectable marker, e.g. as described in RJ. Kaufman and P.A. Sharp, 1982 MoI. Biol. 159:601-621), NSO myeloma cells, COS cells and SP2 cells.
  • the host cells are CHO KlPD cells.
  • mammalian host cells for expressing the recombinant antibodies of the invention include mammalian cell lines deficient for FUT8 gene expression, for example as described in US6,946,292B2.
  • the antibodies are produced by culturing the host cells for a period of time sufficient to allow for expression of the antibody in the host cells or secretion of the antibody into the culture medium in which the host cells are grown. Antibodies can be recovered from the culture medium using standard protein purification methods.
  • the present invention features an anti-ActRIIB antibody, or a fragment thereof, conjugated to a therapeutic moiety, such as a cytotoxin, a drug (e.g. an immunosuppressant) or a radiotoxin.
  • a therapeutic moiety such as a cytotoxin, a drug (e.g. an immunosuppressant) or a radiotoxin.
  • cytotoxin e.g. an immunosuppressant
  • radiotoxin e.g. an immunosuppressant
  • immunotoxins e.g. an immunosuppressant
  • a cytotoxin or cytotoxic agent includes any agent that is detrimental to (e.g. kills) cells.
  • Cytotoxins can be conjugated to antibodies of the invention using linker technology available in the art.
  • linker types that have been used to conjugate a cytotoxin to an antibody include, but are not limited to, hydrazones, thioethers, esters, disulfides and peptide-containing linkers.
  • a linker can be chosen that is, for example, susceptible to cleavage by low pH within the lysosomal compartment or susceptible to cleavage by proteases, such as proteases preferentially expressed in tumor tissue such as cathepsins (e.g. cathepsins B, C, D).
  • cathepsins e.g. cathepsins B, C, D
  • Antibodies of the present invention also can be conjugated to a radioactive isotope to generate cytotoxic radiopharmaceuticals, also referred to as radioimmunoconjugates.
  • radioactive isotopes that can be conjugated to antibodies for use diagnostically or therapeutically include, but are not limited to, iodine 131 , indium 111 , yttrium 90 , and lutetium 177 .
  • Methods for preparing radioimmunconjugates are established in the art. Examples of radioimmunoconjugates are commercially available, including Zevalin (DEC Pharmaceuticals) and Bexxar (Corixa Pharmaceuticals), and similar methods can be used to prepare radioimmunoconjugates using the antibodies of the invention.
  • the antibody conjugates of the invention can be used to modify a given biological response, and the drug moiety is not to be construed as limited to classical chemical therapeutic agents.
  • the drug moiety may be a protein or polypeptide possessing a desired biological activity.
  • proteins may include, for example, an enzymatically active toxin, or active fragment thereof, such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor or interferon- ⁇ ; or, biological response modifiers such as, for example, lymphokines, interleukin-1 ("IL-I”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors.
  • IL-I interleukin-1
  • IL-2 interleukin-2
  • IL-6 interleukin-6
  • the present invention features bispecific or multispecific molecules comprising an anti-ActRIIB antibody, or a fragment thereof, of the invention.
  • An antibody of the invention, or antigen-binding regions thereof can be derivatized or linked to another functional molecule, e.g. another peptide or protein (e.g. another antibody or ligand for a receptor) to generate a bispecific molecule that binds to at least two different binding sites or target molecules.
  • the antibody of the invention may in fact be derivatized or linked to more than one other functional molecule to generate multi- specific molecules that bind to more than two different binding sites and/or target molecules; such multi-specific molecules are also intended to be encompassed by the term "bispecific molecule" as used herein.
  • an antibody of the invention can be functionally linked (e.g. by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other binding molecules, such as another antibody, antibody fragment, peptide or binding mimetic, such that a bispecific molecule results.
  • the present invention includes bispecific molecules comprising at least one first binding specificity for ActRIIB and a second binding specificity for a second target epitope.
  • the second target epitope may be another epitope of ActRIIB different from the first target epitope.
  • the molecule can further include a third binding specificity, in addition to the first and second target epitope.
  • the bispecific molecules of the invention comprise as a binding specificity at least one antibody, or an antibody fragment thereof, including, e.g. an Fab, Fab', F(ab') 2 , Fv, or a single chain Fv.
  • the antibody may also be a light chain or heavy chain dimer, or any minimal fragment thereof such as a Fv or a single chain construct as described in Ladner et al. US4,946,778, the contents of which is expressly incorporated by reference.
  • Other antibodies which can be employed in the bispecific molecules of the invention are murine, chimeric and humanized monoclonal antibodies.
  • the bispecific molecules of the present invention can be prepared by conjugating the constituent binding specificities, using methods known in the art. For example, each binding specificity of the bispecific molecule can be generated separately and then conjugated to one another. When the binding specificities are proteins or peptides, a variety of coupling or cross-linking agents can be used for covalent conjugation.
  • cross-linking agents examples include protein A, carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SATA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohaxane-1-carboxylate (sulfo- SMCC) (see e.g. Karpovsky et al., 1984 J. Exp. Med.
  • Conjugating agents are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, IL).
  • binding specificities are antibodies, they can be conjugated by sulfhydryl bonding of the C-terminus hinge regions of the two heavy chains.
  • the hinge region is modified to contain an odd number of sulfhydryl residues, for example one, prior to conjugation.
  • both binding specificities can be encoded in the same vector and expressed and assembled in the same host cell.
  • This method is particularly useful where the bispecific molecule is a mAb x mAb, mAb x Fab, Fab x F(ab') 2 or ligand x Fab fusion protein.
  • a bispecific molecule of the invention can be a single chain molecule comprising one single chain antibody and a binding determinant, or a single chain bispecific molecule comprising two binding determinants. Bispecific molecules may comprise at least two single chain molecules. Methods for preparing bispecific molecules are described for example in U.S.
  • a labeled reagent e.g. an antibody
  • the present invention provides multivalent antibodies comprising at least two identical or different antigen-binding portions of the antibodies of the invention binding to ActRIIB.
  • the multivalent antibodies provides at least two, three or four antigen -binding portions of the antibodies.
  • the antigen-binding portions can be linked together via protein fusion or covalent or non covalent linkage. Alternatively, methods of linkage have been described for the bispecific molecules. Tetravalent compounds can be obtained for example by cross-linking antibodies of the antibodies of the invention with an antibody that binds to the constant regions of the antibodies of the invention, for example the Fc or hinge region.
  • compositions in another aspect, provides a composition, e.g. a pharmaceutical composition, containing one or a combination of monoclonal antibodies, or antigen-binding portion(s) thereof, of the present invention, formulated together with a pharmaceutically acceptable carrier.
  • a pharmaceutical composition of the invention can comprise a combination of antibodies that bind to different epitopes on the target antigen or that have complementary activities.
  • compositions of the invention also can be administered in combination therapy, i.e. combined with other agents.
  • the combination therapy can include an anti- ActRIIB antibody of the present invention combined with at least one other muscle mass/strength increasing agent, for example, IGF-I, IGF-2 or variants of IGF-I or IGF-2, an anti-myostatin antibody, a myostatin propeptide, a myostatin decoy protein that binds ActRIIB but does not activate it, a beta 2 agonist, a Ghrelin agonist, a SARM, GH agonists/mimetics or follistatin.
  • therapeutic agents that can be used in combination therapy are described in greater detail below in the section on uses of the antibodies of the invention.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.
  • the carrier should be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g. by injection or infusion).
  • the active compound i.e. antibody, immunoconjuage, or bispecific molecule, may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound.
  • the pharmaceutical compounds of the invention may include one or more pharmaceutically acceptable salts.
  • a "pharmaceutically acceptable salt” refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects (see e.g. Berge, S.M., et al., 1977 J. Pharm. Sci. 66:1-19). Examples of such salts include acid addition salts and base addition salts.
  • Acid addition salts include those derived from nontoxic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous and the like, as well as from nontoxic organic acids such as aliphatic mono- and di-carboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like.
  • nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous and the like
  • nontoxic organic acids such as aliphatic mono- and di-carboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like.
  • Base addition salts include those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium and the like, as well as from nontoxic organic amines, such as N,N'-dibenzylethylenediamine, N- methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like.
  • a pharmaceutical composition of the invention also may include a pharmaceutically acceptable anti-oxidant.
  • pharmaceutically acceptable antioxidants include: water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.
  • water soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like
  • oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole
  • aqueous and nonaqueous carriers examples include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate.
  • polyols such as glycerol, propylene glycol, polyethylene glycol, and the like
  • vegetable oils such as olive oil
  • injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of presence of microorganisms may be ensured both by sterilization procedures, supra, and by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as, aluminum monostearate and gelatin.
  • Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • the use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the invention is contemplated. Supplementary active compounds can also be incorporated into the compositions.
  • compositions typically must be sterile and stable under the conditions of manufacture and storage.
  • the composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption for example, monostearate salts and gelatin.
  • Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of agents enumerated above, as required, followed by sterilization microfiltration.
  • dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other agents from those enumerated above.
  • the methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active agent plus any additional desired agent from a previously sterile-filtered solution thereof.
  • the amount of active agent which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, and the particular mode of administration.
  • the amount of active agent which can be combined with a carrier material to produce a single dosage form will generally be that amount of the composition which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.01 per cent to about ninety-nine percent of active agent, from about 0.1 per cent to about 70 per cent, or from about 1 percent to about 30 percent of active agent in combination with a pharmaceutically acceptable carrier.
  • Dosage regimens are adjusted to provide the optimum desired response (e.g. a therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage.
  • Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.
  • the dosage ranges from about 0.0001 to 100 mg/kg, and more usually 0.01 to 5 mg/kg, of the host body weight.
  • dosages can be 0.3 mg/kg body weight, 1 mg/kg body weight, 3 mg/kg body weight, 5 mg/kg body weight or 10 mg/kg body weight or within the ranges of 1-10 mg/kg or 3-7 mg/kg.
  • An exemplary treatment regime entails administration once per week, once every two weeks, once every three weeks, once every four weeks, once a month, once every 3 months or once every three to 6 months.
  • the antibody may be administered about once a year or once only. Such administration may be carried out intraveneously or subcutaneously.
  • Dosage regimens for an anti-ActRIIB antibody of the invention include 1 mg/kg body weight or 3 mg/kg body weight by intravenous administration, with the antibody being given using one of the following dosing schedules: every four weeks for six dosages, then every three months; every three weeks; 3 mg/kg body weight once followed by 1 mg/kg body weight every three weeks.
  • the dosage should be one that causes an upregulation of muscle mass and/or strength.
  • the effect is on skeletal muscle.
  • the dosage causes muscle hypertrophy with no more than a proportional increase in the size of internal organs (e.g. heart, lungs, liver, kidneys). Such a proportional increase may be compared by measuring either mass or volume.
  • two or more monoclonal antibodies with different binding specificities are administered simultaneously, in which case the dosage of each antibody administered falls within the ranges indicated.
  • Antibody is usually administered on multiple occasions. Intervals between single dosages can be, for example, weekly, monthly, every three months, every six months or yearly. Intervals can also be irregular as indicated by measuring blood levels of antibody to the target antigen in the patient. In some methods, dosage is adjusted to achieve a plasma antibody concentration of about 1-1000 ⁇ g/ml and in some methods about 25-300 ⁇ g/ml.
  • an ActRIIB antibody of the invention could be co-administered with an anti- myostatin antibody.
  • antibody can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the patient. In general, human antibodies show the longest half-life, followed by humanized antibodies, chimeric antibodies, and nonhuman antibodies. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated or until the patient shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime.
  • Actual dosage levels of the active agents in the pharmaceutical compositions of the present invention may be varied so as to obtain an amount of the active agent which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
  • the selected dosage level will depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.
  • a "therapeutically effective dosage" of an anti-ActRIIB antibody of the invention can result in a decrease in severity of disease symptoms, an increase in frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction i.e. an increase in muscle mass and/or strength.
  • a composition of the present invention can be administered by one or more routes of administration using one or more of a variety of methods known in the art. As will be appreciated by the skilled artisan, the route and/or mode of administration will vary depending upon the desired results. Routes of administration for antibodies of the invention include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion.
  • parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal injection and infusion.
  • the antibody is administered intravenously. In another embodiment the antibody is administered subcutaneously.
  • an antibody of the invention can be administered by a nonparenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically.
  • a nonparenteral route such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically.
  • the active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems.
  • a controlled release formulation including implants, transdermal patches, and microencapsulated delivery systems.
  • Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, e.g. Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.
  • Therapeutic compositions can be administered with medical devices known in the art.
  • a therapeutic composition of the invention can be administered with a needleless hypodermic injection device, such as the devices shown in U.S. Patent Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556.
  • a needleless hypodermic injection device such as the devices shown in U.S. Patent Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556.
  • Examples of well known implants and modules useful in the present invention include: U.S. Patent No. 4,487,603, which shows an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Patent No. 4,486,194, which shows a therapeutic device for administering medicants through the skin; U.S. Patent No.
  • the human monoclonal antibodies of the invention can be formulated to ensure proper distribution in vivo.
  • the blood-brain barrier excludes many highly hydrophilic compounds.
  • the therapeutic compounds of the invention cross the BBB (if desired)
  • they can be formulated, for example, in liposomes.
  • liposomes For methods of manufacturing liposomes, see, e.g. U.S. Patents 4,522,811; 5,374,548; and 5,399,331.
  • the liposomes may comprise one or more moieties which are selectively transported into specific cells or organs, thus enhance targeted drug delivery (see, e.g. V.V. Ranade, 1989 J. Clin Pharmacol. 29:685).
  • Exemplary targeting moieties include folate or biotin (see, e.g. U.S. Patent 5,416,016); mannosides (Umezawa et al., 1988 Biochem. Biophys. Res. Commun. 153:1038); antibodies (P.G. Bloeman et al., 1995 FEBS Lett. 357:140; M. Owais et al., 1995 Antimicrob. Agents Chemother. 39:180); surfactant protein A receptor (Briscoe et al., 1995 Am. J. Physiol.1233: 134); pl20 (Schreier et al., 1994 J. Biol. Chem. 269:9090); see also K. Keinanen; M.L. Laukkanen, 1994 FEBSLett. 346:123; JJ. Killion; LJ. Fidler, 1994 Imrnunomethods 4:273.
  • biotin see, e.g. U.S. Patent 5,416,
  • the antibodies of the present invention have in vitro and in vivo diagnostic and therapeutic utilities.
  • these molecules can be administered to cells in culture, e.g. in vitro or ex vivo, or in a subject, e.g. in vivo, to treat, prevent or diagnose a variety of disorders.
  • the antibodies may be used in both the treatment of disease, prophylaxis and for delaying the onset of disease symptoms.
  • the term "subject" as used herein is intended to include human and non- human animals.
  • Non-human animals include all vertebrates, e.g. mammals and non-mammals, such as non-human primates, sheep, dogs, cats, cows, horses, chickens, amphibians, and reptiles.
  • the invention provides a method of treating a patient suffering from a pathological disorder (such as a muscle wasting disease or disorder) comprising administering a therapeutically effective amount of an anti-ActRIIB antibody.
  • the invention also provides an anti-ActRIIB antibody for use in therapy.
  • the invention also provides use of an anti-ActRIIB antibody in the manufacture of a medicament for the treatment of a pathological disorder.
  • the methods are particularly suitable for treating, preventing, ameliorating or diagnosing pathological disorders.
  • a "pathological disorder” includes, but is not limited to, musculoskeletal diseases or disorders, such as muscle atrophy.
  • muscle atrophy There are many causes of muscle atrophy, including as a result of treatment with a glucocorticoid such as Cortisol, dexamethasone, betamethasone, prednisone, methylprednisolone, or prednisolone.
  • the muscle atrophy can also be a result of denervation due to nerve trauma or a result of degenerative, metabolic, or inflammatory neuropathy (e.g., Guillian-Barre syndrome, peripheral neuropathy, or exposure to environmental toxins or drugs).
  • the muscle atrophy can be a result of myopathy, such as myotonia; a congential myopathy, including nemalene myopathy, multi/minicore myopathy and myotubular (centronuclear) myopathy; mitochondrial myopathy; familial periodic paralysis; inflammatory myopathy; metabolic myopathy, such as caused by a glycogen or lipid storage disease; dermatomyositisis; polymyositis; inclusion body myositis; myositis ossificans; rhabdomyolysis and myoglobinurias.
  • myopathy such as myotonia; a congential myopathy, including nemalene myopathy, multi/minicore myopathy and myotubular (centronuclear) myopathy; mitochondrial myopathy; familial periodic paralysis; inflammatory myopathy; metabolic myopathy, such as caused by a glycogen or lipid storage disease; dermatomyositisis; polymyositis; inclusion body myositis; my
  • the myopathy may be caused by a muscular dystrophy syndrome, such as Duchenne, Becker, myotonic, fascioscapulohumeral, Emery-Dreifuss, oculopharyngeal, scapulohumeral, limb girdle, Fukuyama, a congenital muscular dystrophy, or hereditary distal myopathy.
  • the musculoskeletal disease can also be osteoporosis, a bone fracture, short stature, or dwarfism.
  • the muscle atrophy can be a result of an adult motor neuron disease, infantile spinal muscular atrophy, amyotrophic lateral sclerosis, juvenile spinal muscular atrophy, autoimmune motor neuropathy with multifocal conductor block, paralysis due to stroke or spinal cord injury, skeletal immobilization due to trauma, prolonged bed rest, voluntary inactivity, involuntary inactivity, metabolic stress or nutritional insufficiency, cancer, AIDS, fasting, a thyroid gland disorder, diabetes, benign congenital hypotonia, central core disease, burn injury, chronic obstructive pulmonary disease, liver diseases (examples such as fibrosis, cirrhosis), sepsis, renal failure, congestive heart failure, ageing, space travel or time spent in a zero gravity environment.
  • adult motor neuron disease infantile spinal muscular atrophy, amyotrophic lateral sclerosis, juvenile spinal muscular atrophy, autoimmune motor neuropathy with multifocal conductor block, paralysis due to stroke or spinal cord injury, skeletal immobilization due to trauma, prolonged
  • age-related conditions examples include, sarcopenia, skin atrophy, muscle wasting, brain atrophy, atherosclerosis, arteriosclerosis, pulmonary emphysema, osteoporosis, osteoarthritis, immunologic incompetence, high blood pressure, dementia, Huntington's disease, Alzheimer's disease, cataracts, age-related macular degeneration, prostate cancer, stroke, diminished life expectancy, frailty, memory loss, wrinkles, impaired kidney function, and age- related hearing loss; metabolic disorders, including Type II Diabetes, Metabolic Syndrome, hyperglycemia, and obesity.
  • patients may simultaneously suffer from one or more of these conditions, for example, sarcopenia and pulmonary emphysema, or sarcopenia and impaired kidney function.
  • pathological disorders include acute and/or chronic renal disease or failure, liver fibrosis or cirrhosis, cancer such as breast cancer, Parkinson's Disease; conditions associated with neuronal death, such as ALS, brain atrophy, or dementia and anemia.
  • Further conditions include cachexia, cachexia associated with a rheumatoid arthritis and cachexia associated with cancer.
  • the "pathological disorders" recited herein include liver, kidney and pulmonary fibrosis and cancers examplified by but not restricted to rhabdomyosarcomas, bone-loss inducing cancers, hepatocellular carcinomas, gastrointestinal cancers.
  • the prevention may be complete, e.g., the total absence of an age-related condition or metabolic disorder.
  • the prevention may also be partial, such that the likelihood of the occurrence of the age-related condition or metabolic disorder in a subject is less likely to occur than had the subject not received an antibody of the present invention.
  • An age-related condition as referred to herein may begin at the age of 50 years or older (i.e. 60, 70, 80 or older).
  • a patient may be pre-treated with an anti-ActRIIB antibody prior to an anticipated period of enforced rest/inactivity. Such a period may occur when a patient is admitted to hospital, for example for surgery to the hip or leg.
  • the inactivity may be localised, such as by casting of a broken limb or joint, or by administration of a paralytic agent.
  • the patient being treated has a fracture to a limb (i.e. leg or arm) or joint (i.e. knee or hip).
  • the patient being treated has a fracture to one or more of the humerus, radius, ulnar, a carpal, a metacarpal, the clavical, scapular, femur, os coxae, patella, tibia, fibula, talus, calcaneus, a tarsal, a metatarsal, the ischium or the ileum.
  • the patient being treated has undergone, or will undergo surgery on one or more of the following joints: knee, hip, ankle, shoulder, elbow. Such surgery includes hip replacement and knee replacement.
  • Atrophy due to immobilisation may occur quickly, but normally occurs slowly. Therefore, in one embodiment, the patient, joint or limb has been immobilised, or will be immobilised, for 2 weeks or longer (i.e. 3 weeks, 4 weeks, 6 weeks, 8 weeks or longer). In one embodiment, the patient, joint or limb has been immobilised, or will be immobilised, for 1-8 weeks, 2-6 weeks or 3-5 weeks.
  • the patient may be one who has not responded to previous bone anabolic treatments.
  • the patient may not have responded to treatment with IGF-I, IGF-2 or variants of IGF-I or IGF-2, an anti-myostatin antibody, a myostatin propeptide, a myostatin decoy protein that binds ActRIIB but does not activate it, a beta 2 agonist, a Ghrelin agonist, a SARM, GH agonists/mimetics or follistatin.
  • a simple way of measuring a patient's response to treatment may be timing how long it takes for a patient to climb a known height of stairs and comparing the results both before and after treatment.
  • the antibodies of the invention may be administered as the sole active agent or in conjunction with, e.g. as an adjuvant to or in combination to, other drugs e.g. IGF-I, IGF-2 or variants of IGF-I or IGF-2, an anti-myostatin antibody, a myostatin propeptide, a myostatin decoy protein that binds ActRIIB but does not activate it, a beta 2 agonist, a Ghrelin agonist, a SARM, GH agonists/mimetics or follistatin.
  • the antibodies of the invention may be used in combination with an IGF-I mimetic as disclosed in WO2007/146689.
  • the present invention provides in a yet further aspect:
  • a method as defined above comprising co-administration, e.g. concomitantly or in sequence, of a therapeutically effective amount of an ActRIIB antagonist, e.g. an antibody of the invention, and at least one second drug substance, said second drug substance being IGF-I, IGF-2 or variants of IGF-I or IGF-2, an anti-myostatin antibody, a myostatin propeptide, a myostatin decoy protein that binds ActRIIB but does not activate it, a beta 2 agonist, a Ghrelin agonist, a SARM, GH agonists/mimetics or follistatin.
  • the invention further provides a therapeutic combination, e.g.
  • kits comprising of a therapeutically effective amount of a) an ActRIIB antagonist, e.g. an antibody of the invention, and b) at least one second substance selected from an IGF-I, IGF-2 or variants of IGF-I or IGF- 2, an anti-myostatin antibody, a myostatin propeptide, a myostatin decoy protein that binds ActRIIB but does not activate it, a beta 2 agonist, a Ghrelin agonist, a SARM, GH agonists/mimetics or follistatin, e.g. as indicated above.
  • the kit may further comprise instructions for its administration.
  • dosages of the co-administered combination compound will of course vary depending on the type of co-drug employed, on the specific drug employed, on the condition being treated and so forth.
  • the antibodies of the invention are administered only to a patient population which is selected among patients suffering from muscle atrophy. In another embodiment, the antibodies of the invention are administered to patient populations suffering from skeletal muscle atrophy. In another embodiment, the antibodies of the invention are administered only to a patient population which is selected among a group of patients which respond to anti-ActRIIB treatment. Biomarkers that identify patients that have an increased likelihood of responding to anti-ActRIIB treatment may be any of the following without being limited to these: high levels of serum myostatin, GDF-Il or activins compared to a control patient. In one embodiment, the antibodies of the invention can be used to detect levels of ActRIIB, or levels of cells that contain ActRIIB.
  • the invention further provides methods for detecting the presence of ActRIIB (e.g.
  • kits consisting of the compositions (e.g. antibodies, human antibodies and bispecific molecules) of the invention and instructions for use.
  • the kit can further contain at least one additional reagent, or one or more additional antibodies of the invention (e.g. an antibody having a complementary activity which binds to an epitope on the target antigen distinct from the first antibody).
  • Kits typically include a label indicating the intended use of the contents of the kit.
  • the term label includes any writing, or recorded material supplied on or with the kit, or which otherwise accompanies the kit.
  • the kit may further comprise tools for diagnosing whether a patient belongs to a group that will response to an anti- ActRIIB antibody treatment, as defined above.
  • Such kits may comprise an antibody of the invention in lyophilised form, a diluent and instructions for use.
  • composition comprising X may consist exclusively of X or may include something additional e.g. X + Y.
  • FIGURES Figure 1 shows EC50 determination of MOR07079 by FACS titration on parental and ActRIIB transfected HEK293T/17 cell lines.
  • Figure 2 shows inhibition of myostatin-induced luciferase expression in a reporter gene assay by multiples of anti-ActRIIB Fabs at 2, 10 and 50 ⁇ g/ml.
  • Figure 3 shows IC50 determination of Fabs in myostatin-induced luciferase reporter gene assay.
  • Figure 4 shows antibody binding to primary human skeletal muscle cells.
  • Figure 5 shows IC50 determination of IgG in myostatin-induced inhibition of skeletal muscle differentiation assay.
  • Figure 6 shows mouse study: in vivo efficacy study in naive animals - 6 weeks treatment with MOR08159 or MOR08213 at 10mg/kg increased body and muscle weight. Changes are shown for (A) body weight, (B) Tibialis, (C) Gastrocnemius with plantaris, (D) Quadriceps and (E) Pectoralis.
  • Figure 7 shows mouse study: dose response in vivo efficacy study in naive animals - 6 weeks treatment with MOR08213 at 25, 5, 1 mg/kg, dose-dependently increases body and muscle weight. Changes are shown for (A) body weight, (B) Tibialis, (C) Gastrocnemius with plantaris, (D) Quadriceps and (E) Pectoralis.
  • Figure 8 shows a FACS output demonstrating cross-blocking between MOR08159 in the presence of MOR08213 (bold dashed) and MOR08159 alone (bold black), compared to isotype control alone (black) or isotype control in the presence of MOR08213 (dashed).
  • Figure 9 shows an overview of ActRIIB residues (SEQ ID NO: 181) to which MOR08159 binds, using various epitope determination techniques.
  • HEK293T/17 cells were maintained in DMEM containing 10% FBS, 2 mM L- glutamine, penicillin (50 IE/ml), and streptomycin (50 ⁇ g/ml). Cells were grown in an incubator at 37 0 C and 5% CO 2 and subcultured every 3-4 days. Cells were detached using AccutaseTM and then transferred into a new flask containing fresh medium.
  • HEK293T/17 cells stably transfected with CAGA- 12 luc were cultured as described above for parental HEK293T/17 cells but cell growth medium was supplemented with 4 mM L-glutamine and 3 ⁇ g/ml blasticidin in addition to FBS, penicillin and streptomycin.
  • Myostatin-Induced Luciferase Reporter Gene Assay To determine the capacity of anti-ActRIIB antibodies to inhibit myostatin-induced signaling, a reporter gene assay using the stable reporter cell line HEK293T/17 CAGA-12 luc was performed.
  • the CAGA-12 lucif erase reporter construct carries the lucif erase gene downstream of a minimal promoter and multiple CAGA boxes which are specific for phosphorylated Smad-2 and Smad-3. Addition of purified myostatin (but also of GDF-Il, activin or TGFB) induces Smad phosphorylation and thus binding to the CAGA-12 reporter and leads to luciferase gene expression.
  • HEK293T/17 CAGA-12 luc cells were detached as described and diluted in culture medium to a concentration of 2.5x10 5 cells/ml. Subsequently, 100 ⁇ l cells per well were seeded into flat-bottomed 96- well plates and incubated at 37 0 C and 5% CO 2 overnight.
  • the antibodies (Fab or IgG) and the recombinant human ActRIIB/Fc, which served as the positive control, were diluted in PBS to the desired concentrations. 20 ⁇ l of the antibody solutions were added to the seeded wells of the previous day and the cells cultivated for 1 hr to allow binding of the antibodies. Finally, 50 ng/ml myostatin was added to the wells and the cells further cultivated over night.
  • SPECIFICITYELISAs The specificity of anti-ActRIIB Fab antibodies to human ActRIIB and crossreactivity to human ActRIIA and mouse ActRIIB was evaluated in an ELISA setting. Additionally, binding to related receptors (counter-targets: human TGF- ⁇ RII/Fc (R&D systems), mouse TGF- ⁇ RI (ALK-5)/Fc (R&D systems), human Activin RIB (ALK-4)/Fc (R&D systems)) was determined. For this, 5 ⁇ g/ml (if not stated otherwise) of the recombinant proteins diluted in PBS were added to a black 96-well flat-bottom MaxiSorpTM plate and incubated over night at 4 0 C for coating.
  • hActRIIB/Fc-myostatin interaction ELISA was performed. For this, recombinant myostatin was diluted to 5 ⁇ g/ml in PBS and coated onto a black 96-well flat-bottom Maxisorp plate. The next morning the wells were blocked with MTBST. Meanwhile 50 ⁇ g/ml anti- ActRIIB Fabs were pre-incubated with 10 ⁇ g/ml ActRIIB/Fc in TBST for 1.5 hrs at room temperature and finally added to the coated and blocked wells (1.5 hr at room temperature).
  • Cells Stable human ActRIIA- and human ActRIIB-transfected HEK293T/17 cells generated using HEK293T/17 cells (ATCC) transfected with linearized pEGFP (Clontech) -ActRIIB (ECD) or -ActRIIA(ECD) and pPGK-puro (AddGene) using FuGENE ⁇ (Roche), were maintained in DMEM containing 10% FBS, 2 mM L-glutamine, penicillin (50 IE/ml), streptomycin (50 ⁇ g/ml) and puromycin (2 ⁇ g/ml). Cells were grown in an incubator at 37 0 C and 5% CO 2 and subcultured every 3-4 days. Cells were detached using AccutaseTM and then transferred into a new flask containing fresh media.
  • huSkMC Human skeletal muscle cells
  • GM growth medium
  • skBM skeletal muscle basal medium
  • FCS FCS
  • trypsin was neutralized with trypsin neutralizing solution.
  • Cells were centrifuged at 220 x g for 5 min and the pellet was resuspended in Skeletal Muscle Growth Media.
  • Cells were then used for experiments or seeded for subculturing at a cell density of ⁇ 3500 cell/cm 2 .
  • Cells were grown in an incubator at 37 0 C and 5% CO 2 and subcultured every 5-6 days.
  • FACS titration on hActRIIB- and hActRIIA-Expressing Cells The half maximal effective concentration (EC50) of the anti-ActRIIB antibodies was determined via binding to cellular hActRIIA and hActRIIB by FACS.
  • serial dilutions of anti-ActRIIB Fab or IgG were incubated with IxIO 4 hActRIIA- transfected, hActRIIB -transfected or parental HEK293T/17 cells per well for 1 h at 4 0 C. After several washing steps cell-bound Fabs or IgGs were detected with phycoerythrin-conjugated goat anti-human IgG (H+L) secondary antibody. After one hour incubation at 4 0 C, the cells were washed again and resuspended in FACS buffer and fluorescence intensity of the cells was determined in a FACSArrayTM instrument.
  • Anti-ActRIIB Fab or IgG as well as isotype control Fab or IgG (lO ⁇ g) were incubated with 10 5 huSkMC in FACS buffer (PBS, 2% FCS, ImM EDTA) per tube for 1 h at 4 0 C. After washing steps, cell-bound Fabs or IgGs were detected with phycoerythrin-conjugated goat anti-human IgG (H+L) secondary antibody which had been diluted 1:200 in FACS buffer. After one hour incubation at 4 0 C on a shaker, the cells were washed again and resuspended in FACS buffer and fluorescence intensity of the cells determined in the FACSCaliberTM instrument.
  • EDC-NHS amine coupling chemistry For direct antigen immobilisation standard EDC-NHS amine coupling chemistry was used. CM5 chips (Biacore, Sweden) were coated with approximately 6000 RU human- or mouse- ActRIIB/Fc, or approximately 1500 RU human- ActRIIA/Fc (according to the activity of the antigens) in 10 mM acetate buffer, pH 4.5. For the reference flow cell, a respective amount of HSA was used. Regeneration was done with 5 ⁇ l 1OmM Glycine/HCl buffer pH1.5.
  • the antigens were not immobilized directly, but captured on a CM5 chip, which was modified with an anti-human-Fc antibody (Fc capture kit, GE Healthcare / Biacore).
  • Fc capture kit Fc capture kit, GE Healthcare / Biacore
  • capture antibody was immobilized, but no antigen captured. Regeneration was achieved using 2 injections of 5 ⁇ L 3M MgCl 2 .
  • CK ASSAY Differentiation was initiated 24 hours after seeding by changing cells from GM to serum-free differentiation medium consisting of skeletal muscle basal medium (skBM). Cells were differentiated for 3 days in the absence and presence of myostatin (R&D systems) or other TGF- b proteins and tested antibodies at given concentrations. Cells were washed with PBS and then lysed with Reporter lysis buffer (Promega) and stored till measurement at -8O 0 C. CK activity was measured using the CK (IFCC) reagent (Thermo Electron). The CK reagent was prepared according to the manufacturers instructions.
  • CK reagent was added and absorbance was immediately read at 340 nm for 20 min, reading interval 1 min.
  • CK standard curves were freshly prepared using CK from rabbit muscle (Roche Diagnostics). Protein content was determined using BCA kit.
  • TREATMENT PROTOCOL Control antibody anti-chicken lysozyme-hlgG
  • anti-human ActRIIB antibodies anti-ActRIIB-MOR8159 and MOR8213, MgG,
  • IgG control i.p. (anti-chicken lysozyme IgG), 25 mg/kg
  • Animals were housed in groups of four to five animals at 25 0 C with a 12:12 h light-dark cycle. They were fed a standard laboratory diet containing 18.2% protein and 3.0% fat with energy of 15.8 MJ/kg (NAFAG 3890, Kliba). Food and water were provided ad libitum. Animal experimentation was carried out according to the regulations effective in the Canton of Basel- City, Switzerland.
  • Results are expressed as mean +/-SEM. Statistical analysis was carried out using Dunnett's multiple comparison test following one-way analysis of variance. Treatment (anti-ActRIIB antibodies MOR8159 and MOR8213 were tested for difference to control (control antibody) and differences were considered to be significant when the probability value was ⁇ 0.05: *: P ⁇ 0.05, **: P ⁇ 0.01, NS: no significance versus IgG control. Statistical analyses were performed by GraphPad Prism version 5.0 (GraphPad Software, Inc). Body weight were calculated by subtracting body weight at day 0, and muscle weight was normalized by body weight at day 0 (initial body weight). Pannings, antibody identification and characterization
  • HuCAL GOLD® library is a Fab library (Knappik et al., 2000) in which all six CDRs are diversified by appropriate methods, and which employs the CysDisplayTM technology for linking the Fab to the phage surface (WO01/05950).
  • HuCAL GOLD ® phagemid library (Rothe et al., 2008) was used to select specific Fab antibody fragments.
  • HuCAL GOLD ® antibody-phages were divided into several pools comprising different VH master genes. These pools were individually subjected to differential cell pannings whereby selection rounds on transiently human ActRIIB transfected cells alternated with selection rounds on recombinant human ActRIIB/Fc protein.
  • phage particles diluted in PBS were mixed with an equal volume of PBS/BSA and blocked.
  • 1x10 7 of the respective hActRIIB expressing cells per phage pool were resuspended in PBS/ 3% FCS/ 0.04% NaN 3 and blocked for one hour at 4 0 C on a shaker. The blocked cells were spun down, resuspended in the pre-blocked phage particles and incubated for three hours.
  • 1x10 7 hActRIIB knock-down cells per phage pool were prepared. The phage-cell complexes were washed in PBS/BSA, followed by washing in PBS. Elution of phage particles from the hActRIIB expressing cells was performed by acidic elution with glycine buffer, pH 2.2. After centrifugation, the eluate was neutralized by adding unbuffered Tris.
  • HuCAL GOLD ® phage Prior to the selections, HuCAL GOLD ® phage were pre-adsorbed in blocking buffer. The blocked phage were added to the coated antigen and incubated for 2 hrs at room temperature. Unspecific phage were washed off with PBST and PBS. Bound phage were eluted by addition of 20 mM DTT. The eluates were used for infection of an E. coli TG-I culture. After infection, the bacteria were plated onto LB/CAM/Glc agar plates and incubated overnight at 37 0 C. The next morning, the colonies were scraped off the plates and the phage were rescued and amplified.
  • the most successful panning approach proved to be differential cell/protein pannings with first panning round on ActRIIB transfected HEK293T/17 cells followed by selection round on recombinant human ActRIIB/Fc and again on transfected cells.
  • Selected Fabs were analyzed for binding to parental or rhActRIIB transfected HEK293 cells.
  • MOR07079 Fab binds preferentially to ActRIIB -transfected cells with an EC50 of 2OnM ( Figure 1).
  • MOR07079 Fab showed inhibitory activity and blocked rhActRIIB/Fc binding to myostatin. Strong myostatin binding inhibition in ELISA was reflected by myostatin inhibition in the reporter gene assay using HEK293-CAGA12 for
  • variable domain fragments of heavy (VH) and light chains (VL) were subcloned from the pMORPH ® X9_FH Fab expression vectors into the pMORPH ® 2_h_Ig vector series for human IgG2. Selected clones were also converted into the silent IgGlLALA format in which leucines at positions 234 and 235 were mutated to alanines to abrogate FcR ⁇ binding and attenuate effector functions.
  • Eukaryotic HKBIl cells were transfected with IgG heavy and light chain expression vector DNA.
  • Cell culture supernatant was harvested at 3 or 7 days post transfection and subjected to standard protein A affinity chromatography. If not otherwise stated, buffer exchange was performed to 1 x Dulbecco's PBS (pH 7.2) and samples were sterile filtered (0.2 ⁇ m).
  • CDR-L3 and CDR- H2 regions were optimized in parallel by cassette mutagenesis using trinucleotide directed mutagenesis (Virnekas et al., 1994, Nucleic Acids Res. 22:5600-5607), whereby the framework regions were kept constant (Nagy et al., 2002, Nature Medicine, 8:801-807).
  • all parental Fab fragments were transferred from the expression vector pMORPH ® X9 into the CysDisplayTM maturation vector pMORPH ® 25 via the Xbal/EcoRI restriction sites.
  • This vector provides the phage protein pill fused N-terminally to a cysteine residue as well as a C-terminal cysteine fused to the Fd antibody chain and thus allows disulfide- linked display of the respective Fab fragments on the phage surface.
  • the CDR-H2 region of each parental Fab was excised and replaced by the highly diversified CDR-H2 maturation cassette.
  • the sizes of the maturation libraries ranged from 4xlO 5 to IxIO 8 clones.
  • the vector background was below 1% in all cases.
  • the quality control by sequencing of single clones revealed a high quality of each library.
  • the antibody-displaying phages from the following maturation libraries were subjected to separate pannings and screenings:
  • biotinylated recombinant hActRIIB/Fc was captured onto Streptavidin-coated Dynabeads.
  • the following protocol was applied: for each phage pool, Streptavidin beads were washed with PBS and resuspended in blocking buffer. Phage particles diluted in PBS were mixed blocking buffer containing 0.1% Tween20 and kept on a rotating wheel. Preclearing of phage particles for removal of Streptavidin- or bead-binding phages was performed twice: Per phage pool blocked Streptavidin beads were added to the blocked phage particles and incubated on a rotating wheel. After separation of the beads via a magnetic device the phage supernatant was transferred to a fresh, pre -blocked reaction tube and pre- adsorption was repeated.
  • the biotinylated hActRIIB/Fc antigen was added to the precleared and blocked phage particles and incubated on a rotating wheel.
  • the phage-antigen complexes were captured using blocked streptavidin beads, added to the phage panning pools and incubated further. Phage particles bound to the streptavidin beads were collected. Beads were then washed with PBST and PBS. Elution of phage particles from the streptavidin beads was performed by addition of 20 mM DTT. The eluate was collected and used for infection of an E. coli TG-I culture grown to an OD ⁇ oonm of 0.6-0.8.
  • the bacterial pellets were resuspended in 2xYT medium, plated onto LB/CAM/Glc agar plates and incubated overnight at 37 0 C. The next morning, the colonies were scraped off the plates and the phages were rescued and amplified mainly as described (Krebs et al., 2001) with the exception that helper phage infected cells were grown at 22 0 C over night in medium containing 0.25 mM IPTG.
  • the third rounds of the solution pannings on biotinylated hActRIIB/Fc were performed according to the protocol of the first round except for decreasing amounts of antigen used and increased stringency of the washing conditions.
  • phage particles diluted in PBS were mixed with an equal volume of PBS/BSA and blocked.
  • phage particles were blocked with PBS/FCS/0.02% NaN 3 at 4 0 C.
  • the blocked cells were spun down, resuspended together with the pre -blocked phage particles and incubated further.
  • the phage-cell complexes were washed with PBS/BSA, followed by washing in PBS. Cells were centrifuged at 410 x g for 2 min at 4 0 C. Acidic elution of phage particles from the hActRIIB expressing huSkMC was performed by a 10 min incubation step with glycine buffer, pH 2.2. After centrifugation, the eluate was neutralized by adding unbuffered Tris. The phage containing supernatant was used for infection of an E. coli TG-I culture grown to an OD ⁇ oonm of 0.6-0.8.
  • the bacterial pellets were resuspended in 2xYT medium, plated onto LB/CAM/Glc agar plates and incubated overnight at 37 0 C. The next morning, the colonies were scraped off the plates and the phages were rescued and amplified mainly as described (Krebs et al., 2001) with the exception that helper phage infected cells were grown at 22 0 C over night in medium containing 0.25 mM IPTG.
  • Fabs were selected for expression and purification, and the most promising further characterized.
  • the Fab clone MOR08067 exhibited good inhibition in the myostatin induced RGA as well as binding to rhActRIIB transfected HEK293 cells. Affinity determination by Biacore revealed KD values to human and mouse ActRIIB/Fc below 100 pM. MOR08067 and other candidates were selected for further optimization by a cross cloning approach, while MOR08067, containing a potential N-linked glycolsylation site was also subjected to a deglycosylation approach.
  • this antibody contained a potential N-linked glycosylation site within the CDR-H2 of the heavy chain. This site was removed to yield MOR08156 and MOR08159. The characterization of these MOR08067 -derivatives is described below. b) Cross Cloning of Optimized Fabs
  • IgG2 expression was performed by transient transfection of HKBIl cells and the full length immunoglobulins were purified from the cell culture supernatants. Upon conversion to IgG, all candidates retained their ability to dose-dependently inhibit myostatin-induced activity in the reporter gene assay (Table 3).
  • Table 3 IC50 determination of anti-ActRIIB IgGs in myo statin-induced luciferase reporter gene assay
  • MOR08159 and MOR08213 were tested for their ability to bind to human primary muscle myoblasts by FACS, and specific binding to those cells was reported, in line with low expression of ActRIIB on those cells ( Figure 4).
  • MOR08159 and MOR08213 displayed ability to fully reverse the myostatin-induced inhibition of primary skeletal myoblasts differentiation (Figure 5). Those antibodies also increased differentiation above basal level in the absence of exogenous myostatin, due to their ability to neutralize endogenously produced ActRIIB ligands.
  • CDR-Ll and CDR-H2 regions were optimized by cassette mutagenesis using trinucleotide directed mutagenesis (Virnekas et al. [supra]), whereby the framework regions were kept constant (Nagy et al. [supra]).
  • all parental Fab fragments were transferred from the expression vector pMORPH ® X9 into the CysDisplayTM maturation vector pMORPH ® 25 via the Xbal/EcoRI restriction sites.
  • Differential pannings for the second round of affinity maturation included parental HEK293T/17 cells and huSkMC expressing human ActRIIB endogeneously at low levels. Additionally, recombinant biotinylated hActRIIB/Fc antigen was included in all panning strategies.
  • bacterial lysates were screened using parental HEK293T/17 and/or hActRIIB-transfected HEK293T/17 cells.
  • Cell-bound Fabs were detected with phycoerythrin- conjugated goat anti-human IgG (H+L) secondary antibody. The quantification of Fab expression in the lysates was performed in parallel.
  • IgG2 conversion was performed by transient transfection of HKBIl cells and the full length immunoglobulins were purified from the cell culture supernatants.
  • Table 4 IC50 determination of anti-ActRIIB IgGs in myostatin-induced inhibition of skeletal muscle differentiation assay
  • TGF ⁇ family IC 50 J ⁇ max IC 50 J ⁇ max IC 50 J ⁇ max ligands ng/ml (% control) (ng/ml) (% control) (ng/ml) (% control)
  • Myo statin 8.5 + 0.6 25 .7 + 1 .5 42 7 + 5 8 28.9 + 4 8 35 1 + 5 1 35.3 + 4 5
  • Table 5 IC50 and Emax determination of various ligand- induced inhibition of skeletal muscle differentiation assay in the presence or absence of MOR08159/MOR08213 (lO ⁇ g/ml)
  • Myostatin and GDF-Il are able to inhibit human myoblasts differentiation with similar efficiencies and to similar extents.
  • MOR08213 myostatin and GDF-Il dose responses were shifted in a parallel manner. Activin A was also able to inhibit differentiation, however in the presence of MOR08159 or MOR08213, we observed a non parallel shift accompanied by a change in Emax and potency. BMP-2 response was unaffected by the presence of MOR08159 or MOR08213, suggesting that it does not occur via ActRIIB binding.
  • MOR08213 was able to induce a dose-dependent profound hypertrophy of all examined muscles at 5 and 25 mg/kg while no significant changes were noticed at lmg/kg dose (Figure 7).
  • Stable human ActRIIB -transfected HEK293T/17 cells were maintained in DMEM containing 10% FBS, 2 mM L-glutamine, penicillin (50 IE/ml), streptomycin (50 ⁇ g/ml) and puromycin (2 ⁇ g/ml). Cells were grown in an incubator at 37 0 C and 5% CO 2 and subcultured every 3-4 days. Cells were detached using AccutaseTM and then transferred into a new flask containing fresh media. The ability of the anti-ActRIIB antibodies to bind to the same epitope of human ActRIIB was assessed by FACS using hActRIIB-expressing cells.
  • an anti-ActRIIB IgG was incubated with IxIO 5 hActRIIB-transfected cells per well for 1 h at 4 0 C. After washing, a different biotinylated anti-ActRIIB IgG or a control biotinylated IgG were incubated at equimolar concentration to the first anti-ActRIIB IgG for 1 h at 4 0 C. After washing, cell -bound biotonylated IgG were detected with streptavidin-APC (Biolegend). After one hour incubation at 4 0 C, the cells were washed again and resuspended in FACS buffer and fluorescence intensity of the cells was determined in a FACSArrayTM instrument.
  • MOR08159 and MOR08213 were tested for their ability to jointly bind to human ActRIIB- transfected cells by FACS, and specific binding of MOR08159 alone (bold black) or in the presence of MOR08213 (bold dashed) was reported compared to isotype control (black) or isotype control in the presence of MOR08213 (dashed) ( Figure 8).
  • MOR08213 In the presence of MOR08213, binding of MOR08159 is significantly reduced suggesting that those two antibodies either bind to the same sites or to sites that may have some degree of overlap or that binding of MOR08213 to a distinct, but nearby site, might sterically hinder binding of MOR08159.
  • a collection of antigen derived cyclic peptides displayed on peptide microarrays were incubated with antibodies of interest.
  • the determination of peptide-antibody binding was performed by RepliTope-analysis where the peptide microarray was incubated with the primary antibody followed by a fluorescently labelled secondary antibody directed against the Fc-part of the primary one.
  • the peptide microarrays where dried using a microarray centrifuge and scanned in a high resolution microarray scanning system with appropriate wavelength settings.
  • the microarray is composed of three subarrays, each displaying cyclic peptides derived from ActRIIB (with Cys residue exchanged to Ser), which were scanned (peptide scan format 15/12).
  • cyclic peptides derived from ActRIIB with Cys residue exchanged to Ser
  • peptide scan format 15/12 peptide scan format 15/12.
  • unrelated antibody ACEl 8543, isotope control
  • fluorescently labelled secondary antibody Cy- 5 labelled anti-human IgG
  • the sequence common to these peptides to which MOR08159 is considered to bind is 76GCWLDDFNC84 (SEQ ID NO: 186).
  • a second region with weaker binding characteristics was also identified using this method. This second region has the sequence 49CEGEQDKRLHCYASW63 (SEQ ID NO: 187).
  • Human ActRIIB aa20-120 and aa24-117 were expressed.
  • MOR08159 Fab and Fv regions were expressed and purified (all expression was carried out in E.coli). Using these proteins, four protein complexes were prepared, purified and crystallised (MOR08159Fab- ActRIIB 20-120, MOR08159Fab- ActRIIB 24-117, MOR08159Fv- ActRIIB 20-120, MOR08159Fv-ActRIIB 24-117).
  • this plate was sealed and incubated over night at 4 0 C. After the incubation the antigen-coated High Bind MTP was washed with PBS/0.05%(w/v)Tween 20. Subsequently, the plate was blocked with PBS/5%(w/v)BSA. The washing steps were repeated and 50 ⁇ l/well of the antibody-antigen preparation from the polypropylene MTP was transferred into the antigen-coated High Bind MTP. The High Bind MTP was incubated for 25 min at room temperature.

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Cited By (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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WO2013063536A1 (en) * 2011-10-27 2013-05-02 Acceleron Pharma, Inc. Actriib binding agents and uses thereof
WO2014030683A1 (ja) * 2012-08-21 2014-02-27 国立大学法人九州大学 貧血患者の貧血の要因を検出するためのバイオマーカー
US8703694B2 (en) 2009-06-08 2014-04-22 Acceleron Pharma, Inc. Methods for increasing thermogenic adipocytes
US8703927B2 (en) 2008-08-14 2014-04-22 Acceleron Pharma Inc. Isolated nucleotide sequences encoding GDF traps
US8710016B2 (en) 2009-11-17 2014-04-29 Acceleron Pharma, Inc. ActRIIB proteins and variants and uses therefore relating to utrophin induction for muscular dystrophy therapy
WO2014097116A1 (en) 2012-12-18 2014-06-26 Novartis Ag Stabilized insulin-like growth factor polypeptides
US8765663B2 (en) 2009-01-13 2014-07-01 Acceleron Pharma Inc. Methods for increasing adiponectin
US8765385B2 (en) 2011-10-27 2014-07-01 Ravindra Kumar Method of detection of neutralizing anti-actriib antibodies
US8895016B2 (en) 2006-12-18 2014-11-25 Acceleron Pharma, Inc. Antagonists of activin-actriia and uses for increasing red blood cell levels
WO2015022658A2 (en) 2013-08-14 2015-02-19 Novartis Ag Methods of treating sporadic inclusion body myositis
WO2015049630A1 (en) 2013-10-02 2015-04-09 Novartis Ag Insulin-like growth factor mimetics for use in therapy
CN104540961A (zh) * 2012-06-11 2015-04-22 安姆根公司 双重受体拮抗性抗原结合蛋白及其用途
WO2015087276A1 (en) 2013-12-12 2015-06-18 Novartis Ag A process for preparing a composition of pegylated proteins
WO2015111008A2 (en) 2014-01-27 2015-07-30 Novartis Ag Biomarkers predictive of muscle atrophy, method and use
US9138459B2 (en) 2004-07-23 2015-09-22 Acceleron Pharma Inc. ACTRIIB-FC polynucleotides, polypeptides, and compositions
WO2015162590A1 (en) 2014-04-24 2015-10-29 Novartis Ag Methods of improving or accelerating physical recovery after surgery for hip fracture
US9353356B2 (en) 2007-09-18 2016-05-31 Acceleron Pharma Inc. Activin-actriia antagonists for treating a follicle-stimulating horomone-secreting pituitary tumor
WO2016092439A1 (en) 2014-12-08 2016-06-16 Novartis Ag Myostatin or activin antagonists for the treatment of sarcopenia
US9480742B2 (en) 2005-11-23 2016-11-01 Acceleron Pharma Inc. Method of promoting bone growth by an anti-actriia antibody
US9493556B2 (en) 2010-11-08 2016-11-15 Acceleron Pharma Inc. Actriia binding agents and uses thereof
US9505813B2 (en) 2008-08-14 2016-11-29 Acceleron Pharma Inc. Use of GDF traps to treat anemia
US9526759B2 (en) 2007-02-01 2016-12-27 Acceleron Pharma Inc. Activin-actriia antagonists and uses for treating or preventing breast cancer
US9572865B2 (en) 2005-11-23 2017-02-21 Acceleron Pharma Inc. Activin-actriia antagonists and uses for treating multiple myeloma
WO2017081624A1 (en) 2015-11-11 2017-05-18 Novartis Ag Uses of myostatin antagonists, combinations containing them and uses thereof
WO2017141208A1 (en) 2016-02-17 2017-08-24 Novartis Ag Tgfbeta 2 antibodies
US9745559B2 (en) 2009-06-12 2017-08-29 Acceleron Pharma Inc. Method for decreasing the body fat content in a subject by administering an ActRIIB protein
US9850298B2 (en) 2014-06-13 2017-12-26 Acceleron Pharma Inc. Methods for treating ulcers in thalassemia syndrome with an ActRIIB polypeptide
WO2017221142A1 (en) 2016-06-22 2017-12-28 Novartis Ag Wnt inhibitors for use in the treatment of fibrosis
WO2018116201A1 (en) 2016-12-21 2018-06-28 Novartis Ag Myostatin, activin or activin receptor antagonists for use in treating obesity and related conditions
US10071135B2 (en) 2005-11-23 2018-09-11 Acceleron Pharma Inc. Method of identifying an agent that promotes bone growth or increases bone density
WO2018175460A1 (en) 2017-03-24 2018-09-27 Novartis Ag Methods for preventing and treating heart disease
WO2019003104A1 (en) 2017-06-28 2019-01-03 Novartis Ag METHOD FOR PREVENTING AND TREATING URINARY INCONTINENCE
US10259861B2 (en) 2007-02-02 2019-04-16 Acceleron Pharma Inc. Variants derived from ActRIIB and uses therefor
US10307455B2 (en) 2016-03-10 2019-06-04 Acceleron Pharma Inc. Activin type 2 receptor antibodies
US10882904B2 (en) 2016-01-08 2021-01-05 Scholar Rock, Inc. Methods for inhibiting myostatin activation by administering anti-pro/latent myostatin antibodies
US10913782B2 (en) 2015-04-22 2021-02-09 Biogen Ma Inc. Hybrid ActRIIB ligand trap proteins for treating muscle wasting diseases
WO2021044287A1 (en) 2019-09-03 2021-03-11 Novartis Ag Treatment of liver disease or disorder comprising actrii receptor antagonists
US10946036B2 (en) 2016-06-13 2021-03-16 Scholar Rock, Inc. Use of myostatin inhibitors and combination therapies
US11135291B2 (en) 2014-11-06 2021-10-05 Scholar Rock, Inc. Methods for making and using anti-myostatin antibodies
US11155611B2 (en) 2017-01-06 2021-10-26 Scholar Rock, Inc. Compositions and methods for making and using anti-myostatin antibodies
US11407799B2 (en) 2016-11-29 2022-08-09 Pharmafox Therapeutics Ag Muscle performance improvement compounds comprising C-terminus agrin fragment and an ACTR2B inhibitor
US11439704B2 (en) 2015-09-15 2022-09-13 Scholar Rock, Inc. Methods for using anti-myostatin antibodies
WO2023028606A1 (en) 2021-08-27 2023-03-02 Versanis Bio, Inc. Combination therapies
WO2024044782A1 (en) 2022-08-26 2024-02-29 Versanis Bio, Inc. Actrii antibody fixed unit dose treatments

Families Citing this family (50)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1855694B1 (en) 2005-02-09 2020-12-02 Sarepta Therapeutics, Inc. Antisense composition for treating muscle atrophy
US20100028332A1 (en) * 2006-12-18 2010-02-04 Acceleron Pharma Inc. Antagonists of actriib and uses for increasing red blood cell levels
TWI573802B (zh) 2007-03-06 2017-03-11 安美基公司 變異之活動素受體多肽及其用途
NZ590327A (en) * 2008-06-26 2013-12-20 Acceleron Pharma Inc Methods for dosing an activin-actriia antagonist and monitoring of treated patients
SG171813A1 (en) 2008-11-26 2011-07-28 Amgen Inc Variants of activin iib receptor polypeptides and uses thereof
MY153078A (en) * 2009-04-27 2014-12-31 Novartis Ag Compositions and methods for increasing muscle growth
CA2773494A1 (en) * 2009-09-09 2011-03-17 Acceleron Pharma Inc. Actriib antagonists and dosing and uses thereof
CA2779472C (en) * 2009-11-03 2021-03-16 Acceleron Pharma Inc. The use of a composition comprising an activin type iib receptor polypeptide in the treatment of fatty liver disease
JO3340B1 (ar) 2010-05-26 2019-03-13 Regeneron Pharma مضادات حيوية لـعامل تمايز النمو 8 البشري
WO2012019168A2 (en) 2010-08-06 2012-02-09 Moderna Therapeutics, Inc. Engineered nucleic acids and methods of use thereof
CA2821992A1 (en) 2010-10-01 2012-04-05 Moderna Therapeutics, Inc. Engineered nucleic acids and methods of use thereof
DE12722942T1 (de) 2011-03-31 2021-09-30 Modernatx, Inc. Freisetzung und formulierung von manipulierten nukleinsäuren
US9464124B2 (en) 2011-09-12 2016-10-11 Moderna Therapeutics, Inc. Engineered nucleic acids and methods of use thereof
EP3492109B1 (en) 2011-10-03 2020-03-04 ModernaTX, Inc. Modified nucleosides, nucleotides, and nucleic acids, and uses thereof
US20130085139A1 (en) 2011-10-04 2013-04-04 Royal Holloway And Bedford New College Oligomers
US8871209B2 (en) 2011-11-14 2014-10-28 Regeneron Pharmaceuticals, Inc. Compositions and methods for increasing muscle mass and muscle strength by specifically antagonizing GDF8 and or Activin A
JP2015501844A (ja) 2011-12-16 2015-01-19 モデルナ セラピューティクス インコーポレイテッドＭｏｄｅｒｎａ Ｔｈｅｒａｐｅｕｔｉｃｓ，Ｉｎｃ． 修飾ヌクレオシド、ヌクレオチドおよび核酸組成物
US10039813B2 (en) 2012-02-07 2018-08-07 Massachusetts Institute Of Technology Use of antagonists of ghrelin or ghrelin receptor to prevent or treat stress-sensitive psychiatric illness
US9283287B2 (en) 2012-04-02 2016-03-15 Moderna Therapeutics, Inc. Modified polynucleotides for the production of nuclear proteins
US9572897B2 (en) 2012-04-02 2017-02-21 Modernatx, Inc. Modified polynucleotides for the production of cytoplasmic and cytoskeletal proteins
US9878056B2 (en) 2012-04-02 2018-01-30 Modernatx, Inc. Modified polynucleotides for the production of cosmetic proteins and peptides
EP2833923A4 (en) 2012-04-02 2016-02-24 Moderna Therapeutics Inc MODIFIED POLYNUCLEOTIDES FOR THE PRODUCTION OF PROTEINS
WO2014066486A2 (en) * 2012-10-24 2014-05-01 Celgene Corporation Biomarker for use in treating anemia
RU2678117C2 (ru) 2012-11-02 2019-01-23 Селджин Корпорейшн Антагонисты активина-actrii и их применение для лечения нарушений костной ткани и других нарушений
LT2922554T (lt) 2012-11-26 2022-06-27 Modernatx, Inc. Terminaliai modifikuota rnr
EP2951205B1 (en) 2013-02-01 2021-08-11 Santa Maria Biotherapeutics, Inc. Anti-activin-a compounds for the treatment of ovarian cancer
US9724396B2 (en) 2013-03-15 2017-08-08 Massachusetts Institute Of Technology Use of antagonists of growth hormone or growth hormone receptor to prevent or treat stress-sensitive psychiatric illness
US8980864B2 (en) 2013-03-15 2015-03-17 Moderna Therapeutics, Inc. Compositions and methods of altering cholesterol levels
WO2014172448A2 (en) * 2013-04-17 2014-10-23 Anaptysbio, Inc. Antibodies directed against activin receptor type ii (actrii)
TW201920262A (zh) 2013-07-30 2019-06-01 美商再生元醫藥公司 抗活化素ａ之抗體及其用途
CA2923422C (en) 2013-09-09 2021-09-07 Vtv Therapeutics Llc Use of a ppar-delta agonist for treating muscle atrophy
WO2015048744A2 (en) 2013-09-30 2015-04-02 Moderna Therapeutics, Inc. Polynucleotides encoding immune modulating polypeptides
EP3052521A1 (en) 2013-10-03 2016-08-10 Moderna Therapeutics, Inc. Polynucleotides encoding low density lipoprotein receptor
EP3094751A4 (en) * 2014-01-14 2017-06-07 Santa Maria Biotherapeutics, Inc. Activin inhibitor response prediction and uses for treatment
MA41052A (fr) 2014-10-09 2017-08-15 Celgene Corp Traitement d'une maladie cardiovasculaire à l'aide de pièges de ligands d'actrii
WO2016090035A2 (en) * 2014-12-02 2016-06-09 La Jolla Institute For Allergy And Immunology Modulators of activin and methods for modulating immune responses and t follicular helper cells
HUE062189T2 (hu) 2014-12-03 2023-09-28 Celgene Corp Aktivin-ACTRII-antagonisták és alkalmazások mielodiszpláziás szindróma kezelésére
WO2016138099A1 (en) 2015-02-24 2016-09-01 Massachusetts Institute Of Technology Use of ghrelin or functional ghrelin receptor agonists to prevent and treat stress-sensitive psychiatric illness
MA41795A (fr) 2015-03-18 2018-01-23 Sarepta Therapeutics Inc Exclusion d'un exon induite par des composés antisens dans la myostatine
CN107771081A (zh) 2015-04-15 2018-03-06 瑞泽恩制药公司 用gdf8抑制剂增加力量和功能的方法
EP3359668A4 (en) 2015-10-09 2019-06-05 Sarepta Therapeutics, Inc. COMPOSITIONS AND METHODS OF TREATING DUCHENNE MUSCLE DYSTROPHY AND ASSOCIATED ILLNESSES THEREOF
EP3439741A4 (en) 2016-04-06 2020-05-06 Acceleron Pharma Inc. ALK7 ANTAGONISTS AND USES THEREOF
CN109071647B (zh) * 2016-04-27 2022-11-22 诺华股份有限公司 抗生长分化因子15的抗体及其用途
AU2017340504A1 (en) * 2016-10-05 2019-04-11 Acceleron Pharma, Inc. Compositions and method for treating kidney disease
WO2018162931A1 (en) * 2017-03-10 2018-09-13 Ucl Business Plc Method relating to myostatin pathway inhibition
EA202092064A1 (ru) 2018-03-01 2020-12-21 Ридженерон Фармасьютикалз, Инк. Способы изменения состава тела
WO2020243448A1 (en) * 2019-05-30 2020-12-03 Acceleron Pharma Inc. Actrii-binding proteins and uses thereof
AU2021327238A1 (en) 2020-08-20 2023-04-27 Regeneron Pharmaceuticals, Inc. Methods for preventing and treating cardiac dysfunction and covid-19 with activin a antagonists
WO2023147309A1 (en) 2022-01-25 2023-08-03 Reneo Pharmaceuticals, Inc. Use of ppar-delta agonists in the treatment of disease
WO2024014989A1 (ru) * 2022-07-14 2024-01-18 Общество с ограниченной ответственностью "БИОМЕД-РЕСУРС" Рекомбинантный белок gbd-actriib для увеличения мышечной массы сельскохозяйственных животных и птицы

Citations (103)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4399216A (en) 1980-02-25 1983-08-16 The Trustees Of Columbia University Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US4439196A (en) 1982-03-18 1984-03-27 Merck & Co., Inc. Osmotic drug delivery system
US4447224A (en) 1982-09-20 1984-05-08 Infusaid Corporation Variable flow implantable infusion apparatus
US4447233A (en) 1981-04-10 1984-05-08 Parker-Hannifin Corporation Medication infusion pump
US4475196A (en) 1981-03-06 1984-10-02 Zor Clair G Instrument for locating faults in aircraft passenger reading light and attendant call control system
US4486194A (en) 1983-06-08 1984-12-04 James Ferrara Therapeutic device for administering medicaments through the skin
US4487603A (en) 1982-11-26 1984-12-11 Cordis Corporation Implantable microinfusion pump system
US4522811A (en) 1982-07-08 1985-06-11 Syntex (U.S.A.) Inc. Serial injection of muramyldipeptides and liposomes enhances the anti-infective activity of muramyldipeptides
EP0154316A2 (en) 1984-03-06 1985-09-11 Takeda Chemical Industries, Ltd. Chemically modified lymphokine and production thereof
US4596556A (en) 1985-03-25 1986-06-24 Bioject, Inc. Hypodermic injection apparatus
US4634665A (en) 1980-02-25 1987-01-06 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
WO1987004462A1 (en) 1986-01-23 1987-07-30 Celltech Limited Recombinant dna sequences, vectors containing them and method for the use thereof
US4790824A (en) 1987-06-19 1988-12-13 Bioject, Inc. Non-invasive hypodermic injection device
WO1989001036A1 (en) 1987-07-23 1989-02-09 Celltech Limited Recombinant dna expression vectors
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
EP0322094A1 (en) 1987-10-30 1989-06-28 Delta Biotechnology Limited N-terminal fragments of human serum albumin
EP0338841A1 (en) 1988-04-18 1989-10-25 Celltech Limited Recombinant DNA methods, vectors and host cells
US4881175A (en) 1986-09-02 1989-11-14 Genex Corporation Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
US4946778A (en) 1987-09-21 1990-08-07 Genex Corporation Single polypeptide chain binding molecules
EP0401384A1 (en) 1988-12-22 1990-12-12 Kirin-Amgen, Inc. Chemically modified granulocyte colony stimulating factor
US5013653A (en) 1987-03-20 1991-05-07 Creative Biomolecules, Inc. Product and process for introduction of a hinge region into a fusion protein to facilitate cleavage
US5064413A (en) 1989-11-09 1991-11-12 Bioject, Inc. Needleless hypodermic injection device
US5091513A (en) 1987-05-21 1992-02-25 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
WO1992003918A1 (en) 1990-08-29 1992-03-19 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
EP0486525A1 (en) 1989-08-01 1992-05-27 Cemu Bioteknik Ab CONJUGATES OF STABILIZED PROTEINS OR PEPTIDES.
US5132405A (en) 1987-05-21 1992-07-21 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5179017A (en) 1980-02-25 1993-01-12 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
WO1993012227A1 (en) 1991-12-17 1993-06-24 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5223409A (en) 1988-09-02 1993-06-29 Protein Engineering Corp. Directed evolution of novel binding proteins
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
US5252479A (en) 1991-11-08 1993-10-12 Research Corporation Technologies, Inc. Safe vector for gene therapy
US5258498A (en) 1987-05-21 1993-11-02 Creative Biomolecules, Inc. Polypeptide linkers for production of biosynthetic proteins
US5260203A (en) 1986-09-02 1993-11-09 Enzon, Inc. Single polypeptide chain binding molecules
WO1994004678A1 (en) 1992-08-21 1994-03-03 Casterman Cecile Immunoglobulins devoid of light chains
US5312335A (en) 1989-11-09 1994-05-17 Bioject Inc. Needleless hypodermic injection device
WO1994025585A1 (en) 1993-04-26 1994-11-10 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5374548A (en) 1986-05-02 1994-12-20 Genentech, Inc. Methods and compositions for the attachment of proteins to liposomes using a glycophospholipid anchor
WO1994029351A2 (en) 1993-06-16 1994-12-22 Celltech Limited Antibodies
US5383851A (en) 1992-07-24 1995-01-24 Bioject Inc. Needleless hypodermic injection device
US5399331A (en) 1985-06-26 1995-03-21 The Liposome Company, Inc. Method for protein-liposome coupling
US5416016A (en) 1989-04-03 1995-05-16 Purdue Research Foundation Method for enhancing transmembrane transport of exogenous molecules
US5427908A (en) 1990-05-01 1995-06-27 Affymax Technologies N.V. Recombinant library screening methods
US5476996A (en) 1988-06-14 1995-12-19 Lidak Pharmaceuticals Human immune system in non-human animal
US5476786A (en) 1987-05-21 1995-12-19 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
US5545806A (en) 1990-08-29 1996-08-13 Genpharm International, Inc. Ransgenic non-human animals for producing heterologous antibodies
US5545807A (en) 1988-10-12 1996-08-13 The Babraham Institute Production of antibodies from transgenic animals
WO1997013852A1 (en) 1995-10-10 1997-04-17 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5624821A (en) 1987-03-18 1997-04-29 Scotgen Biopharmaceuticals Incorporated Antibodies with altered effector functions
US5625126A (en) 1990-08-29 1997-04-29 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5633425A (en) 1990-08-29 1997-05-27 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5661016A (en) 1990-08-29 1997-08-26 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5677425A (en) 1987-09-04 1997-10-14 Celltech Therapeutics Limited Recombinant antibody
US5714350A (en) 1992-03-09 1998-02-03 Protein Design Labs, Inc. Increasing antibody affinity by altering glycosylation in the immunoglobulin variable region
WO1998018943A1 (en) 1996-10-28 1998-05-07 Novartis Ag Method for the oligomerisation of peptides
WO1998024884A1 (en) 1996-12-02 1998-06-11 Genpharm International Transgenic non-human animals capable of producing heterologous antibodies
US5789650A (en) 1990-08-29 1998-08-04 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5814318A (en) 1990-08-29 1998-09-29 Genpharm International Inc. Transgenic non-human animals for producing heterologous antibodies
US5831012A (en) 1994-01-14 1998-11-03 Pharmacia & Upjohn Aktiebolag Bacterial receptor structures
US5869046A (en) 1995-04-14 1999-02-09 Genentech, Inc. Altered polypeptides with increased half-life
US5874299A (en) 1990-08-29 1999-02-23 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5877397A (en) 1990-08-29 1999-03-02 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5885793A (en) 1991-12-02 1999-03-23 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
WO1999016873A1 (de) 1997-09-26 1999-04-08 Arne Skerra Anticaline
US5939598A (en) 1990-01-12 1999-08-17 Abgenix, Inc. Method of making transgenic mice lacking endogenous heavy chains
WO1999045962A1 (en) 1998-03-13 1999-09-16 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5969108A (en) 1990-07-10 1999-10-19 Medical Research Council Methods for producing members of specific binding pairs
WO1999054342A1 (en) 1998-04-20 1999-10-28 Pablo Umana Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity
US6075181A (en) 1990-01-12 2000-06-13 Abgenix, Inc. Human antibodies derived from immunized xenomice
WO2000042072A2 (en) 1999-01-15 2000-07-20 Genentech, Inc. Polypeptide variants with altered effector function
US6121022A (en) 1995-04-14 2000-09-19 Genentech, Inc. Altered polypeptides with increased half-life
US6150584A (en) 1990-01-12 2000-11-21 Abgenix, Inc. Human antibodies derived from immunized xenomice
US6162963A (en) 1990-01-12 2000-12-19 Abgenix, Inc. Generation of Xenogenetic antibodies
US6165745A (en) 1992-04-24 2000-12-26 Board Of Regents, The University Of Texas System Recombinant production of immunoglobulin-like domains in prokaryotic cells
US6172197B1 (en) 1991-07-10 2001-01-09 Medical Research Council Methods for producing members of specific binding pairs
WO2001004144A2 (de) 1999-07-13 2001-01-18 Scil Proteins Gmbh Design von beta-faltblatt-proteinen mit spezifischen bindungseigenschaften
WO2001005950A2 (en) 1999-07-20 2001-01-25 Morphosys Ag Methods for displaying (poly)peptides/proteins on bacteriophage particles via disulfide bonds
US6194551B1 (en) 1998-04-02 2001-02-27 Genentech, Inc. Polypeptide variants
WO2001014424A2 (en) 1999-08-24 2001-03-01 Medarex, Inc. Human ctla-4 antibodies and their uses
US6277375B1 (en) 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
EP1176195A1 (en) 1999-04-09 2002-01-30 Kyowa Hakko Kogyo Co., Ltd. Method for controlling the activity of immunologically functional molecule
WO2002043478A2 (en) 2000-11-30 2002-06-06 Medarex, Inc. Transgenic transchromosomal rodents for making human antibodies
WO2002092780A2 (en) 2001-05-17 2002-11-21 Diversa Corporation Novel antigen binding molecules for therapeutic, diagnostic, prophylactic, enzymatic, industrial, and agricultural applications, and methods for generating and screening thereof
WO2003035835A2 (en) 2001-10-25 2003-05-01 Genentech, Inc. Glycoprotein compositions
US20030153043A1 (en) 1997-05-21 2003-08-14 Biovation Limited Method for the production of non-immunogenic proteins
WO2003074679A2 (en) 2002-03-01 2003-09-12 Xencor Antibody optimization
US6656475B1 (en) * 1997-08-01 2003-12-02 The Johns Hopkins University School Of Medicine Growth differentiation factor receptors, agonists and antagonists thereof, and methods of using same
WO2004039841A2 (en) 2002-10-29 2004-05-13 Borean Pharma A/S Trimeric binding proteins for trimeric cytokines
US20040161738A1 (en) 2000-05-26 2004-08-19 Arumugam Muruganandam Single-domain brain-targeting antibody fragments derived from llama antibodies
US20040175756A1 (en) 2001-04-26 2004-09-09 Avidia Research Institute Methods for using combinatorial libraries of monomer domains
US6818418B1 (en) 1998-12-10 2004-11-16 Compound Therapeutics, Inc. Protein scaffolds for antibody mimics and other binding proteins
WO2004106368A1 (de) 2003-05-28 2004-12-09 Scil Proteins Gmbh Generierung künstlicher bindungsproteine auf der grundlage von ubiquitin-proteinen
US20050048512A1 (en) 2001-04-26 2005-03-03 Avidia Research Institute Combinatorial libraries of monomer domains
US20050053973A1 (en) 2001-04-26 2005-03-10 Avidia Research Institute Novel proteins with targeted binding
US6946292B2 (en) 2000-10-06 2005-09-20 Kyowa Hakko Kogyo Co., Ltd. Cells producing antibody compositions with increased antibody dependent cytotoxic activity
EP1297172B1 (en) 2000-06-28 2005-11-09 Glycofi, Inc. Methods for producing modified glycoproteins
US20050255552A1 (en) 2004-01-20 2005-11-17 Kalobios, Inc. Antibody specificity transfer using minimal essential binding determinants
US20050257278A1 (en) * 1997-08-01 2005-11-17 The Johns Hopkins University School Of Medicine Transgenic non-human animals expressing a truncated activin type II receptor
US20060008844A1 (en) 2004-06-17 2006-01-12 Avidia Research Institute c-Met kinase binding proteins
WO2007109668A2 (en) * 2006-03-20 2007-09-27 The Uab Research Foundation Compositions and methods for improving bone mass through modulation of novel receptors of pth and fragments thereof
WO2007146689A2 (en) 2006-06-09 2007-12-21 Novartis Ag Stabilized insulin-like growth factor polypeptides
WO2008097541A2 (en) * 2007-02-02 2008-08-14 Acceleron Pharma Inc. Variants derived from actriib and uses therefor

Family Cites Families (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5444047A (en) * 1994-06-16 1995-08-22 Dipasquale; Gene Treatment of arthritic and post-surgical orthopedic conditions with Insulin-like Growth Factor-I
JP2000515003A (ja) * 1996-05-20 2000-11-14 ノボ ノルディスク アクティーゼルスカブ タンパク質加水分解物を得る方法
US6696260B1 (en) * 1997-08-01 2004-02-24 The Johns Hopkins University School Of Medicine Methods to identify growth differentiation factor (GDF) binding proteins
WO1999006559A1 (en) 1997-08-01 1999-02-11 The Johns Hopkins University School Of Medicine Methods to identify growth differentiation factor (gdf) receptors
US6682736B1 (en) * 1998-12-23 2004-01-27 Abgenix, Inc. Human monoclonal antibodies to CTLA-4
US7060802B1 (en) * 2000-09-18 2006-06-13 The Trustees Of Columbia University In The City Of New York Tumor-associated marker
BRPI0116728B1 (pt) * 2001-01-05 2018-10-30 Abgenix Inc anticorpos para receptor de fator de crescimento i semelhante à insulina
CA2490411A1 (en) * 2002-06-28 2004-01-08 Centocor, Inc. Mammalian epo mimetic ch1 deleted mimetibodies, compositions, methods and uses
MXPA05012965A (es) * 2003-06-02 2006-03-09 Wyeth Corp Uso de inhibidores de miostatina (gdf8) en conjuncion con corticoesteroides para el tratamiento de desordenes neuromusculares y composiciones farmaceuticas para ello.
US7575751B2 (en) * 2004-04-27 2009-08-18 Research Development Foundation Activin-A mutants
DK2332977T3 (en) * 2004-07-23 2016-02-29 Acceleron Pharma Inc ActRII receptor polypeptides
GB0417487D0 (en) * 2004-08-05 2004-09-08 Novartis Ag Organic compound
EP1778275A2 (en) * 2004-08-12 2007-05-02 Wyeth Combination therapy for diabetes, obesity, and cardiovascular diseases using gdf-8 inhibitors
CA2538208A1 (en) * 2005-05-04 2006-11-04 Universite Laval Modulation of myostatin and use thereof in cell transplantation-based treatment of muscle disease
SI1915397T1 (sl) * 2005-08-19 2015-05-29 Wyeth Llc Antagonistična protitelesa proti GDF-8 in uporabe pri zdravljenju ALS in drugih obolenj, povezanih z GDF-8
WO2007027957A2 (en) * 2005-08-31 2007-03-08 Cell Signaling Technology, Inc. Reagents for the detection of protein phosphorylation in leukemia signaling pathways
US8067562B2 (en) * 2005-11-01 2011-11-29 Amgen Inc. Isolated nucleic acid molecule comprising the amino acid sequence of SEQ ID NO:1
AU2006321906C1 (en) 2005-12-06 2014-01-16 Amgen Inc. Uses of myostatin antagonists
US8097596B2 (en) * 2006-06-30 2012-01-17 Lakewood-Amedex, Inc. Compositions and methods for the treatment of muscle wasting
US8309068B2 (en) * 2006-08-03 2012-11-13 Myostin Therapeutics Pty Ltd. Isolated polypeptides and methods of improving muscle strength
TWI573802B (zh) 2007-03-06 2017-03-11 安美基公司 變異之活動素受體多肽及其用途
MY153078A (en) * 2009-04-27 2014-12-31 Novartis Ag Compositions and methods for increasing muscle growth

Patent Citations (124)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4634665A (en) 1980-02-25 1987-01-06 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US5179017A (en) 1980-02-25 1993-01-12 The Trustees Of Columbia University In The City Of New York Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US4399216A (en) 1980-02-25 1983-08-16 The Trustees Of Columbia University Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
US4475196A (en) 1981-03-06 1984-10-02 Zor Clair G Instrument for locating faults in aircraft passenger reading light and attendant call control system
US4447233A (en) 1981-04-10 1984-05-08 Parker-Hannifin Corporation Medication infusion pump
US4439196A (en) 1982-03-18 1984-03-27 Merck & Co., Inc. Osmotic drug delivery system
US4522811A (en) 1982-07-08 1985-06-11 Syntex (U.S.A.) Inc. Serial injection of muramyldipeptides and liposomes enhances the anti-infective activity of muramyldipeptides
US4447224A (en) 1982-09-20 1984-05-08 Infusaid Corporation Variable flow implantable infusion apparatus
US4487603A (en) 1982-11-26 1984-12-11 Cordis Corporation Implantable microinfusion pump system
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4486194A (en) 1983-06-08 1984-12-04 James Ferrara Therapeutic device for administering medicaments through the skin
EP0154316A2 (en) 1984-03-06 1985-09-11 Takeda Chemical Industries, Ltd. Chemically modified lymphokine and production thereof
US4596556A (en) 1985-03-25 1986-06-24 Bioject, Inc. Hypodermic injection apparatus
US5399331A (en) 1985-06-26 1995-03-21 The Liposome Company, Inc. Method for protein-liposome coupling
WO1987004462A1 (en) 1986-01-23 1987-07-30 Celltech Limited Recombinant dna sequences, vectors containing them and method for the use thereof
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
US5374548A (en) 1986-05-02 1994-12-20 Genentech, Inc. Methods and compositions for the attachment of proteins to liposomes using a glycophospholipid anchor
US4881175A (en) 1986-09-02 1989-11-14 Genex Corporation Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides
US5260203A (en) 1986-09-02 1993-11-09 Enzon, Inc. Single polypeptide chain binding molecules
US5455030A (en) 1986-09-02 1995-10-03 Enzon Labs, Inc. Immunotheraphy using single chain polypeptide binding molecules
US5648260A (en) 1987-03-18 1997-07-15 Scotgen Biopharmaceuticals Incorporated DNA encoding antibodies with altered effector functions
US5624821A (en) 1987-03-18 1997-04-29 Scotgen Biopharmaceuticals Incorporated Antibodies with altered effector functions
US5013653A (en) 1987-03-20 1991-05-07 Creative Biomolecules, Inc. Product and process for introduction of a hinge region into a fusion protein to facilitate cleavage
US5482858A (en) 1987-05-21 1996-01-09 Creative Biomolecules, Inc. Polypeptide linkers for production of biosynthetic proteins
US5258498A (en) 1987-05-21 1993-11-02 Creative Biomolecules, Inc. Polypeptide linkers for production of biosynthetic proteins
US5091513A (en) 1987-05-21 1992-02-25 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5476786A (en) 1987-05-21 1995-12-19 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5132405A (en) 1987-05-21 1992-07-21 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US4790824A (en) 1987-06-19 1988-12-13 Bioject, Inc. Non-invasive hypodermic injection device
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
WO1989001036A1 (en) 1987-07-23 1989-02-09 Celltech Limited Recombinant dna expression vectors
US5677425A (en) 1987-09-04 1997-10-14 Celltech Therapeutics Limited Recombinant antibody
US4946778A (en) 1987-09-21 1990-08-07 Genex Corporation Single polypeptide chain binding molecules
EP0322094A1 (en) 1987-10-30 1989-06-28 Delta Biotechnology Limited N-terminal fragments of human serum albumin
EP0338841A1 (en) 1988-04-18 1989-10-25 Celltech Limited Recombinant DNA methods, vectors and host cells
US5698767A (en) 1988-06-14 1997-12-16 Lidak Pharmaceuticals Human immune system in non-human animal
US5476996A (en) 1988-06-14 1995-12-19 Lidak Pharmaceuticals Human immune system in non-human animal
US5571698A (en) 1988-09-02 1996-11-05 Protein Engineering Corporation Directed evolution of novel binding proteins
US5403484A (en) 1988-09-02 1995-04-04 Protein Engineering Corporation Viruses expressing chimeric binding proteins
US5223409A (en) 1988-09-02 1993-06-29 Protein Engineering Corp. Directed evolution of novel binding proteins
US5545807A (en) 1988-10-12 1996-08-13 The Babraham Institute Production of antibodies from transgenic animals
EP0401384A1 (en) 1988-12-22 1990-12-12 Kirin-Amgen, Inc. Chemically modified granulocyte colony stimulating factor
US5693762A (en) 1988-12-28 1997-12-02 Protein Design Labs, Inc. Humanized immunoglobulins
US6180370B1 (en) 1988-12-28 2001-01-30 Protein Design Labs, Inc. Humanized immunoglobulins and methods of making the same
US5585089A (en) 1988-12-28 1996-12-17 Protein Design Labs, Inc. Humanized immunoglobulins
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
US5416016A (en) 1989-04-03 1995-05-16 Purdue Research Foundation Method for enhancing transmembrane transport of exogenous molecules
EP0486525A1 (en) 1989-08-01 1992-05-27 Cemu Bioteknik Ab CONJUGATES OF STABILIZED PROTEINS OR PEPTIDES.
US5312335A (en) 1989-11-09 1994-05-17 Bioject Inc. Needleless hypodermic injection device
US5064413A (en) 1989-11-09 1991-11-12 Bioject, Inc. Needleless hypodermic injection device
US6114598A (en) 1990-01-12 2000-09-05 Abgenix, Inc. Generation of xenogeneic antibodies
US6162963A (en) 1990-01-12 2000-12-19 Abgenix, Inc. Generation of Xenogenetic antibodies
US5939598A (en) 1990-01-12 1999-08-17 Abgenix, Inc. Method of making transgenic mice lacking endogenous heavy chains
US6150584A (en) 1990-01-12 2000-11-21 Abgenix, Inc. Human antibodies derived from immunized xenomice
US6075181A (en) 1990-01-12 2000-06-13 Abgenix, Inc. Human antibodies derived from immunized xenomice
US5580717A (en) 1990-05-01 1996-12-03 Affymax Technologies N.V. Recombinant library screening methods
US5427908A (en) 1990-05-01 1995-06-27 Affymax Technologies N.V. Recombinant library screening methods
US5969108A (en) 1990-07-10 1999-10-19 Medical Research Council Methods for producing members of specific binding pairs
US5661016A (en) 1990-08-29 1997-08-26 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5633425A (en) 1990-08-29 1997-05-27 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5545806A (en) 1990-08-29 1996-08-13 Genpharm International, Inc. Ransgenic non-human animals for producing heterologous antibodies
US5814318A (en) 1990-08-29 1998-09-29 Genpharm International Inc. Transgenic non-human animals for producing heterologous antibodies
US5625126A (en) 1990-08-29 1997-04-29 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
WO1992003918A1 (en) 1990-08-29 1992-03-19 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5569825A (en) 1990-08-29 1996-10-29 Genpharm International Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5877397A (en) 1990-08-29 1999-03-02 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5874299A (en) 1990-08-29 1999-02-23 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5770429A (en) 1990-08-29 1998-06-23 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5789650A (en) 1990-08-29 1998-08-04 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US6172197B1 (en) 1991-07-10 2001-01-09 Medical Research Council Methods for producing members of specific binding pairs
US5252479A (en) 1991-11-08 1993-10-12 Research Corporation Technologies, Inc. Safe vector for gene therapy
US6582915B1 (en) 1991-12-02 2003-06-24 Medical Research Council Production of anti-self bodies from antibody segment repertories and displayed on phage
US6593081B1 (en) 1991-12-02 2003-07-15 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
US6555313B1 (en) 1991-12-02 2003-04-29 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
US6544731B1 (en) 1991-12-02 2003-04-08 Medical Research Council Production of anti-self antibodies from antibody segment repertories and displayed on phage
US5885793A (en) 1991-12-02 1999-03-23 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
US6521404B1 (en) 1991-12-02 2003-02-18 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
WO1993012227A1 (en) 1991-12-17 1993-06-24 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5714350A (en) 1992-03-09 1998-02-03 Protein Design Labs, Inc. Increasing antibody affinity by altering glycosylation in the immunoglobulin variable region
US6350861B1 (en) 1992-03-09 2002-02-26 Protein Design Labs, Inc. Antibodies with increased binding affinity
US6165745A (en) 1992-04-24 2000-12-26 Board Of Regents, The University Of Texas System Recombinant production of immunoglobulin-like domains in prokaryotic cells
US5383851A (en) 1992-07-24 1995-01-24 Bioject Inc. Needleless hypodermic injection device
US5399163A (en) 1992-07-24 1995-03-21 Bioject Inc. Needleless hypodermic injection methods and device
WO1994004678A1 (en) 1992-08-21 1994-03-03 Casterman Cecile Immunoglobulins devoid of light chains
US5759808A (en) 1992-08-21 1998-06-02 Vrije Universiteit Brussel Immunoglobulins devoid of light chains
WO1994025585A1 (en) 1993-04-26 1994-11-10 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
WO1994029351A2 (en) 1993-06-16 1994-12-22 Celltech Limited Antibodies
US5831012A (en) 1994-01-14 1998-11-03 Pharmacia & Upjohn Aktiebolag Bacterial receptor structures
US6121022A (en) 1995-04-14 2000-09-19 Genentech, Inc. Altered polypeptides with increased half-life
US5869046A (en) 1995-04-14 1999-02-09 Genentech, Inc. Altered polypeptides with increased half-life
WO1997013852A1 (en) 1995-10-10 1997-04-17 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
WO1998018943A1 (en) 1996-10-28 1998-05-07 Novartis Ag Method for the oligomerisation of peptides
WO1998024884A1 (en) 1996-12-02 1998-06-11 Genpharm International Transgenic non-human animals capable of producing heterologous antibodies
US6277375B1 (en) 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
US20030153043A1 (en) 1997-05-21 2003-08-14 Biovation Limited Method for the production of non-immunogenic proteins
US6656475B1 (en) * 1997-08-01 2003-12-02 The Johns Hopkins University School Of Medicine Growth differentiation factor receptors, agonists and antagonists thereof, and methods of using same
US20050257278A1 (en) * 1997-08-01 2005-11-17 The Johns Hopkins University School Of Medicine Transgenic non-human animals expressing a truncated activin type II receptor
WO1999016873A1 (de) 1997-09-26 1999-04-08 Arne Skerra Anticaline
WO1999045962A1 (en) 1998-03-13 1999-09-16 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US6194551B1 (en) 1998-04-02 2001-02-27 Genentech, Inc. Polypeptide variants
WO1999054342A1 (en) 1998-04-20 1999-10-28 Pablo Umana Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity
US6818418B1 (en) 1998-12-10 2004-11-16 Compound Therapeutics, Inc. Protein scaffolds for antibody mimics and other binding proteins
WO2000042072A2 (en) 1999-01-15 2000-07-20 Genentech, Inc. Polypeptide variants with altered effector function
EP1176195A1 (en) 1999-04-09 2002-01-30 Kyowa Hakko Kogyo Co., Ltd. Method for controlling the activity of immunologically functional molecule
WO2001004144A2 (de) 1999-07-13 2001-01-18 Scil Proteins Gmbh Design von beta-faltblatt-proteinen mit spezifischen bindungseigenschaften
WO2001005950A2 (en) 1999-07-20 2001-01-25 Morphosys Ag Methods for displaying (poly)peptides/proteins on bacteriophage particles via disulfide bonds
WO2001014424A2 (en) 1999-08-24 2001-03-01 Medarex, Inc. Human ctla-4 antibodies and their uses
US20040161738A1 (en) 2000-05-26 2004-08-19 Arumugam Muruganandam Single-domain brain-targeting antibody fragments derived from llama antibodies
EP1297172B1 (en) 2000-06-28 2005-11-09 Glycofi, Inc. Methods for producing modified glycoproteins
US6946292B2 (en) 2000-10-06 2005-09-20 Kyowa Hakko Kogyo Co., Ltd. Cells producing antibody compositions with increased antibody dependent cytotoxic activity
WO2002043478A2 (en) 2000-11-30 2002-06-06 Medarex, Inc. Transgenic transchromosomal rodents for making human antibodies
US20050048512A1 (en) 2001-04-26 2005-03-03 Avidia Research Institute Combinatorial libraries of monomer domains
US20040175756A1 (en) 2001-04-26 2004-09-09 Avidia Research Institute Methods for using combinatorial libraries of monomer domains
US20050053973A1 (en) 2001-04-26 2005-03-10 Avidia Research Institute Novel proteins with targeted binding
WO2002092780A2 (en) 2001-05-17 2002-11-21 Diversa Corporation Novel antigen binding molecules for therapeutic, diagnostic, prophylactic, enzymatic, industrial, and agricultural applications, and methods for generating and screening thereof
WO2003035835A2 (en) 2001-10-25 2003-05-01 Genentech, Inc. Glycoprotein compositions
WO2003074679A2 (en) 2002-03-01 2003-09-12 Xencor Antibody optimization
WO2004039841A2 (en) 2002-10-29 2004-05-13 Borean Pharma A/S Trimeric binding proteins for trimeric cytokines
WO2004106368A1 (de) 2003-05-28 2004-12-09 Scil Proteins Gmbh Generierung künstlicher bindungsproteine auf der grundlage von ubiquitin-proteinen
US20050255552A1 (en) 2004-01-20 2005-11-17 Kalobios, Inc. Antibody specificity transfer using minimal essential binding determinants
US20060008844A1 (en) 2004-06-17 2006-01-12 Avidia Research Institute c-Met kinase binding proteins
WO2007109668A2 (en) * 2006-03-20 2007-09-27 The Uab Research Foundation Compositions and methods for improving bone mass through modulation of novel receptors of pth and fragments thereof
WO2007146689A2 (en) 2006-06-09 2007-12-21 Novartis Ag Stabilized insulin-like growth factor polypeptides
WO2008097541A2 (en) * 2007-02-02 2008-08-14 Acceleron Pharma Inc. Variants derived from actriib and uses therefor

Non-Patent Citations (86)

* Cited by examiner, † Cited by third party
Title
"Monoclonal Antibodies For Cancer Detection And Therapy", 1985, ACADEMIC PRESS, article "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", pages: 303 - 16
"Sustained and Controlled Release Drug Delivery Systems", 1978, MARCEL DEKKER, INC.
ALLEN, T.M., NAT. REV. CANCER, vol. 2, 2002, pages 750 - 763
AMON ET AL.: "Monoclonal Antibodies And Cancer Therapy", 1985, ALAN R. LISS, INC., article "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", pages: 243 - 56
BERGE, S.M. ET AL., J. PHARM. SCI., vol. 66, 1977, pages 1 - 19
BERKNER, K.L., CURR. TOP. MICROBIOL. IMMUNOL., vol. 158, 1992, pages 39 - 66
BIRD ET AL., SCIENCE, vol. 242, 1988, pages 423 - 426
BOGDANOVICH ET AL., NATURE, vol. 420, 2002, pages 418 - 421
BOSS, M. A.; WOOD, C. R., IMMUNOLOGY TODAY, vol. 6, 1985, pages 12 - 13
BRADLEY ET AL., CELL MOL. LIFE SCI., vol. 65, 2008, pages 2119 - 2124
BRADLEY L ET AL: "Myostatin as a therapeutic target for musculoskeletal disease", CMLS CELLULAR AND MOLECULAR LIFE SCIENCES, BIRKHÄUSER-VERLAG, BA, vol. 65, no. 14, 21 April 2008 (2008-04-21), pages 2119 - 2124, XP019619981, ISSN: 1420-9071 *
BRENNAN ET AL., SCIENCE, vol. 229, 1985, pages 81 - 83
BRISCOE ET AL., AM. J. PHYSIOL., vol. 1233, 1995, pages 134
CHEN, J. ET AL., EMBO J., vol. 12, 1993, pages 821 - 830
CHEN, J. ET AL., INTERNATIONAL IMMUNOLOGY, vol. 5, 1993, pages 647 - 656
CHOI ET AL., NATURE GENETICS, vol. 4, 1993, pages 117 - 123
CHOTHIA ET AL., NATURE, vol. 342, 1989, pages 877 - 883
CORTEZ-RETAMOZO, V. ET AL., INT J CANCER, vol. 89, 2002, pages 456 - 62
COX, J. P. L. ET AL., EUR. J IMMUNOL., vol. 24, 1994, pages 827 - 836
DUMOULIN, M. ET AL., NATURE, vol. 424, 2003, pages 783 - 788
E. MEYERS; W. MILLER, COMPUT. APPL. BIOSCI., vol. 4, 1988, pages 11 - 17
F. AUSUBEL ET AL.: "Current Protocols in Molecular Biology", 1987, GREENE PUBLISHING AND WILEY INTERSCIENCE
FISHWILD, D. ET AL., NATURE BIOTECHNOLOGY, vol. 14, 1996, pages 845 - 851
FRIGUET B ET AL., J. IMMUNOL METHODS, vol. 77, no. 2, 1985, pages 305 - 319
GLENNIE ET AL., J. IMMUNOL., vol. 139, 1987, pages 2367 - 2375
GOEDDEL: "Gene Expression Technology. Methods in Enzymology", vol. 185, 1990, ACADEMIC PRESS
HANEL C ET AL., ANAL BIOCHEM, vol. 339, no. 1, 2005, pages 182 - 184
HARDING, F.; LONBERG, N., ANN. N. Y. ACAD. SCI., vol. 764, 1995, pages 536 - 546
HELLSTROM ET AL.: "Controlled Drug Delivery", 1987, MARCEL DEKKER, INC., article "Antibodies For Drug Delivery", pages: 623 - 53
HUSTON ET AL., PROC. NATL. ACAD. SCI. USA, vol. 85, 1988, pages 5879 - 5883
HUSTON ET AL., PROC. NATL. ACAD. SCI., vol. 85, 1988, pages 5879 - 5883
J.J. KILLION; I.J. FIDLER, IMRNUNOMETHODS, vol. 4, 1994, pages 273
JONES, P. ET AL., NATURE, vol. 321, 1986, pages 522 - 525
K. KEINANEN; M.L. LAUKKANEN, FEBSLETT., vol. 346, 1994, pages 123
KABAT, E. A. ET AL.: "Sequences of Proteins of Immunological Interest", 1991, U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
KARPOVSKY ET AL., J. EXP. MED., vol. 160, 1984, pages 1686
KNAPPIK ET AL., J MOL BIOL, vol. 296, 2000, pages 57 - 86
KOHLER; MILSTEIN, NATURE, vol. 256, 1975, pages 495
KUROIWA ET AL., NATURE BIOTECHNOLOGY, vol. 20, 2002, pages 889 - 894
LAUWEREYS, M. ET AL., EMBO J, vol. 17, 1998, pages 3512 - 3520
LEE SE-JIN ET AL: "Regulation of muscle growth by multiple ligands signaling through activin type II receptors.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 13 DEC 2005 LNKD- PUBMED:16330774, vol. 102, no. 50, 13 December 2005 (2005-12-13), pages 18117 - 18122, XP002591054, ISSN: 0027-8424 *
LIU, MA ET AL., PROC. NATL. ACAD. SCI. USA, vol. 82, 1985, pages 8648
LONBERG ET AL., NATURE, vol. 368, no. 6474, 1994, pages 856 - 859
LONBERG, N.: "Handbook of Experimental Pharmacology", vol. 113, 1994, pages: 49 - 101
LONBERG, N.; HUSZAR, D., INTERN. REV. IMMUNOL., vol. 13, 1995, pages 65 - 93
M. OWAIS ET AL., ANTIMICROB. AGENTS CHERNOTHER., vol. 39, 1995, pages 180
MCCAFFERTY ET AL., NATURE, vol. 348, 1990, pages 552 - 554
MORRISON, S., SCIENCE, vol. 229, 1985, pages 1202
MUZYCZKA, N., CURR. TOP. MICROBIOL. IMMUNOL., vol. 158, 1992, pages 97 - 129
NAGY ET AL., NATURE MEDICINE, vol. 8, 2002, pages 801 - 807
NEEDLEMAN; WUNSCH, J. MOL, BIOL., vol. 48, 1970, pages 444 - 453
P.G. BLOEMAN ET AL., FEBS LETT., vol. 357, 1995, pages 140
PASTAN, I; KREITMAN, R. J., CURR. OPIN. INVESTIG. DRUGS, vol. 3, 2002, pages 1089 - 1091
PAULUS, BEHRING INS. MITT., 1985, pages 118 - 132
PAYNE, G., CANCER CELL, vol. 3, 2003, pages 207 - 212
PIEHLER J ET AL., J IMMUNOL METHODS, vol. 201, no. 2, 1997, pages 189 - 206
PLESCHBERGER, M. ET AL., BIOCONJUGATE CHEM, vol. 14, 2003, pages 440 - 448
QUEEN, C. ET AL., PROC. NATL. ACAD. SCI. U.S.A., vol. 86, 1989, pages 10029 - 10033
R.J. KAUFMAN; P.A. SHARP, MOL. BIOL., vol. 159, 1982, pages 601 - 621
RIECHMANN, L. ET AL., NATURE, vol. 332, 1998, pages 323 - 327
SAITO, G. ET AL., ADV. DRUG DELIV. REV., vol. 55, 2003, pages 199 - 215
SCHREIER ET AL., J. BIOL. CHEM., vol. 269, 1994, pages 9090
SENTER, P.D.; SPRINGER, C.J., ADV. DRUG DELIV. REV., vol. 53, 2001, pages 247 - 264
SHIELDS, R.L. ET AL., J. BIOL. CHEM., vol. 277, 2002, pages 26733 - 26740
SHIELDS, R.L. ET AL., J. BIOL. CHEN., vol. 276, 2001, pages 6591 - 6604
STIJLEMANS, B. ET AL., J BIOL CHEM, vol. 279, 2004, pages 1256 - 1261
T STRACHAN; A P READ: "Human Molecular Genetics", 1996, BIOS SCIENTIFIC PUBLISHERS LTD, article "Gene Therapy and other Molecular Genetic-based Therapeutic Approaches"
TAKEBE, Y. ET AL., MOL. CELL. BIOL., vol. 8, 1988, pages 466 - 472
TAYLOR, L. ET AL., INTERNATIONAL IMMUNOLOGY, 1994, pages 579 - 591
TAYLOR, L. ET AL., NUCLEIC ACIDS RESEARCH, vol. 20, 1992, pages 6287 - 6295
THORPE ET AL.: "Monoclonal Antibodies '84: Biological And Clinical Applications", 1985, article "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", pages: 475 - 506
THORPE ET AL.: "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", INMUNOL. REV., vol. 62, 1982, pages 119 - 58
TOMIZUKA ET AL., PROC. NATL. ACAD. SCI. USA, vol. 97, 2000, pages 722 - 727
TOMLINSON, I. M. ET AL., J. FOL. BIOL., vol. 227, 1992, pages 776 - 798
TRAIL, P.A. ET AL., CANCER IMMUNOL. IMMUNOTHER., vol. 52, 2003, pages 328 - 337
TSUCHIDA ET AL., ENDO J, 2008
TSUCHIDA ET AL., ENDO J, vol. 55, no. 1, 2008, pages 11 - 21
TUAILLON ET AL., J. IMMUNOL., vol. 152, 1994, pages 2912 - 2920
TUAILLON ET AL., PROC. NATL. ACAD. SCI. USA, vol. 94, 1993, pages 3720 - 3724
UMANA ET AL., NAT. BIOTECH., vol. 17, 1999, pages 176 - 180
UMEZAWA ET AL., BIOCHEM. BIOPHYS. RES. COMMUN., vol. 153, 1988, pages 1038
URLAUB; CHASIN, PROC. NATL. ACAD. SCI. USA, vol. 77, 1980, pages 4216 - 4220
V.V. RANADE, J. CLIN PHARMACOL., vol. 29, 1989, pages 685
VIRNEKAS ET AL., NUCLEIC ACIDS RES., vol. 22, 1994, pages 5600 - 5607
WARD ET AL., NATURE, vol. 341, 1989, pages 544 - 546
WERNER; ALZHEIMER, CYTOKINE GROWTH FACTORS REV, vol. 17, no. 3, 2006, pages 157 - 171

Cited By (92)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9138459B2 (en) 2004-07-23 2015-09-22 Acceleron Pharma Inc. ACTRIIB-FC polynucleotides, polypeptides, and compositions
US11129873B2 (en) 2005-11-23 2021-09-28 Acceleron Pharma Inc. Method for promoting bone growth using activin-actriia antagonists
US10071135B2 (en) 2005-11-23 2018-09-11 Acceleron Pharma Inc. Method of identifying an agent that promotes bone growth or increases bone density
US10239940B2 (en) 2005-11-23 2019-03-26 Acceleron Pharma Inc. Method of promoting bone growth by an anti-actriia antibody
US9572865B2 (en) 2005-11-23 2017-02-21 Acceleron Pharma Inc. Activin-actriia antagonists and uses for treating multiple myeloma
US9480742B2 (en) 2005-11-23 2016-11-01 Acceleron Pharma Inc. Method of promoting bone growth by an anti-actriia antibody
US8895016B2 (en) 2006-12-18 2014-11-25 Acceleron Pharma, Inc. Antagonists of activin-actriia and uses for increasing red blood cell levels
US10093707B2 (en) 2006-12-18 2018-10-09 Acceleron Pharma Inc. Antagonists of activin-ActRIIa and uses for increasing red blood cell levels
US9526759B2 (en) 2007-02-01 2016-12-27 Acceleron Pharma Inc. Activin-actriia antagonists and uses for treating or preventing breast cancer
US10259861B2 (en) 2007-02-02 2019-04-16 Acceleron Pharma Inc. Variants derived from ActRIIB and uses therefor
US9353356B2 (en) 2007-09-18 2016-05-31 Acceleron Pharma Inc. Activin-actriia antagonists for treating a follicle-stimulating horomone-secreting pituitary tumor
US10829533B2 (en) 2008-08-14 2020-11-10 Acceleron Pharma Inc. Combined use of GDF traps and erythropoietin receptor activators to increase red blood cell levels
US11155791B2 (en) 2008-08-14 2021-10-26 Acceleron Pharma Inc. Methods for treating anemia in a subject in need thereof
US8703927B2 (en) 2008-08-14 2014-04-22 Acceleron Pharma Inc. Isolated nucleotide sequences encoding GDF traps
US9932379B2 (en) 2008-08-14 2018-04-03 Acceleron Pharma Inc. Isolated nucleotide sequences encoding GDF traps
US9439945B2 (en) 2008-08-14 2016-09-13 Acceleron Pharma Inc. Isolated nucleotide sequences encoding GDF traps
US10889626B2 (en) 2008-08-14 2021-01-12 Acceleron Pharma Inc. Combined use of GDF traps and erythropoietin receptor activators to increase red blood cell levels
US9505813B2 (en) 2008-08-14 2016-11-29 Acceleron Pharma Inc. Use of GDF traps to treat anemia
US11168311B2 (en) 2008-08-14 2021-11-09 Acceleron Pharma Inc. Methods for treating anemia in a subject in need thereof
US10829532B2 (en) 2008-08-14 2020-11-10 Acceleron Pharma Inc. Combined use of gdf traps and erythropoietin receptor activators to increase red blood cell levels
US11162085B2 (en) 2008-08-14 2021-11-02 Acceleron Pharma Inc. Methods for treating anemia in a subject in need thereof
US10689427B2 (en) 2008-08-14 2020-06-23 Acceleron Pharma Inc. Combined use of GDF traps and erythropoietin receptor activators to increase red blood cell levels
US10377996B2 (en) 2008-08-14 2019-08-13 Acceleron Pharma Inc. Methods of identifying ActRIIB variants
US8765663B2 (en) 2009-01-13 2014-07-01 Acceleron Pharma Inc. Methods for increasing adiponectin
US8703694B2 (en) 2009-06-08 2014-04-22 Acceleron Pharma, Inc. Methods for increasing thermogenic adipocytes
US10968282B2 (en) 2009-06-08 2021-04-06 Acceleron Pharma Inc. Methods for screening compounds for increasing thermogenic adipocytes
US9790284B2 (en) 2009-06-08 2017-10-17 Acceleron Pharma Inc. Methods for increasing thermogenic adipocytes
EP3345921A1 (en) * 2009-06-08 2018-07-11 Acceleron Pharma Inc. Use of anti-actriib antibodies for increasing thermogenic adipocytes
US11066654B2 (en) 2009-06-12 2021-07-20 Acceleron Pharma Inc. Methods and compositions for reducing serum lipids
US10358633B2 (en) 2009-06-12 2019-07-23 Acceleron Pharma Inc. Method for producing an ActRIIB-Fc fusion polypeptide
US9745559B2 (en) 2009-06-12 2017-08-29 Acceleron Pharma Inc. Method for decreasing the body fat content in a subject by administering an ActRIIB protein
US9617319B2 (en) 2009-11-17 2017-04-11 Acceleron Pharma Inc. ActRIIB proteins and variants and uses therefore relating to utrophin induction for muscular dystrophy therapy
US8710016B2 (en) 2009-11-17 2014-04-29 Acceleron Pharma, Inc. ActRIIB proteins and variants and uses therefore relating to utrophin induction for muscular dystrophy therapy
US10968262B2 (en) 2009-11-17 2021-04-06 Acceleron Pharma Inc. Methods of increasing sarcolemmal utrophin
US9493556B2 (en) 2010-11-08 2016-11-15 Acceleron Pharma Inc. Actriia binding agents and uses thereof
WO2013006437A1 (en) 2011-07-01 2013-01-10 Novartis Ag Method for treating metabolic disorders
JP2014523902A (ja) * 2011-07-01 2014-09-18 ノバルティス アーゲー 代謝障害を治療するための方法
US9624301B2 (en) 2011-10-27 2017-04-18 Acceleron Pharma Inc. Polynucleotides encoding an actriib antibody
WO2013063536A1 (en) * 2011-10-27 2013-05-02 Acceleron Pharma, Inc. Actriib binding agents and uses thereof
US8765385B2 (en) 2011-10-27 2014-07-01 Ravindra Kumar Method of detection of neutralizing anti-actriib antibodies
EP2859114A4 (en) * 2012-06-11 2016-08-03 Amgen Inc BINDING PROTEINS FOR AN ANTAGONIST ANTIGEN OF A DOUBLE RECEPTOR AND USES THEREOF
EP3498857A1 (en) * 2012-06-11 2019-06-19 Amgen, Inc. Dual receptor antagonistic antigen-binding proteins and uses thereof
AU2013274347B2 (en) * 2012-06-11 2018-03-08 Amgen Inc. Dual receptor antagonistic antigen-binding proteins and uses thereof
CN104540961A (zh) * 2012-06-11 2015-04-22 安姆根公司 双重受体拮抗性抗原结合蛋白及其用途
US10981999B2 (en) 2012-06-11 2021-04-20 Amgen Inc. Dual receptor antagonistic antigen-binding proteins and uses thereof
JP2015525230A (ja) * 2012-06-11 2015-09-03 アムジエン・インコーポレーテツド デュアル受容体アンタゴニスト性抗原結合タンパク質およびその使用
EP3540070A1 (en) * 2012-06-11 2019-09-18 Amgen Inc. Dual receptor antagonistic antigen-binding proteins and uses thereof
US10266598B2 (en) 2012-06-11 2019-04-23 Amgen Inc. Dual receptor antagonistic antigen-binding proteins and uses thereof
US9453080B2 (en) 2012-06-11 2016-09-27 Amgen Inc. Dual receptor antagonistic antigen-binding proteins and uses thereof
WO2014030683A1 (ja) * 2012-08-21 2014-02-27 国立大学法人九州大学 貧血患者の貧血の要因を検出するためのバイオマーカー
JPWO2014030683A1 (ja) * 2012-08-21 2016-07-28 国立大学法人九州大学 貧血患者の貧血の要因を検出するためのバイオマーカー
WO2014097116A1 (en) 2012-12-18 2014-06-26 Novartis Ag Stabilized insulin-like growth factor polypeptides
EP3626735A1 (en) 2012-12-18 2020-03-25 Novartis AG Stabilized insulin-like growth factor polypeptides
WO2015022658A2 (en) 2013-08-14 2015-02-19 Novartis Ag Methods of treating sporadic inclusion body myositis
WO2015022658A3 (en) * 2013-08-14 2015-05-28 Novartis Ag Methods of treating sporadic inclusion body myositis
WO2015049630A1 (en) 2013-10-02 2015-04-09 Novartis Ag Insulin-like growth factor mimetics for use in therapy
WO2015087276A1 (en) 2013-12-12 2015-06-18 Novartis Ag A process for preparing a composition of pegylated proteins
WO2015111008A2 (en) 2014-01-27 2015-07-30 Novartis Ag Biomarkers predictive of muscle atrophy, method and use
WO2015162590A1 (en) 2014-04-24 2015-10-29 Novartis Ag Methods of improving or accelerating physical recovery after surgery for hip fracture
EP3461495A1 (en) 2014-04-24 2019-04-03 Novartis AG Methods of improving or accelerating physical recovery after surgery for hip fracture
US9969806B2 (en) 2014-04-24 2018-05-15 Novartis Ag Administration of bimagrumab for improving muscle mass and function after hip fracture surgery
US10487144B2 (en) 2014-06-13 2019-11-26 Acceleron Pharma Inc. Methods for treating ulcers in a hemoglobinopathy anemia with a soluble actRIIB polypeptide
US9850298B2 (en) 2014-06-13 2017-12-26 Acceleron Pharma Inc. Methods for treating ulcers in thalassemia syndrome with an ActRIIB polypeptide
US11260107B2 (en) 2014-06-13 2022-03-01 Acceleron Pharma Inc. Methods and compositions for treating ulcers
US11925683B2 (en) 2014-11-06 2024-03-12 Scholar Rock, Inc. Compositions for making and using anti-Myostatin antibodies
US11135291B2 (en) 2014-11-06 2021-10-05 Scholar Rock, Inc. Methods for making and using anti-myostatin antibodies
WO2016092439A1 (en) 2014-12-08 2016-06-16 Novartis Ag Myostatin or activin antagonists for the treatment of sarcopenia
US11932677B2 (en) 2015-04-22 2024-03-19 Alivegen Inc. Hybrid ActRIIB ligand trap proteins for treating muscle wasting diseases
US10913782B2 (en) 2015-04-22 2021-02-09 Biogen Ma Inc. Hybrid ActRIIB ligand trap proteins for treating muscle wasting diseases
US11292826B2 (en) 2015-04-22 2022-04-05 Biogen Ma Inc. Hybrid ActRIIB ligand trap proteins for treating muscle wasting diseases
US11439704B2 (en) 2015-09-15 2022-09-13 Scholar Rock, Inc. Methods for using anti-myostatin antibodies
WO2017081624A1 (en) 2015-11-11 2017-05-18 Novartis Ag Uses of myostatin antagonists, combinations containing them and uses thereof
US10882904B2 (en) 2016-01-08 2021-01-05 Scholar Rock, Inc. Methods for inhibiting myostatin activation by administering anti-pro/latent myostatin antibodies
US12006359B2 (en) 2016-01-08 2024-06-11 Scholar Rock, Inc. Methods for altering body composition by administering anti-pro/latent myostatin antibodies
WO2017141208A1 (en) 2016-02-17 2017-08-24 Novartis Ag Tgfbeta 2 antibodies
AU2017230091B2 (en) * 2016-03-10 2022-04-07 Acceleron Pharma Inc. Activin type 2 receptor binding proteins and uses thereof
US11000565B2 (en) 2016-03-10 2021-05-11 Acceleron Pharma Inc. Methods of increasing muscle mass by administration of activin type 2 receptor antibodies
US12042524B2 (en) 2016-03-10 2024-07-23 Acceleron Pharma Inc. Activin type 2 receptor binding proteins methods of making them
EP3426680A4 (en) * 2016-03-10 2020-03-11 Acceleron Pharma Inc. TYPE 2 ACTIVIN RECEPTOR BINDING PROTEINS AND USES THEREOF
US10307455B2 (en) 2016-03-10 2019-06-04 Acceleron Pharma Inc. Activin type 2 receptor antibodies
US10946036B2 (en) 2016-06-13 2021-03-16 Scholar Rock, Inc. Use of myostatin inhibitors and combination therapies
AU2020210134B2 (en) * 2016-06-13 2021-11-04 Scholar Rock, Inc. Use of myostatin inhibitors and combination therapies
WO2017221142A1 (en) 2016-06-22 2017-12-28 Novartis Ag Wnt inhibitors for use in the treatment of fibrosis
US11407799B2 (en) 2016-11-29 2022-08-09 Pharmafox Therapeutics Ag Muscle performance improvement compounds comprising C-terminus agrin fragment and an ACTR2B inhibitor
WO2018116201A1 (en) 2016-12-21 2018-06-28 Novartis Ag Myostatin, activin or activin receptor antagonists for use in treating obesity and related conditions
US11155611B2 (en) 2017-01-06 2021-10-26 Scholar Rock, Inc. Compositions and methods for making and using anti-myostatin antibodies
US11834508B2 (en) 2017-03-24 2023-12-05 Novartis Ag Method of treating structural and/or functional cardiac abnormalities by administering an anti-ActRII receptor antibody
WO2018175460A1 (en) 2017-03-24 2018-09-27 Novartis Ag Methods for preventing and treating heart disease
WO2019003104A1 (en) 2017-06-28 2019-01-03 Novartis Ag METHOD FOR PREVENTING AND TREATING URINARY INCONTINENCE
WO2021044287A1 (en) 2019-09-03 2021-03-11 Novartis Ag Treatment of liver disease or disorder comprising actrii receptor antagonists
WO2023028606A1 (en) 2021-08-27 2023-03-02 Versanis Bio, Inc. Combination therapies
WO2024044782A1 (en) 2022-08-26 2024-02-29 Versanis Bio, Inc. Actrii antibody fixed unit dose treatments

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