EA046146B1 - ANTIBODIES AGAINST SORTILIN AND METHODS OF THEIR APPLICATION - Google Patents

Description

Cross reference to related applications

This application claims priority to US Provisional Application No. 62/698007, filed July 13, 2018, US Provisional Application No. 62/860184, filed June 11, 2019, and US Provisional Application No. 62/868849, filed June 28, 2019. , which are incorporated herein by reference in their entirety.

Statement Concerning Federally Sponsored Research or Development

This invention was supported by Government Grant No. R44AG050363 from the National Institutes of Health. The government has certain rights to this invention.

Submission of a list of sequences as a text file in ASCII format

The contents of the following ASCII text file provided are incorporated herein by reference in their entirety: Sequence Listing Computer Readable Form (CRF) (file name: 735022001940SEQLIST.TXT, creation date: July 10, 2019, size: 153 kb).

Technical field

This invention relates to antibodies against sortilin, and therapeutic uses of such antibodies.

Prior Art

Sortilin is a type I transmembrane protein that acts as both a receptor for several ligands and is involved in sorting selected cargo from the trans-Golgi network (TGN) to late endosomes and lysosomes for degradation. Sortilin contains a large extracellular domain, part of the VPS10 family, homologous to yeast VPS10P, and contains a 10-blade beta-propeller structure and a cysteine-rich 10CC module (Nykjaer, A et al., (2012) Trends Neurosci 35: 261-270; and Zheng, Y et al., (2011) PLoS One 6: e21023). A small portion of sortilin (<5%) may be shed as a result of ADAM10 or gamma secretase activity (Nykjaer, A et al., (2012) Trends Neurosci 35: 261-270; and Willnow, TE et al., (2011) Curr Opin Lipidol 22: 7985).

Sortilin binds the secreted protein progranulin (PGRN) and targets it for lysosomal degradation, thereby negatively regulating extracellular PGRN levels (Hu, F et al. (2010) Neuron 68, 654-667). Accordingly, sortilin deficiency significantly increases plasma PGRN levels in both in vivo mouse models and in vitro human cells (Carrasquillo, M.M et al., (2010) Am J Hum Genet 87, 890-897; Lee, W.C et al. , (2014) Hum Mol Genet 23, 1467-1478). Moreover, the sortilin polymorphism has been shown to be strongly associated with serum PGRN levels in humans (Carrasquillo MM et al., (2010), Am J Hum Genet. 10; 87(6):890-7).

Progranulin (PGRN) is a secreted growth factor-like, trophic, and anti-inflammatory protein that also plays a role as an adipokine involved in diet-induced obesity and insulin resistance (Nguyen DA et al., (2013). Trends in Endocrinology and Metabolism, 24, 597-606). Progranulin deficiency accounts for approximately 25% of all inherited forms of frontotemporal dementia (FTD), an early-onset neurodegenerative disease. Patients with heterozygous loss-of-function mutations in PGRN have about 50% reduced extracellular protein levels and inevitably develop FTD, making PGRN the gene responsible for this disease (Baker, M et al., (2006) Nature 442, 916-919 ; Carecchio M et al., (2011) J Alzheimers Dis 27, 781-790; Cruts, M et al., (2008) Trends Genet 24, 186-194; Galimberti, D et al., (2010) J Alzheimers Dis 19, 171-177). Additionally, mutant PGRN alleles have been identified in patients with Alzheimer's disease (Seelaar, H et al., (2011). Journal of neurology, neurosurgery, and psychiatry 82, 476486). Importantly, PGRN has a protective effect in several disease models with elevated PGRN levels, accelerating behavioral recovery from ischemia (Tao, J et al., (2012) Brain Res 1436, 130-136; Egashira, Y. et al., ( 2013). J Neuroinflammation 10, 105), suppressing locomotor deficits in a model of Parkinson's disease (Van Kampen, J.M et al. (2014). PLoS One 9, e97032), attenuating pathology in a model of amyotrophic lateral sclerosis (Laird, A.S et al., (2010). PLoS One 5, e13368.) and arthritis (Tang, W et al., (2011). Science 332, 478-484) and preventing memory deficits in a model of Alzheimer's disease (Minami, S.S et al. ., (2014). Nat Med 20, 1157-1164).

Sortilin also binds directly to proneurotrophins such as pro-nerve growth factor (pro-NGF), pro-BDNF (pro-brain-derived neurotrophic factor), proneurotrophin-3, etc., which contain a pro-domain and typically are proapoptotic. Such proneurotrophin precursors are released under stress, and sortilin is involved in regulating their release, as well as binding to the host cell and promoting apoptosis in combination with p75NTR (Willnow, TE et al., (2008) Nat Rev Neurosci 9: 899-909; Nykjaer, A et al., Trends Neurosci 35: 261-270; and Nykjaer, A et al., (2004) Nature 427: 843-848; Hiroko Yano et al., (2009) J Neurosci. 29: 14790-14802. Teng H.K., et al., J Neurosci 25:5455-5463(2005). Sortilin also binds directly to p75NTR (Skeldal S et al., (2012) J Biol Chem.; 287:43798). Sortilin also binds to neurotensin in a region partially overlapping with progranulin binding (Quistgaard, EM et al., (2009) Nat Struct Mol Biol 16: 96-98; and Zheng, Y et al., PLoS One 6: e21023). Sortilin also interacts with Trk receptors NTRK1,

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NTRK2, and NTRK3; and can regulate their anterograde axonal transport and signaling (Vaegter, CB et al., (2011) Nat. Neurosci. 14:54-61). Sortilin also interacts with, and regulates, the processing and targeting of amyloid precursor protein and the resulting production of pathological beta-amyloid peptides (Gustafsen C et al., (2013). J Neurosci. 2;33(1):64- 71).

Sortilin has also been shown to bind to apolipoproteins and lipoprotein lipase; thus, deficiency results in decreased release of VLDL (very low density lipoprotein) from the liver and decreased cholesterol (Willnow, TE et al., (2011) Curr Opin Lipidol 22: 79-85; Kjolby, M et al., (2010) Cell Metab 12: 213-223; Nilsson, SK et al., (2007) Biochemistry 46: 3896-3904; Nilsson, SK et al., (2008) J Biol Chem 283: 25920-25927; and Klinger, SC et al. ., (2011) J Cell Sci 124: 1095-1105). Recently, it has been suggested that sortilin is also directly involved in binding to APP (amyloid beta precursor protein) (Gustafsen, C et al., (2013) J. Neurosc. 33:64-71), as well as the APP processing enzyme BACE1 (beta-secretase) (Gustafsen, C et al., (2013) J. Neurosc. 33:64-71; and Finan, GM et al., J Biol Chem 286: 12602-12616). Sortilin also binds to apolipoprotein E (APOE), amyloid beta (A beta) peptide (Carlo, AS et al., (2013) J. Neurosc, 33: 358-370), and PCSK9 (proprotein convertase subtilisin-kexin type 9) (Gustafsen et al., (2014) Cell Metab, 19:310318). Sortilin has also been shown to bind to, and regulate extracellular levels of, PCSK9, which targets low-density lipoprotein receptor for degradation into lysosomes, leading to elevated LDL cholesterol levels (Gustafsen C et al., (2014). Cell Metab. 2014 Feb 4;19(2):310-8).

When present in intracellular vesicles such as endosomes, the amino-terminal extracellular domain of sortilin is directed toward the cavity where the vesicle cargo resides. The carboxy-terminal intracellular/cytoplasmic domain of sortilin, however, binds to a number of adapter proteins that regulate its directed migration from the surface and into intracellular compartments. These include AP2 (a clathrin adapter, to modulate endocytosis from the cell surface), and the retromer/AP1 complex, which modulates trafficking from early endosomes to the Golgi apparatus for recycling; and interaction with GGA family proteins (localized in the Golgi apparatus, containing gamma-ear, binding factor ADP-ribosylation) to move from the Golgi apparatus directly to early endosomes, usually for subsequent degradation by lysosomes. Thus, sortilin can bind to ligands with its luminal domain while at the same time binding to cytoplasmic adapters that target it to determine intracellular fate, such as degradation, for progranulin and other factors.

Through its various interactions with proteins such as progranulin, sortilin and its numerous ligands have been demonstrated to be involved in various diseases, disorders and conditions such as frontotemporal dementia, amyotrophic lateral sclerosis, amyotrophic lateral sclerosis-frontotemporal dementia phenotypes, Alzheimer's disease, Parkinson's disease, depression, neuropsychiatric disorders, vascular dementia, seizures, retinal dystrophy, age-related macular degeneration, glaucoma, traumatic brain injury, aging, seizures, wound healing, stroke, arthritis and atherosclerotic vascular disease.

Accordingly, there is a need for therapeutic antibodies that specifically bind sortilin proteins and block the binding of sortilin to its ligands, such as progranulin, or otherwise modulate the effective concentration of ligands, for the treatment of one or more diseases, disorders and conditions associated with sortilin activity.

All sources referenced in this document, including patents, patent applications and publications, are hereby incorporated by reference in their entirety.

The essence of the invention

This invention generally relates to compositions that contain antibodies, eg, monoclonal, chimeric, humanized antibodies, antibody fragments, etc., that specifically bind human sortilin, and methods of using such compositions.

In some aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HVR-H1 comprising SEQ ID NO: 1, HVR-H2, containing SEQ ID NO: 2 or SEQ ID NO: 3, and HVR-H3 containing SEQ ID NO: 5 or SEQ ID NO: 6; and the light chain variable region comprises HVR-L1 containing any of SEQ ID NO: 8-27, HVR-L2 containing SEQ ID NO: 29 or SEQ ID NO: 30, and HVR-L3 containing SEQ ID NO: 32.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region containing HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2,

- 2 046146 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRVS (SEQ ID NO: 30), and HVR-L3 containing the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLES (SEQ ID NO: 3), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR -H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR -H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR -H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 10), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR -H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 21), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing amino acid sequence

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ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLHSNGYNYLD (SEQ ID NO: 26), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQGLLRSNGYNYLD (SEQ ID NO: 27), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1) , HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variable region comprises HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVRL3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the antibody comprises a heavy chain variable region with HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variable region comprises HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments of any of the foregoing aspects, the anti-sortilin antibody: (a) reduces cell surface levels of sortilin greater than the level of reduction caused by an anti-sortilin antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 56 and a light chain variable region chain containing the sequence SEQ ID NO: 79; (b) increases extracellular levels of progranulin greater than the level of increase caused by an anti-sortilin antibody containing a heavy chain variable region containing the sequence of SEQ ID NO: 56, and a light chain variable region containing the sequence of SEQ ID NO: 79; (c) inhibits the interaction between sortilin and progranulin greater than the level of inhibition caused by an anti-sortilin antibody containing a heavy chain variable region containing the sequence of SEQ ID NO: 56 and a light chain variable region containing the sequence of SEQ ID NO: 79; or (d) provides any combination of (a)-(c). In some embodiments of any of the foregoing aspects, the anti-sortilin antibody: (a) reduces cell surface sortilin levels greater than the level of reduction caused by an anti-sortilin antibody selected from the group consisting of S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, or S60-8; (b) increases extracellular levels of progranulin greater than the level of increase caused by an anti-sortilin antibody selected from the group consisting of S-60-1, S-60-2, S-60-3, S-60-4, S- 60-7, or S-60-8; (c) inhibits the interaction between sortilin and progranulin greater than the level of inhibition caused by an anti-sortilin antibody selected from the group consisting of S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, or S-60-8; or (d) provides any combination of (a)-(c).

Certain aspects of the present invention provide an anti-sortilin antibody with a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-56; and/or a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 57-58, 60-78, or 80.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 58.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 59.

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In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 55 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, an anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 55 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 58.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 77.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 78.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 79.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 80.

In some aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 57.

In certain aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 60.

In certain aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 61.

In certain aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 72.

In some embodiments, which may be combined with any of the preceding aspects, the anti-sortilin antibody of the present invention is of the IgG class, the IgM class, or the IgA class. In certain specific embodiments, the anti-sortilin antibody is of the IgG class and is of the IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments of the present invention, there is provided an anti-sortilin antibody of an IgG1, IgG2, IgG3, or IgG4 isotype, wherein (a) the antibody is an IgG1 or IgG2 isotype and the Fc region contains an amino acid substitution at position P331S, wherein the residue position numbering corresponds to the system EU numbering; (b) the antibody is an IgG1 isotype and the Fc region contains amino acid substitutions at positions L234A, L235A and P331S, wherein the residue position numbering follows the EU numbering system; (c) the antibody is an IgG1, IgG2, or IgG4 isotype and the Fc region contains an amino acid substitution at position N297A, wherein the residue position numbering follows the EU numbering system; or (d) the Fc region contains amino acid substitutions at positions S267E and L328F, where the residue position numbering follows the EU numbering system. In some embodiments, the antibody is an IgG1 isotype and the Fc region contains amino acid substitutions at positions L234A, L235A, and P331S, wherein the residue position numbering follows the EU numbering system.

In some aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 and a light chain comprising the amino acid sequence of SEQ ID NO: 142.

In some aspects, the present invention provides an antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 138 and a light chain comprising the amino acid sequence of SEQ ID NO: 142.

In some embodiments of the present invention, the sortilin protein is a human

- 5 046146 eternal protein. In some embodiments, the sortilin protein is a wild-type protein. In some embodiments, the sortilin protein is a natural variant. In some embodiments, the anti-sortilin antibody of the present invention specifically binds to the human sortilin protein. In some embodiments, which may be combined with any of the preceding embodiments, the antibody is a human antibody, a bispecific antibody, a monoclonal antibody, a multivalent antibody, a conjugate antibody, or a chimeric antibody. In some embodiments, the anti-sortilin antibody of this invention is a monoclonal antibody.

In some embodiments, the anti-sortilin antibody of the present invention is a bispecific antibody that recognizes a first antigen and a second antigen. In some embodiments, the bispecific antibodies of this invention recognize both sortilin and an antigen that facilitates transport across the blood-brain barrier. In some specific embodiments, the first antigen is sortilin and the second antigen is sortilin, transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor (LPR)-related proteins 1 and 2 -1 and 2), diphtheria toxin receptor, CRM197, llama single domain antibody, TMEM 30(A), protein transduction domain, TAT, Syn-B, penetratin, polyarginine peptide, angiopep peptide, basigin, Glut1, CD98hc, or ANG1005.

In some embodiments of the present invention, the anti-sortilin antibody is an antibody fragment that binds to the human sortilin protein. In some embodiments, an anti-sortilin antibody is an antibody fragment that binds to one or more human proteins selected from the group consisting of human sortilin, a natural variant of human sortilin, and a disease variant of human sortilin. In some embodiments, which may be combined with any of the preceding embodiments, the anti-sortilin antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, or scFv fragment.

In some embodiments, the invention provides an anti-sortilin antibody, wherein (a) the anti-sortilin antibody increases extracellular levels of progranulin, decreases cellular levels of sortilin, inhibits the interaction between sortilin and progranulin, or any combination thereof; (b) an anti-sortilin antibody reduces cell surface levels of sortilin, increases extracellular levels of progranulin, inhibits the interaction between sortilin and progranulin, or any combination thereof; (c) an anti-sortilin antibody reduces cell surface sortilin levels, reduces intracellular sortilin levels, reduces total sortilin levels, or any combination thereof; (d) the anti-sortilin antibody induces sortilin degradation, sortilin cleavage, sortilin internalization, decreased sortilin expression, or any combination thereof; (e) anti-sortilin antibody reduces cellular levels of sortilin and inhibits the interaction between sortilin and progranulin; (f) anti-sortilin antibody reduces cellular levels of sortilin and increases cellular levels of progranulin; and/or (g) the anti-sortilin antibody increases the effective concentration of progranulin.

In some embodiments, the invention provides an anti-sortilin antibody, wherein the anti-sortilin antibody reduces cell surface levels of sortilin, increases extracellular levels of progranulin, inhibits the interaction between sortilin and progranulin, or any combination thereof. In some embodiments, the anti-sortilin antibody has a dissociation constant (KD) with human sortilin that is up to 2.5 times lower than that of an anti-sortilin antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 56 and a variable the light chain region containing the sequence SEQ ID NO: 79, wherein the KD is determined by FACS (see, for example, Example 1). In some embodiments, the antibody has a dissociation constant (KD) for human sortilin that is greater than 1-fold to about 2.1-fold lower than that of an anti-sortilin antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO : 56, and a light chain variable region containing the sequence SEQ ID NO: 79, wherein KD is determined by FACS. In some embodiments, the anti-sortilin antibody has a dissociation constant (KD) with human sortilin that ranges from about 1.10E-8 M to about 4.68E-10 M, with the KD determined by FACS (see for example, example 1), or from about 270 to about 2910 pM, with KD determined by biolayer interferometry (see, for example, example 4). In some embodiments, the antibody has a dissociation constant (KD) with human sortilin that ranges from about 5.0E-10 M to about 1.0E-9 M, with the KD determined by FACS, or about 250-500 pM, while KD is determined by the method of interferometry of biolayers.

In some embodiments, which may be combined with any of the preceding embodiments, the anti-sortilin antibody of the present invention (a) reduces cell surface levels of sortilin with a half-maximal effective concentration (EC50) of

- 6 046146 less than 150 pM, when measured by flow cytometry; (b) reduces cell surface sortilin levels by greater than about 50% at 1.25 nM IgG, by greater than about 80% at 0.63 nM IgG, or by greater than about 69% at 150 nM IgG compared to control, when measured by flow cytometry; increases the secretion of progranulin by more than about 1.13 times compared to the control at 0.63 nM IgG, or more than about 1.22 times compared to the control at 50 nM IgG, as measured by standard ELISA; blocks the binding of progranulin to sortilin with a half-maximal effective concentration (EC ₅₀ ) of less than 0.325 nM, when measured by flow cytometry; (e) blocks the binding of progranulin to sortilin by more than about 88% at 50 nM IgG, or by more than about 27.5% at 150 nM IgG, compared to control, as measured by flow cytometry; or (f) provides any combination thereof.

In some embodiments, the anti-sortilin antibody of the invention (a) reduces cell surface levels of sortilin with a half-maximal effective concentration (EC50) of less than 681 pM as measured by flow cytometry; (b) reduces cell surface sortilin levels by greater than about 40% at 1.25 nM IgG, by greater than about 29% at 0.6 nM IgG, or by greater than about 62% at 150 nM IgG compared to control, when measured by flow cytometry; (c) increases the secretion of progranulin by more than about 1.11 times that of the 0.63 nM IgG control, or by more than about 1.75 times that of the 50 nM IgG control, as measured by a standard ELISA; (d) blocks the binding of progranulin to sortilin with a half-maximal effective concentration (EC ₅₀ ) of less than 0.751 nM, as measured by flow cytometry; (e) blocks the binding of progranulin to sortilin by more than about 90% at 50 nM IgG, or by more than about 95% at 150 nM IgG, compared to control, as measured by flow cytometry; or (f) provides any combination thereof.

In some embodiments, the anti-sortilin antibody of the present invention competes for binding to sortilin with an antibody comprising a heavy chain variable domain and a light chain variable domain of the antibody selected from the group consisting of S-60-10, S-60-11, S -60-12, S60-13, S-60-14, S-60-15 [N33 (wt) (wild type)], S-60-15.1 [N33T], S-60-16, S-60- 18, S-60-19, S-60-24, and any combination thereof. In some embodiments, which may be combined with any of the preceding embodiments, an anti-sortilin antibody of the present invention binds substantially the same sortilin epitope as an antibody comprising a heavy chain variable domain and a light chain variable domain of an antibody selected from the group consisting of S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [ N33T], S-60-16, S-60-18, S-60-19 and S-60-24.

In another aspect, this invention relates to an antibody produced by any of the methods described herein. In some aspects, the present invention provides an isolated nucleic acid comprising a nucleic acid sequence encoding an anti-sortilin antibody according to any of the preceding aspects and embodiments. Accordingly, some aspects provide a vector containing a nucleic acid encoding an anti-sortilin antibody, and some aspects provide an isolated host cell containing such a vector. In certain aspects, the present invention provides a method of producing an anti-sortilin antibody that binds to sortilin, comprising culturing such an isolated host cell such that the anti-sortilin antibody is produced. In some embodiments, the method further includes isolating an antibody produced by the cell. In some aspects, this document provides an isolated antibody that binds to sortilin and is produced by the method of this invention.

In another aspect, the present invention provides a pharmaceutical composition comprising any of the anti-sortilin antibodies described herein and a pharmaceutically acceptable carrier. In some embodiments, this invention relates to the use of any of the antibodies or compositions described herein for the preparation or production of a drug. In some embodiments, the invention relates to the use of any of the antibodies or compositions described herein for the preparation or manufacture of a medicament for the treatment of frontotemporal dementia, progressive supranuclear palsy, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, lateral Amyotrophic sclerosis, traumatic brain injury, spinal cord injury, neuropathic pain, dementia, stroke, Parkinson's disease, acute multiple encephalomyelitis, retinal degeneration, age-related macular degeneration, glaucoma, multiple sclerosis, septic shock, bacterial infection, arthritis or osteoarthritis. Some embodiments provide an antibody or composition described herein for use in a method of treatment. Some embodiments provide an antibody or composition described herein for use in a method of treating frontotemporal dementia, progressive supranuclear palsy, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, amyotrophic lateral sclerosis, traumatic brain injury, spinal cord injury brain, neuropathic pain, dementia, stroke, disease

- 7 046146

Parkinson's, acute disseminated encephalomyelitis, retinal degeneration, age-related macular degeneration, glaucoma, multiple sclerosis, septic shock, bacterial infection, arthritis or osteoarthritis.

In certain aspects, this document provides a method of preventing, reducing the risk of, or treating an individual with a disease, disorder, or lesion, comprising administering to the individual in need thereof a therapeutically effective amount of an anti-sortilin antibody of the present invention. In some embodiments, the disease, disorder, or lesion is selected from the group consisting of frontotemporal dementia, progressive supranuclear palsy, Alzheimer's disease, vascular dementia, seizure disorder, retinal dystrophy, amyotrophic lateral sclerosis, traumatic brain injury, spinal cord injury, neuropathic pain, dementia, stroke, Parkinson's disease, acute disseminated encephalomyelitis, retinal degeneration, age-related macular degeneration, glaucoma, multiple sclerosis, septic shock, bacterial infection, arthritis and osteoarthritis. In some embodiments, the disease, disorder, or lesion is selected from frontotemporal dementia, progressive supranuclear palsy, Alzheimer's disease, and amyotrophic lateral sclerosis.

In certain aspects, this disclosure provides a method of inhibiting one or more of neuroinflammation, axonopathy characterized by short axonal sprouting and aberrant branching, microglial activation, and inflammatory response, comprising administering to a subject a therapeutically effective amount of an anti-sortilin antibody of the present invention.

In certain aspects, this disclosure provides a method of enhancing one or more of wound healing, autophagy, and the clearance of aggregated proteins, comprising administering to an individual a therapeutically effective amount of an anti-sortilin antibody of the invention.

In certain aspects, this disclosure provides a method of preventing, reducing the risk of, or treating an individual with arthritis, comprising administering to the individual a therapeutically effective amount of an anti-sortilin antibody of the present invention.

In certain aspects, this disclosure provides a method of reducing the expression of one or more pro-inflammatory mediators, comprising administering to an individual a therapeutically effective amount of an anti-sortilin antibody of the invention. In some embodiments, the one or more proinflammatory mediators are selected from the group consisting of IL6, IL12p70, IL12p40, IL-Τβ, TNF-α, CXCL1, CCL2, CCL3, CCL4, and CCL5.

In some aspects, this document provides a method of inhibiting the interaction between sortilin and progranulin, comprising exposing a cell expressing sortilin to an anti-sortilin antibody or pharmaceutical composition of the present invention. In some embodiments, the cell is in an in vitro environment. In some embodiments, the cell is in an in vivo environment. In some embodiments, the method further includes reducing the level of sortilin expressed on the surface of the cell. In some embodiments, extracellular levels of progranulin are increased.

In some aspects, this disclosure provides a method of increasing progranulin levels in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an anti-sortilin antibody or a pharmaceutical composition of the invention. In some embodiments, progranulin levels are increased in plasma. In some embodiments, progranulin levels are increased in the cerebrospinal fluid. In some aspects, this disclosure provides a method of reducing sortilin levels in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an anti-sortilin antibody or a pharmaceutical composition of the present invention. In some embodiments, sortilin levels are decreased in peripheral blood leukocytes. In some embodiments, the individual has one or more mutations in the gene encoding progranulin. In some embodiments, the individual is heterozygous for one or more loss-of-function mutations in the gene encoding progranulin. In some embodiments, the individual has a hexanucleotide repeat expansion c9orf72 (chromosome 9 open reading frame 72). The individual has, or is at risk of developing, frontotemporal dementia, Alzheimer's disease, or amyotrophic lateral sclerosis.

In some embodiments, which may be combined with any of the preceding aspects or embodiments, the present invention provides a method wherein the anti-sortilin antibody comprises two or more anti-sortilin antibodies.

- 8 046146

Brief description of graphic materials

Fig. 1A-1C show pharmacokinetic and pharmacodynamic studies in non-human primates treated with single doses of anti-sortilin antibodies S-60-15.1 [N33T] LALAPS (S-60-15.1 [N33T] with huIgG1 with L234A/L235A/P331S mutations). In fig. 1A shows SORT1 levels in peripheral blood leukocytes as a percentage of baseline at the indicated time points after dosing (hours) of the indicated anti-sortilin antibody. SORT1 expression is reduced at all doses of anti-sortilin antibody tested. Higher doses of antibody (60 mg/kg, 200 mg/kg) resulted in both an earlier and longer-lasting decline in SORT1 levels compared with lower doses of anti-sortilin antibody (5 mg/kg, 20 mg/kg). In fig. 1B shows plasma PGRN levels as a percentage of baseline at the indicated time points after administration (hours) of the indicated doses of anti-sortilin antibody. PGRN levels increase in a time- and dose-dependent manner. Specifically, plasma PGRN levels increased 3- to 4-fold at C _max compared to baseline levels for all anti-sortilin antibody doses tested and remained elevated for longer periods of time at higher antibody doses. In fig. 1C shows CSF PGRN levels as a percentage of baseline at the indicated time points after administration (hours) of the indicated doses of anti-sortilin antibody. CSF PGRN levels increase 2- to 3-fold over baseline in animals receiving 20 mg/kg, 60 mg/kg, or 200 mg/kg. Similar to plasma PGRN levels (Fig. 1B), CSF PGRN levels remained elevated over time in the higher antibody dose groups. For fig. 1A-1C, n=3 animals per dose.

Fig. 2A-2C show pharmacokinetic and pharmacodynamic studies in non-human primates receiving repeated doses of the anti-sortilin antibody S-60-15.1 [N33T] LALAPS. Animals (2 males and 2 females) were administered the anti-sortilin antibody S-60-15.1 [N33T] LALAPS at a dose of 60 mg/kg once a week for four weeks. Dose days are indicated by vertical dotted lines. In fig. 2A presents mean (+/- standard deviation) SORT1 concentrations in peripheral blood white blood cells (WBC) as a percentage of baseline at the indicated time points (days). SORT1 levels in peripheral blood leukocytes remained reduced throughout the studies. In fig. 2B presents the mean (+/-standard deviation) plasma PGRN concentrations as a percentage of baseline (normalized) at the indicated time points (days). Plasma PGRN levels increased 5- to 6-fold above baseline at peak levels. A decrease in plasma PGRN was observed after the fourth and final administration of anti-sortilin antibody; however, plasma PGRN levels remain elevated 2-fold from baseline. In fig. Figure 2C presents the mean (+/- standard deviation) CSF PGRN concentrations as a percentage of baseline (normalized) at the indicated time points (days). CSF PGRN levels were elevated 34-fold from baseline (Fig. 2C).

Detailed description of the invention

Definitions.

As used herein, the term prevention includes providing prevention against the occurrence or recurrence of a particular disease, disorder or condition in an individual. An individual may be predisposed to, susceptible to, or at risk of developing a particular disease, disorder or condition, but have not yet been diagnosed with the disease, disorder or condition.

As used herein, an individual at risk of developing a particular disease, disorder, or condition may or may not have a detectable disease or symptoms of a disease, and may or may not have detectable manifestations of a disease or symptoms of a disease prior to treatment methods described herein. At risk means that an individual has one or more risk factors, which are measurable parameters that are correlated with the development of a specific disease, disorder or condition, as known to those skilled in the art. An individual with one or more such risk factors has a higher likelihood of developing a particular disease, disorder or condition than an individual without one or more such risk factors.

As used herein, the term treatment refers to a clinical intervention aimed at changing the natural course of events for an individual receiving treatment for a clinical condition. Desirable effects of treatment include a reduction in the rate of progression, improvement or alleviation of a pathological condition, and remission or improved prognosis of a particular disease, disorder or condition. An individual successfully undergoes treatment, for example, if one or more symptoms associated with a particular disease, disorder or condition are alleviated or eliminated.

An effective amount refers to at least an amount that is effective, in dosages and for periods of time, necessary to achieve the desired therapeutic or prophylactic result. An effective amount may be provided in one or more administrations. The effective amount herein may vary depending on factors such as the disease state, age, sex and weight of the individual, and the ability of the treatment to produce the desired response in the individual. An effective amount is also such that

- 9 046146 the therapeutically beneficial effects exceed any toxic or harmful effects of the treatment. For prophylactic use, beneficial or desirable results include such results as eliminating or reducing the risk of, reducing the severity of, or delaying the onset of disease, including biochemical, histological and/or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes observed during the course of the disease. . For therapeutic use, beneficial or desirable results include clinical results such as reducing one or more symptoms caused by a disease, improving the quality of life of individuals suffering from a disease, reducing the dose of other drugs required to treat a disease, enhancing the effect of another drug, e.g. targeted delivery, slowing disease progression, and/or increasing survival. An effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to provide prophylactic or therapeutic treatment, directly or indirectly. As understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, an effective amount may be considered in the context of the administration of one or more therapeutic agents, and one agent may be considered to have been administered in an effective amount if, in combination with one or more other agents, the desired result can be or has been achieved

As used herein, administration in combination with another compound or composition includes simultaneous administration and/or administration at a different time. Administration in combination also includes administration as a joint composition or administration as separate compositions, including different dosing frequencies or intervals, and the use of the same route of administration or different routes of administration.

Individual, for the purposes of treatment, prevention or risk reduction, refers to any animal classified as a mammal, including humans, domesticated, farm and zoo animals, sporting animals or pets such as dogs, horses, rabbits, cattle , pigs, hamsters, gerbils, mice, ferrets, rats, cats, etc. Preferably, the individual is a human.

The terms sortilin or sortilin polypeptide are used interchangeably herein to refer to any native sortilin from any mammalian source, including primates (eg, humans and cynos) and rodents (eg, mice and rats), unless otherwise noted other. In some embodiments, the term includes both wild-type sequences and natural sequence variants, such as splice variants or allelic variants. In some embodiments, the term includes full-length, unprocessed sortilin, as well as any form of sortilin that is produced by cellular processing. In some embodiments, the sortilin is human sortilin. In some embodiments, the amino acid sequence of an exemplary human sortilin is SEQ ID NO: 81.

The terms anti-sortilin antibody, antibody that binds sortilin, and antibody that specifically binds sortilin refer to an antibody that is capable of binding sortilin with sufficient affinity such that the antibody is suitable for use as a diagnostic and/or therapeutic agent when targeting sortilin. In one embodiment, the extent of binding of the anti-sortilin antibody to an unrelated, non-sortilin polypeptide is less than about 10% of the binding of the antibody to sortilin, as measured, for example, by radioimmunoassay (RIA). In some embodiments, the antibody that binds to sortilin has a dissociation constant (KD) of <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (eg 10-8 M or less, eg 10-8 M to 10-13 M, eg 10-9 M to 10-13 M). In some embodiments, the anti-sortilin antibody binds to an epitope of sortilin that is conserved between sortilin from different species.

The term immunoglobulin (Ig) is used interchangeably with antibody herein. The term antibody is used herein in its broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), including those formed by at least two intact antibodies, and antibody fragments, provided they exhibit the desired biological activity .

Native antibodies are typically heterotetrameric glycoproteins of approximately 150,000 Daltons in size, consisting of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to the heavy chain by a single covalent disulfide bond, while the number of disulfide bonds in the heavy chains of different immunoglobulin isotypes varies. Each heavy and light chain also has evenly spaced intrachain disulfide bridges. Each heavy chain has a variable domain ( _VH ) at one end, followed by a number of constant domains. Each light chain has a variable domain

- 10 046146 at one end (VL) and a constant domain at its other end; the light chain constant domain is adjacent to the first heavy chain constant domain, and the light chain variable domain is adjacent to the heavy chain variable domain. Specific amino acid residues are believed to form the interface between the light chain and heavy chain variable domains.

The structure and properties of different classes of antibodies are described, for example, in Basic and Clinical Immunology, 8th Ed., Daniel P. Stites, Abba I. Terr and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, page 71 and chapter 6.

The light chain of any vertebrate species can be classified into one of two distinct types, designated kappa (k) and lambda (λ), based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains (CH), immunoglobulins can be classified into different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, having heavy chains designated alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), respectively. The γ and α classes are further divided into subclasses (isotypes) based on relatively minor differences in CH sequence and function, for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The structures and three-dimensional configurations of the subunits of the various classes of immunoglobulins are well known and described generally, for example, in Abbas et al., Cellular and Molecular Immunology, 4th ed. (W.B. Saunders Co., 2000).

The variable region or variable domain of an antibody, such as the anti-sortilin antibody of the present invention, refers to the amino-terminal domains of the heavy or light chain of the antibody. The heavy chain and light chain variable domains may be designated _VH and VL, respectively. These domains are usually the most variable parts of the antibody (compared to other antibodies of the same class) and contain antigen-binding sites.

The term variable refers to the fact that certain segments of the variable domains differ significantly in sequence between different antibodies, such as the anti-sortilin antibodies of the present invention. The variable domain mediates antigen binding and determines the specificity of a particular antibody for its particular antigen. However, variability is not distributed evenly across the entire extent of the variable domains. Instead, it is concentrated in three segments called hypervariable regions (HVRs) of the variable domains of both the light and heavy chains. The more highly conserved regions of the variable domains are called framework regions (FR). The variable domains of the native heavy and light chains each contain four FR regions, predominantly adopting a beta sheet configuration, connected by three HVRs that form loops connecting, and in some cases forming part of, the beta sheet structure. The HVRs on each strand are held in close proximity to each other by framework regions (FRs) and, together with the HVRs of the other strand, participate in the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health , Bethesda, MD (1991)). Constant domains are not directly involved in the binding of the antibody to the antigen, but exhibit various effector functions, such as the participation of the antibody in antibody-dependent cellular toxicity.

An isolated antibody, such as the anti-sortilin antibody of the present invention, is an antibody that has been identified, isolated and/or recovered from a component of its production environment (eg, naturally or recombinantly). Preferably, the isolated polypeptide is not associated with all other contaminants from its production environment. Contaminants from its production environment, such as those produced by recombinant transfected cells, are materials that will typically interfere with research, diagnostic or therapeutic use of the antibody, and may include enzymes, hormones, and other protein or non-protein solutes. In preferred embodiments, the polypeptide will be purified: (1) to greater than 95 weight percent antibody as determined, for example, by the Lowry method, and in some embodiments, to greater than 99 weight percent antibody; (2) to an extent sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a spin-beaker sequencer, or (3) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie staining blue or preferably silver. The isolated antibody includes the antibody in situ in the recombinant T cells, since at least one component of the antibody's natural environment will be absent. Typically, however, the isolated polypeptide or antibody will be obtained using at least one purification step.

The term monoclonal antibody, as used herein, refers to an antibody, such as the anti-sortilin monoclonal antibody of the present invention, derived from a population of substantially homogeneous antibodies, i.e. the individual antibodies that make up the population are identical, except for possible natural mutations and/or post-translational modifications (e.g., isomerization, amidation, etc.) that may be present in

- 11 046146 significant quantities. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Unlike polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibodies have the advantage that they are synthesized by a hybridoma culture uncontaminated by other immunoglobulins. The definition of monoclonal refers to the nature of the antibody as being derived from an essentially homogeneous population of antibodies, and does not imply a requirement that the antibody be produced by any particular method. For example, monoclonal antibodies for use in accordance with this invention can be prepared by various methods, including, without limitation, one or more of the following methods - methods of immunization of animals including, without limitation, rats, mice, rabbits, guinea pigs, hamsters and/or chickens, one or more of DNA, virus-like particles, polypeptide(s), and/or cell(s), hybridoma techniques, B cell cloning techniques, recombinant DNA techniques, and human or humanoid antibody technologies in animals having part or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences.

The terms full-length antibody, intact antibody, or whole antibody are used interchangeably to refer to an antibody, such as the anti-sortilin antibody of the present invention, in its substantially intact form, as opposed to an antibody fragment. More specifically, whole antibodies include heavy and light chain antibodies, including the Fc region. The constant domains may be native sequence constant domains (eg, human native sequence constant domains) or amino acid sequence variants thereof. In some cases, an intact antibody may have one or more effector functions.

The antibody fragment contains a portion of the intact antibody, preferably the antigen binding and/or variable region of the intact antibody. Examples of antibody fragments include Fab-, Fab'-, F(ab')2. and Fv fragments; diabodies; linear antibodies (see US patent 5641870, example 2; Zapata et al., Protein Eng. 8(10): 1057-1062 (1995)); single-chain antibody molecules and multispecific antibodies formed from antibody fragments.

Papain hydrolysis of antibodies, such as the anti-sortilin antibodies of this invention, produces two identical antigen binding fragments, called Fab'' fragments, and a residual Fc fragment, the name of which reflects the ability to readily crystallize. The Fab fragment consists of a light (L) chain together with a heavy (H) chain variable domain (VH), and the first constant domain of a single heavy chain (CH1). Each Fab fragment is monovalent in antigen binding, i.e. it has one antigen-binding site. Treatment of the antibody with pepsin produces one large F(ab') ₂ fragment, which corresponds to approximately two disulfide-linked Fab fragments that have different antigen-binding activities and retain the ability to cross-link antigen. Fab' fragments differ from Fab fragments by the presence of several additional residues at the carboxyl terminus of the CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is used herein to refer to Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab') ₂ antibody fragments were initially obtained in the form of pairs of Fab' fragments, between which hinge cysteines are located. Other chemical combinations of antibody fragments are also known.

The Fc fragment contains the carboxy-terminal portions of both H chains, held together by disulfide bonds. The effector functions of antibodies are determined by sequences in the Fc region, a region that is also recognized by Fc receptors (FcRs) present on certain cell types.

Fv is a minimal antibody fragment that contains a complete antigen recognition and binding site. This fragment consists of a dimer of variable region domains of one heavy and one light chain with strong non-covalent bonds. When these two domains are folded, six hypervariable loops are formed (3 loops in each of the H- and L-chains), the amino acid residues of which are involved in antigen binding and give the antibody antigen-binding specificity. However, even a single variable domain (or half of an Fv containing only three antigen-specific HVRs) has the ability to recognize and bind antigen, although with lower affinity than the entire binding site.

Single chain Fv, also abbreviated as sFv or scFv, refers to antibody fragments that contain the VH and VL domains of the antibody linked into a single polypeptide chain. Preferably, the sFv polypeptide further comprises a polypeptide linker between the VH and _VL domains, which allows the sFv to form a structure desired for antigen binding. sFv is reviewed by Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

Functional antibody fragments, such as the anti-sortilin antibodies of the present invention, contain a portion of an intact antibody, typically including an antigen binding or variable region of an intact antibody or an F region of an antibody that retains or has modified FcR binding ability. Examples of antibody fragments include linear antibody, single chain antibody molecules, and multispecific antibodies formed from antibody fragments.

The term ''diabodies'' refers to small antibody fragments made by constructing sFv fragments (see previous paragraph) with short linkers (approximately 5-10 residues) between the VH and VL domains so as to allow interchain but not intrachain pairing variable domains, resulting in the formation of a bivalent fragment, i.e. fragment having two antigen-binding sites. Bispecific diabodies are heterodimers of two 'crossover' sFv fragments, in which the VH and VL domains of the two antibodies are located in different polypeptide chains.

As used herein, a chimeric antibody refers to an antibody (immunoglobulin), such as the anti-sortilin chimeric antibody of this invention, in which a portion of the heavy and/or light chain is identical or homologous to the corresponding sequence of antibodies derived from or belonging to a particular species class or subclass of antibodies, while the remaining chain(s) are identical or homologous to the corresponding sequences of antibodies derived from another species or belonging to another class or subclass of antibodies, as well as fragments of such antibodies, provided that they exhibit the desired biological activity. Chimeric antibodies of interest to the present invention include PRIMATIZED® antibodies, in which the antigen binding region of the antibody is derived from an antibody produced by, for example, immunizing macaques with the antigen of interest. As used herein, a humanized antibody is used a subset of chimeric antibodies.

Humanized forms of non-human (eg, murine) antibodies, such as the humanized forms of anti-sortilin antibodies of the present invention, are chimeric antibodies containing amino acid residues from non-human HVRs and amino acid residues from human FRs. In some embodiments, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains in which all or substantially all of the HVRs (e.g., CDRs) correspond to the non-human antibody, and all or substantially all of the FRs correspond to a human antibody. The humanized antibody may optionally contain at least a portion of an antibody constant region derived from a human antibody. A humanized form of an antibody, for example a non-human antibody, refers to an antibody that has been humanized.

A human antibody is an antibody having the amino acid sequence corresponding to an antibody, such as the anti-sortilin antibody of the present invention, produced by a human and/or prepared using any methods for producing human antibodies as disclosed herein. This definition of a human antibody expressly excludes a humanized antibody containing non-human antigen-binding residues. Human antibodies can be produced using a variety of methods known in the art, including phage display libraries and yeast platform technologies. Human antibodies can be produced by administering an antigen to a transgenic animal that has been modified to produce such antibodies in response to an antigenic stimulus but with the endogenous loci blocked, for example in immunized xenomice, and also through human B-cell hybridoma technology.

The term hypervariable region, HVR, or HV, as used herein, refers to regions of the variable domain of an antibody, such as the anti-sortilin antibodies of the present invention, that are sequence hypervariable and/or form structurally defined loops. Typically, antibodies contain six HVRs; three in VH (H1, H2, H3), and three in VL (L1, L2, L3). In native antibodies, H3 and L3 show the greatest diversity of the six HVRs, and H3 in particular is thought to play a unique role in conferring fine specificity to antibodies. Natural camelid antibodies, consisting only of the heavy chain, are functional and stable in the absence of the light chain.

A number of HVR definitions are used and are included in this document. In some embodiments, the HVRs may be Kabat's complementarity determining regions (CDRs), based on sequence variation and most commonly used (Kabat et al., supra). In some embodiments, the HVRs may be Chothia CDRs. The Chothia system is distinguished by its use of an arrangement of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). In some embodiments, the HVRs may be AbM HVRs. AbM HVRs are a compromise between Kabat CDRs and Chotia structural loops, and are used in Oxford Molecular's AbM antibody modeling program. In some implementations, the HVRs may be contact HVRs. Contact HVRs are based on DOS analysis

- 13 046146 blunt structures of complex crystals. Remains of each of these HVRs are listed below.

Loop Kabat ABM Chotia Contact

L1 L24-L34 L24-L34 L26-L32 L30-L36 L2 L50-L56 L50-L56 L50-L52 L46-L55 L3 L89-L97 L89-L97 L91-L96 L89-L96 H1 Н31-Н35В Н26-Н35В H26-H32 Н30-Н35В (numbering according to Kabat) H1 H31-H35 H26-H35 H26-H32 H30-H35 (numbering according to Chotia) H2 H50-H65 H50-H58 H53-H55 H47-H58 FROM N95-N102 N95-N102 N96-N101 N93-N101

HVRs may include extended HVR as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, and 26 -35 (H1), 50-65 or 49-65 (preferred embodiment) (H2), and 93-102, 94-102, or 95-102 (H3) in VH. The variable domain residues are numbered according to Kabat et al., supra, for each of these extended HVR definitions.

Framework or FR residues are variable domain residues other than HVR residues as defined herein.

A human acceptor framework, as used herein, is a framework comprising an amino acid sequence of a _VL or _VH framework derived from a human immunoglobulin framework or a human consensus framework. A human acceptor scaffold derived from a human immunoglobulin scaffold or a human consensus scaffold may contain the same amino acid sequence thereof, or it may contain predetermined changes to the amino acid sequence. In some embodiments, the number of specified amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In cases where specified amino acid changes are present in VH, preferably such changes are only at three, two or one of the 71H, 73H and 78H positions; for example, the amino acid residues at these positions may be 71A, 73T and/or 78A. In one embodiment, the human VL acceptor framework is identical in sequence to a human immunoglobulin _VL framework sequence or a human consensus framework sequence.

A human consensus framework is a framework that represents the most frequently occurring amino acid residues in selected human immunoglobulin V _L or V _H framework sequences. Typically, the selection of human immunoglobulin V _L or V _H sequences is made from a subset of variable domain sequences. Typically, a subset of sequences is a subset of Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991). Examples include for VL a subgroup that may be a kappa I, kappa II, kappa III, or kappa IV subgroup, as in Kabat et al., supra. Additionally, for VH, the subgroup may be subgroup I, subgroup II or subgroup III, as in Kabat et al., supra.

Amino acid modification at a specific position, for example, the anti-sortilin antibodies of the present invention, refers to the substitution or deletion of a specific residue, or the insertion of at least one amino acid residue adjacent to a specific residue. An insertion near a particular residue means an insertion one to two residues away from it. The insertion may be N-terminal or C-terminal to a specific residue. The preferred amino acid modification herein is a substitution.

An affinity matured antibody, such as the anti-sortilin antibody of the present invention, is an antibody with one or more changes in one or more of its HVRs that result in increased affinity of the antibody for an antigen, compared to a parent antibody that does not have such change(s). ). In one embodiment, the affinity matured antibody has nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are prepared using procedures known in the art. For example, Marks et al., Bio/Technology 10:779-783 (1992) describes affinity maturation by shuffling the VH and VL domains. Non-specific mutagenesis of HVR and/or framework residues is described, for example, in Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al., J. Mol. Biol. 226:889-896 (1992).

As used herein, the term specifically recognizes or specifically binds refers to measurable and reproducible interactions, such as attraction or binding, between a target and an antibody, such as the anti-sortilin antibody herein.

- 14 046146 an invention that allows the presence of a target to be determined in the presence of a heterogeneous population of molecules, including biological molecules. For example, an antibody, such as the anti-sortilin antibody of the present invention, that specifically or preferentially binds to a target or epitope is an antibody that binds that target or epitope with greater affinity, avidity, more readily, and/or for a longer period of time , how it relates to other targets or other epitopes of the target. This definition also implies that, for example, an antibody (or fragment) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. As such, specific binding or preferential binding does not necessarily require (although it may include) exclusive binding. An antibody that specifically binds to a target may have an association constant of at least about 10 ³ M ^-1 or 10 ⁴ M ^-1 , sometimes about 10 ⁵ M ^-1 or 10 ⁶ M ^-1 , in other cases about 10 ⁶ M ^-1 or 10 ⁷ M ^-1 , from about 10 ⁸ M ^-1 to 10 ⁹ M ^-1 , or from about 10 ¹⁰ M ^-1 to 10 ¹¹ M ^-1 or higher. Various immunoassay formats can be used to select antibodies that are specifically immunoreactive for a particular protein. For example, ELISA immunoassays are commonly used to select monoclonal antibodies that are specifically immunoreactive for a protein. See, for example, Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, for immunoassay formats and conditions that can be used to determine specific immunoreactivity.

As used herein, the interaction between the sortilin protein and the second protein includes, without limitation, protein-protein interaction, physical interaction, chemical interaction, binding, covalent binding and ionic binding. As used herein, an antibody inhibits the interaction between two proteins when the antibody disrupts, weakens or completely eliminates the interaction between the two proteins. The antibody of this invention, or a fragment thereof, inhibits the interaction between two proteins when the antibody or fragment thereof binds to one of the two proteins.

An agonist antibody or activating antibody is an antibody, such as the anti-sortilin agonist antibody of the present invention, that induces (eg, enhances) one or more activities or functions of an antigen upon binding of the antibody to the antigen.

A blocking antibody, antagonist antibody, or inhibitory antibody is an antibody, such as the anti-sortilin antibody of the present invention, that inhibits or reduces (e.g., reduces) the binding of an antigen to one or more ligands following binding of the antibody to the antigen, and/or inhibits or reduces (eg, reduces) one or more activities or functions of an antigen after the antibody binds to the antigen. In some embodiments, blocking antibodies, antagonistic antibodies, or inhibitory antibodies substantially or completely inhibit the binding of an antigen to one or more ligands and/or one or more activities or functions of the antigen.

Antibody effector functions refer to those biological activities that can be attributed to the Fc region (native Fc region sequence or Fc region amino acid sequence variant) of an antibody, and vary depending on the isotype of the antibody.

The term Fc region is used herein to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. Although the boundaries of the immunoglobulin heavy chain Fc region may vary, the Fc region of the human IgG heavy chain is generally defined as extending from the amino acid residue at position Cys226, or Pro230, to its carboxyl terminus. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during production or purification of the antibody, or by recombinant modification of the nucleic acid encoding the heavy chain of the antibody. Accordingly, an intact antibody composition may contain antibody populations with K447 residues completely deleted, antibody populations with K447 residues not deleted, and antibody populations with a mixture of antibodies containing and without the K447 residue. Suitable native sequence Fc regions for use in antibodies of this invention include human IgG1, IgG2, IgG3 and IgG4.

The native sequence Fc region contains an amino acid sequence identical to the native amino acid sequence of the Fc region. Native sequence human Fc regions include native sequence human IgG1 Fc region (non-A and A allotypes); Fc region of human IgG2 with native sequence; Fc region of human IgG3 with native sequence; and the native sequence human IgG4 Fc region, as well as natural variants thereof.

The Fc region variant contains an amino acid sequence that differs from the native Fc region sequence due to at least one amino acid modification, preferably one or more amino acid substitutions. Preferably option Fc

- 15 046146 region has at least one amino acid substitution compared to the native sequence of the Fc region or the Fc region of the parent polypeptide, for example from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions, in the native sequence of the Fc region or in the Fc region of the original polypeptide. The Fc region variant herein will preferably have at least about 80% homology to the native Fc region sequence and/or the Fc region of the parent polypeptide, and most preferably at least about 90% homology thereto, more preferably to at least about 95% homology with them.

Fc receptor or FcR describes a receptor that binds to the Fc region of an antibody. A preferred FcR is a human FcR with a native sequence. Further preferred is an FcR that binds an IgG antibody (gamma receptor) and includes receptors of the FcyRI, FcyRII and FcyRIII subclasses, including allelic variants and alternative splice forms of these receptors, FcyRII receptors include FcyRIIA (activating receptor) and FcyRIIB (inhibitory receptor) , which have similar amino acid sequences, differing mainly in their cytoplasmic domains. The activating receptor FcyRIIA contains an immunoreceptor tyrosine-activated motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcyRIIB contains an immunoreceptor tyrosine inhibitory motif (ITIM) in its cytoplasmic domain. The term FcR as used herein covers other FcRs, including those that will be identified in the future. FcRs may also increase the half-life of serum antibodies. As used herein, percentage (%) amino acid sequence identity and homology to a peptide, polypeptide, or antibody sequence refers to the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence, after sequence alignment and introducing gaps if necessary to achieve the maximum percentage of sequence identity, and without taking into account any conservative substitutions when determining sequence identity. Alignment to determine percent amino acid sequence identity can be performed in a variety of ways available to those skilled in the art, for example, using publicly available computer software such as the BLAST, BLAST-2, ALIGN or MEGALIGN™ (DNASTAR) programs. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms known to those skilled in the art necessary to achieve maximum alignment over the full length of the sequences being compared.

An isolated cell is a molecule or a cell that has been identified and separated from at least one contaminant cell with which it is typically associated in its production environment. In some embodiments, the isolated cell is freed of all components associated with the production environment. The isolated cell is in a form that is different from the form or conditions in which it exists in nature. Isolated cells are different from cells that exist naturally in tissues, organs or individuals. In some embodiments, the isolated cell is a host cell of the present invention.

An isolated nucleic acid molecule encoding an antibody, such as the anti-sortilin antibody of the present invention, is a nucleic acid molecule that is identified and separated from at least one contaminating nucleic acid molecule with which it is typically associated in the environment in which it is produced. Preferably, the isolated nucleic acid is not associated with all components associated with the production environment. The isolated nucleic acid molecules encoding the polypeptides and antibodies of this invention are in a form different from the form or conditions in which they exist in nature. The isolated nucleic acid molecules are thus different from the nucleic acid encoding the polypeptides and antibodies of the present invention naturally present in cells.

The term vector, as used herein, shall refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a plasmid, which is a circular double-stranded DNA into which additional DNA segments can be ligated. Another type of vector is a phage vector. Another type of vector is a viral vector, in which additional DNA segments can be ligated into the viral genome. Some vectors are capable of autonomous replication in the host cell into which they are introduced (eg, bacterial vectors having a bacterial origin of replication and mammalian episomal vectors). Other vectors (eg, non-episomal mammalian vectors) can be integrated into the host cell genome upon introduction into the host cell and thus replicate along with the host genome.

- 16 046146

In addition, some vectors are capable of directing the expression of genes to which they are functionally linked. Such vectors are referred to herein as recombinant expression vectors, or simply expression vectors. In general, expression vectors suitable for use in recombinant DNA techniques are often in the form of plasmids. In this specification, plasmid and vector may be used interchangeably, since plasmid is the most widely used form of vector.

Polynucleotide, or nucleic acid, used interchangeably herein, refers to polymers of nucleotides of any length, and includes DNA and RNA. Nucleotides may be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or analogs thereof, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthesis reaction.

A host cell includes an individual cell or cell culture that can be or has been the recipient of the vector(s) for incorporating polynucleotide inserts. Host cells include the progeny of a single host cell, and the progeny will not necessarily be completely identical (in morphology or genomic DNA complement) to the original parent cell due to natural, accidental, or intentional mutation. The host cell includes cells transfected in vivo with the polynucleotide(s) of the present invention.

Carriers as used herein include pharmaceutically acceptable carriers, excipients or stabilizers that are non-toxic to the cell or mammal exposed thereto at the dosages and concentrations used.

The term "about" as used herein refers to the typical error range for the corresponding value, well known to those skilled in the art. Statement about a value or parameter in this document includes (and describes) implementation options that address that value or parameter per se.

As used herein and in the appended claims, the singular form includes references to the plural unless the context clearly indicates otherwise. For example, a reference to an antibody is a reference to one to a plurality of antibodies, such as molar amounts, and includes equivalents thereof known to those skilled in the art, etc.

It should be understood that the aspect and embodiments of the present invention described herein include comprising, consisting of, and essentially consisting of aspects and embodiments.

Review.

This invention relates, in part, to anti-sortilin antibodies that exhibit one or more improved and/or increased functional characteristics compared to the anti-sortilin antibody, S-60, having a heavy chain variable region and a light chain variable region, as described in WO2016164637. Non-limiting improved and/or increased functional properties include, for example, antibodies capable of binding sortilin with higher affinity, reducing cell surface sortilin levels, reducing the half-maximal effective concentration (EC50) for reducing cell surface sortilin levels, improving the maximum reduction of cell surface sortilin levels, increase the extracellular secretion of a sortilin ligand: e.g., progranulin (PGRN), decrease the half-maximal effective concentration (EC50) to block progranulin (PGRN) binding to sortilin, improve the maximum blocking of progranulin (PGRN) binding to sortilin, or any combination thereof. Also provided herein are anti-sortilin antibodies with different Fc variants that exhibit one or more improved and/or increased functional characteristics, including a decrease in the half-maximal effective concentration (EC50) for reducing cell surface sortilin levels, an increase in the maximum reduction in cell surface sortilin levels, an increase extracellular secretion of sortilin ligand: eg, progranulin (PGRN), decreasing the half-maximal effective concentration (EC ₅₀ ) to block progranulin (PGRN) binding to sortilin, and increasing the maximum blocking progranulin (PGRN) binding to sortilin.

In some embodiments, the anti-sortilin antibodies of the present invention have superior activity in reducing cell surface sortilin levels compared to an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, activity is measured as the half-maximal effective concentration (EC ₅₀ ) when cell surface levels of SORT1 are reduced.

The present invention further relates to methods for producing and using anti-sortilin antibodies as described herein; pharmaceutical compositions containing such antibodies; nucleic acids encoding such antibodies; and host cells containing nucleic acids encoding such antibodies.

In some embodiments, the anti-sortilin antibodies of this invention may

- 17 046146 have one or more activities that are caused, at least in part, by the ability of antibodies to reduce levels (eg, cell surface levels) of sortilin by inducing degradation, down regulation, cleavage, receptor desensitization, and/or lysosomal targeting sortilina. In some embodiments, anti-sortilin antibodies exhibit one or more of the following properties: a) have a dissociation constant (K _D ) with human sortilin that is lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region chains corresponding to S-60; b) Reducing cell surface sortilin levels (eg, reducing cell surface sortilin levels on recombinant cells expressing human sortilin in vitro) with a half-maximal effective concentration (EC ₅₀ ) lower than that of the anti-sortilin heavy chain variable region antibody and light chain variable region corresponding to S-60; c) have a dissociation constant (K _D ) with human sortilin, which can range from about 0.560 nM to about 1.63 nM, for example, when determining K _D by fluorescence-activated cell sorting (FACS); d) have a dissociation constant (K _D ) with human sortilin, which can range from about 0.270 nM to about 2.910 nM, for example, when determining K _D by biolayer interferometry; e) reduce cell surface sortilin levels (eg, reduce cell surface sortilin levels on recombinant cells expressing human sortilin in vitro) with a half-maximal effective concentration (EC ₅₀ ), which may range from about 72.58 pM to about 103, 6 pM, for example, when determining EC ₅₀ in vitro by FACS; f) reducing cell surface sortilin levels with a maximum reduction amount that may range from about 51.33% to 88.76%; g) increase extracellular secretion of PGRN more than anti-sortilin antibodies having a heavy chain variable region and a light chain variable region corresponding to S-60; h) block the binding of PGRN to sortilin (eg, block the binding of PGRN to sortilin on recombinant cells expressing human sortilin in vitro) at a half-maximal effective concentration (EC ₅₀ ), which may range from about 325 nM to about 2.27 nM, for example, when determining EC ₅₀ in vitro by FACS; and/or i) increases the maximum block of progranulin (PGRN) binding to sortilin more than anti-sortilin antibodies having a heavy chain variable region and a light chain variable region corresponding to S-60.

Preferably, the anti-sortilin antibodies of the present invention more potently (e.g., with a lower EC50) reduce cell surface sortilin levels (e.g., to about 3.02-fold) compared to a control anti-sortilin antibody (e.g., a control anti-sortilin antibody having a variable a heavy chain region and a light chain variable region corresponding to S-60) (see, for example, Example 2). In some embodiments, the anti-sortilin antibodies of the present invention more strongly reduce the binding of PGRN to sortilin, with a lower EC ₅₀ value (up to about 5.36-fold) compared to a control anti-sortilin antibody (e.g., a control anti-sortilin antibody having a variable region heavy chain and light chain variable region corresponding to S-60) (see, for example, example 3). Additionally, preferably, the anti-sortilin antibodies of the present invention have a higher affinity (e.g., up to about 2.79 times higher affinity) for sortilin (e.g., a lower K _D value as measured by FACS or biolayer interferometry) compared to the control anti-sortilin antibody (e.g., a control anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60 (see, e.g., Examples 1 and 4). Surprisingly, higher affinity for sortilin does not necessarily correlate with either an increase in the ability or activity to reduce cell surface sortilin levels (see, for example, Examples 2 and 4), nor an increase in the ability or activity to block the binding of PGRN to sortilin (see, for example, Examples 3 and 4).

The present invention further provides anti-sortilin antibodies with improved stability during production, storage, and in vivo administration. In some embodiments, the anti-sortilin antibodies of this invention have improved stability under various stress conditions (see, for example, Example 4).

Sortilin proteins.

In one aspect, the present invention provides antibodies, such as isolated (eg, monoclonal) antibodies, that react with, or otherwise bind to, a region, such as an epitope, in the sortilin protein of the present invention. In some embodiments, antibodies react with, or otherwise bind to, a region, such as an epitope, in the sortilin protein of the present invention with improved/increased kinetics (e.g., compared to an anti-sortilin antibody having a heavy chain variable region and a light chain variable region , corresponding to S-60). In some embodiments, the antibodies react with, or otherwise bind to, a region, such as an epitope, in the human sortilin protein, with a half-maximal effective concentration (EC ₅₀ ) that is lower than the value for the control antibody (e.g., compared to the antibody

- 18 046146 against sortilin having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention bind to the sortilin protein and modulate one or more activities of sortilin upon binding to the sortilin protein, for example, an activity associated with the expression of sortilin on a cell. The sortilin proteins of the present invention include, but are not limited to, mammalian sortilin protein, human sortilin protein, mouse sortilin protein, and rat sortilin protein. Typical examples of sortilin protein sequences are given in table. 32.

Sortilin is variously called sortilin 1, SORT1, 100 kDa NT (neurotensin) receptor, glycoprotein 95 (GP95), progranulin receptor (PGRN-R), and neurotensin receptor-3 (NT-3 or NTR-3). Sortilin is an 831 amino acid protein that encodes a type I membrane receptor. Various sortilin homologues are known, including, but not limited to, human sortilin, rat sortilin, and murine sortilin. The amino acid sequence of human sortilin is given below as SEQ ID NO: 81 (key amino acid residues predicted to be involved in progranulin binding are in bold and the predicted pro-NGF (nerve growth factor) binding site is underlined 10 20 30 40 50

MERPWGAADG LSRWPHGLGL LLLLQLLPPS TLSQDRLDAP PPPAAPLPRW

70 80 90 100

SGPIGVSWGL RAAAAGGAFP RGGRWRRSAP GEDEECGRVR DFVAKLANNT

PO 120 130 140 150

HQHVFDDLRG SVSLSWVGDS TGVILVLTTF HVPLVIMTFG QSKLYRSEDY

160 170 180 190 200

GKNFKDITDL INNTFIRTEF GMAIGPENSG KVVLTAEVSG GSRGGRIFRS

210 220 230 240 250

SDFAKNFVQT DLPFHPLTQM MYSPQNSDYL LALSTENGLW VSKNFGGKWE

260 270 280 290 300

EIHKAVCLAK WGSDNTIFFT TYANGSCKAD LGALELWRTS DLGKSFKTIG

310 320 330 340 350

VKIYSFGLGG RFLFASVMAD KDTTRRIHVS TDQGDTWSMA QLPSVGQEQF

360 370 380 390 400

YSILAANDDM VFMHVDEPGD TGFGTIFTSD DRGIVYSKSL DRHLYTTTGG

410 420 430 440 450

ETDFTNVTSL RGVYITSVLS EDNSIQTMIT FDQGGRWTHL RKPENSECDA

460 470 480 490 500

TAKNKNECSL HIHASYSISQ KLNVPMAPLS EPNAVGIVIA HGSVGDAISV

510 520 530 540 550

MVPDVYISDD GGYSWTKMLE GPHYYTILDS GGIIVAIEHS SRPINVIKFS

560 570 580 590 600

- 19 046146

TDEGQCWQTY TFTRDPIYFT GLASEPGARS MNISIWGFTE SFLTSQWVSY

610 620 630 640 650

TIDFKDILER NCEEKDYTIW LAHSTOPEDY EDGCILGYKE QFLRLRKSSV

660 670 680 690 700

CQNGRDYVVT KQPSICLCSL EDFLCDFGYY RPENDSKCVE QPELKGHDLE

710 720 730 740 750

FCLYGREEHL TTNGYRKIPG DKCQGGVNPV REVKDLKKKC TSNFLSPEKQ

760 770 780 790 800

NSKSNSVPII LAIVGLMLVT VVAGVLIVKK YVCGGRFLVH RYSVLQQHAE

810 820 830

ANGVDGVDAL DTASHTNKSG YHDDSDEDLL E

The amino acid sequence of murine sortilin is shown in SEQ ID NO: 82:

MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP

PAPPLLRWAG

PVGVSWGLRA

AAPGGPVPRA

GRWRRGAPAE

HVFDDLSGSV

SLSWVGDSTG

NFKDITNLIN

NTFIRTEFGM

VILVLTTFQV

AIGPENSGKV

DQDCGRLPDF PLVIVSFGQS ILTAEVSGGS

IAKLTNNTHQ

KLYRSEDYGK

RGGRVFRSSD

FAKNFVQTDL

HKAVCLAKWG

PFHPLTQMMY PNNIIFFTTH

SPQNSDYLLA

VNGSCKADLG

LSTENGLWVS

ALELWRTSDL

KNFGEKWEEI

GKTFKTIGVK

IYSFGLGGRF

LFASVMADKD

TTRRIHVSTD

ILAANEDMVF

MHVDEPGDTG

FGTIFTSDDR

QGDTWSMAQL

GIVYSKSLDR

PSVGQEQFYS

HLYTTTGGET

DFTNVTSLRG

VYITSTLSED

KNKNECSLHI

PDVYISDDGG

HASYSISQKL

YSWAKMLEGP

NSIQSMITFD

NVPMAPLSEP

QGGRWEHLRK

NAVGIVIAHG

PENSKCDATA

SVGDAISVMV

HYYTILDSGG

IIVAIEHSNR

PINVIKFSTD

EGQCWQSYVF

DFKDILERNC

TQEPIYFTGL

EEDDYTTWLA

ASEPGARSMN

HSTDPGDYKD

ISIWGFTESF

NGRDYVVAKQ

LYGKEEHLTT

PSVCPCSLED

FLCDFGYFRP

NGYRKIPGDK

KSNSVPIILA IVGLMLVTVV

CQGGMNPARE

AGVLIVKKYV

GCILGYKEQF ENASECVEQP VKDLKKKCTS

CGGRFLVHRY

ITRQWVSYTV LRLRKSSVCQ ELKGHELEFC

NFLNPTKQNS

SVLQQHAEAD

GVEALDSTSH AKSGYHDDSD EDLLE

The amino acid sequence of rat sortilin is shown in SEQ ID NO: 83:

MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG PVGVSWGLRA AAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HVFDDLSGSV SLSWVGDSTG VILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGM AIGPENSGKV ILTAEVSGGS RGGRVFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLA LSTENGLWVS KNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDL GKTFKTIGVK IYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYS ILAANDDMVF MHVDEPGDTG FGTIFTSDDR GIVYSKSLDR HLYTTTGGET DFTNVTSLRG VYITSTLSED NSIQSMITFD QGGRWEHLQK PENSKCDATA KNKNECSLHI HASYSISQKL NVPMAPLSEP NAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMLEGP HYYTILDSGG IIVAIEHSNR PINVIKFSTD EGQCWQSYVF SQEPVYFTGL ASEPGARSMN ISIWGFTESF LTRQWVSYTI DFKDILERNC EENDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQ NGRDYVVAKQ PSICPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTT NGYRKIPGDR CQGGMNPARE VKDLKKKCTS NFLNPKKQNS KSSSVPIILA] CVGLMLVTVV AGVLIVKKYV CGGRFLVHRY SVLQQHAEAD

GVEALDTASH AKSGYHDDSD EDLLE

In some embodiments, sortilin is a preprotein including a signal sequence. In some embodiments, sortilin is a mature protein. In some embodiments, the mature sortilin protein does not include a signal sequence. In some embodiments, the mature sortilin protein is expressed on the cell.

The sortilin proteins of the present invention comprise multiple domains, including, but not limited to, signal sequence, propeptide, luminal domain, Vps10p domain, 10 CC domain, transmembrane domain, and cytoplasmic domain. Additionally, the proteins of this invention are expressed at high levels in a number of tissues, including, without limitation, the brain, spinal cord, cardiac and skeletal muscle, thyroid, placenta and testes.

Accordingly, as used herein, the sortilin protein of this invention includes, without limitation, mammalian sortilin protein, human sortilin protein, primate sortilin protein, murine sortilin protein, and rat sortilin protein. Additionally, the anti-sortilin antibodies of the present invention can bind an epitope of one or more of the sortilin protein

- 20 046146 mammalian thylin, human sortilin protein, primate sortilin protein, mouse sortilin protein, and rat sortilin protein.

Domains of the sortilin protein.

The sortilin proteins of the present invention contain several domains, such as the Vps10p domain, which contains an Asp box motif, a ten-blade beta-propeller structure, and a hydrophobic loop; and domain 10 CC.

As disclosed herein, interactions between the sortilin proteins of the present invention and proneurotrophins or neurotrophins are mediated by the Vps10p domain, which contains a ten-blade beta-propeller structure and an Asp box motif. In some embodiments, the sortilin proteins of the invention comprise a Vps10p domain that includes a ten-blade beta-propeller structure and is located at amino acid residues 78-611 of human sortilin (SEQ ID NO: 81) or amino acid residues of mammalian sortilin that correspond to amino acid residues 78 -611 SEQ ID NO: 81. In some embodiments, amino acid residues 190-220 of human sortilin (SEQ ID NO: 81) or amino acid residues of mammalian sortilin, which correspond to amino acid residues 190-220 of SEQ ID NO: 81, are located in the Vps10p domain .

The Vps10p domains of the present invention may include an Asp box motif. As used herein, Asp box motifs have the following sequence: (S/T)-X-(D/N)-X-XX-X-(W/F/Y) (SEQ ID NO: 84), or X-X-(S/T)-X-(D/N)-X-G-X-(T/S)-(W/F/Y)-X (SEQ ID NO: 85), where X is any amino acid. In human sortilin, the Asp box motif is SSDFAKNF (SEQ ID NO: 86), and is located at amino acid residues 200-207 of human sortilin. Accordingly, in some embodiments, the Asp box motif is located at amino acid residues 200-207 of human sortilin (SEQ ID NO: 81) or amino acid residues of mammalian sortilin that correspond to amino acid residues 200-207 of SEQ ID NO: 81.

As disclosed herein, interactions between the sortilin proteins of this invention and p75 are mediated by the 10CC domain of the hydrophobic loop domain of Vps10p.

In some embodiments, the sortilin proteins of the present invention contain a 10CC domain that is located at amino acid residues 610-757 of human sortilin (SEQ ID NO: 81) or amino acid residues of mammalian sortilin that correspond to amino acid residues 610-757 of SEQ ID NO: 81. In preferred embodiments, amino acid residues 592-593, 610-660, and/or 667-749 of human sortilin (SEQ ID NO: 81) or amino acid residues of mammalian sortilin that correspond to amino acid residues 592-593, 610-660, and/ or 667-749 SEQ ID NO: 81, located in the 10CC domain of sortilin.

In other embodiments, the sortilin proteins of the present invention contain a hydrophobic loop in the Vps10p domain, which is located at amino acid residues 130-141 of human sortilin (SEQ ID NO: 81) or amino acid residues of mammalian sortilin that correspond to amino acid residues 130-141 of SEQ ID NO : 81.

One skilled in the art will appreciate that the starting and ending residues of the domains of this invention may vary depending on the computer modeling program used or the method used to define the domain.

Sortilin binding partners.

The sortilin proteins of this invention may interact with (eg, bind to) one or more proteins, including, without limitation, progranulin protein (PGRN); neurotrophins such as proneurotrophins, proneurotrophin-3, neurotrophin-3, proneurotrophin-4/5, neurotrophin4/5, pro-nerve growth factor (pro-NGF), nerve growth factor (NGF), pro-brain neurotrophic factor (pro- BDNF), and brain-derived neurotrophic factor (BDNF); neurotensin, p75, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), amyloid precursor protein, amyloid beta peptide (A beta), PCSK9, p75NTR, and receptor associated protein (RAP).

Progranulin (PGRN).

The sortilin proteins of the present invention have been demonstrated to interact (eg, bind) directly with progranulin and mediate the degradation of progranulin (eg, Zheng, Y et al., (2011) PLoS ONE 6(6): e21023).

Progranulin is otherwise called PGRN, proepithelin, granulin-epithelin precursor, PC cell growth factor (PCDGF), and acrogranin. Progranulin is a 593 amino acid protein encoding a 68.5 kDa secreted glycoprotein having 7.5 repeats of smaller granulin (epithelin) motifs, ranging in size from 6–25 kDa, which can be proteolytically cleaved from the PGRN precursor. Examples of progranulin cleavage products include, but are not limited to, granulin A/epithelins 1, granulin B/epithelins 2, granulin C, granulins D, granulin E, granulin F, granulin G, and any other known peptide products derived from progranulin.

Accordingly, the anti-sortilin antibodies of the present invention that increase progranulin levels, decrease cell surface sortilin levels, and/or block the interaction (eg, binding) between sortilin and progranulin will be useful in preventing, reducing

- 21 046146 risk management or treatment of conditions and/or diseases associated with reduced levels of progranulin expression and/or activity, cell death (eg, neuronal cell death), frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, traumatic brain damage, spinal cord injury, long-term depression, atherosclerotic vascular disease, undesirable symptoms of normal aging, dementia, mixed dementia, Creutzfeldt-Jakob disease, normal pressure hydrocephalus, amyotrophic lateral sclerosis, Huntington's disease, tauopathy, stroke, acute trauma, chronic trauma, lupus, acute and chronic colitis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, malaria, essential tremor, central nervous system lupus, Behçet's disease, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, intervertebral disc degeneration, Shy-Drager syndrome , progressive supranuclear palsy, corticobasal ganglion degeneration, acute disseminated encephalomyelitis, granulomatous disorders, sarcoidosis, diseases of aging, age-related macular degeneration, glaucoma, retinitis pigmentosa, retinal degeneration, respiratory tract infection, sepsis, eye infection, systemic infection, inflammatory disorders, arthritis, multiple sclerosis, metabolic disorder, obesity, insulin resistance, type 2 diabetes, tissue or vascular damage, trauma, and/or one or more undesirable symptoms of normal aging. Additionally, the anti-sortilin antibodies of the present invention that increase progranulin levels, decrease cell surface sortilin levels, and/or block the interaction (e.g., binding) between sortilin and progranulin may inhibit the interaction between sortilin and progranulin may induce one or more progranulin activities , may reduce the endosomal internalization of progranulin or its fragments, and/or may increase the effective concentration of progranulin.

In some embodiments, the anti-sortilin antibodies of the present invention that increase progranulin levels, decrease cell surface sortilin levels, and/or block the interaction (e.g., binding) between sortilin and progranulin bind to one or more amino acids from amino acid residues 131-138 , 175-181, 190-220, 199-220, 190-211, 196-207, 196-199, 200-207, 203-207, 207-231, 207-227, 212-221, 233-243, 237 -247, 237-260, 297-317, 314338, 367-391, 429-443, 623-632, and/or 740-749 human sortilin (SEQ ID NO: 81); or with amino acid residues of mammalian sortilin, which correspond to amino acid residues 131-138, 175-181, 190-220, 199-220, 190-211, 196-207, 196-199, 200-207, 203-207, 207-231 , 207-227, 212221, 233-243, 237-247, 237-260, 297-317, 314-338, 367-391, 429-443, 623-632, and/or 740-749 SEQ ID NO: 81 In other embodiments, the anti-sortilin antibodies of the present invention, which increase progranulin levels, decrease cell surface sortilin levels, and/or block the interaction (e.g., binding) between sortilin and progranulin, can bind one or more amino acids from amino acid residues His131, Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys 260, Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg382, Tyr386, Ser595, and/or Glu700 of human sortilin (SEQ ID NO: 81); or mammalian sortilin amino acid residues that correspond to one or more amino acid residues His131, Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227 , Ser242, Lys243, Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg382, Ty r386, Ser595, and/or Glu700 SEQ ID NO: 81.

Other sortilin ligands.

The sortilin proteins of the present invention have been shown to interact (eg, bind) directly with proneurotrophins (eg, pro-NGF), which bear a pro-domain and are typically pro-apoptotic. Such binding may be mediated through a linear epitope on sortilin, which corresponds to amino acid residues 163-174 of SEQ ID NO: 81. The sortilin proteins of the present invention have also been shown to interact (eg, bind) to neurotensin in the sortilin beta-propeller structure, and have been an important contact was shown to be at serine at position 283 of human sortilin.

The sortilin proteins of the present invention have also been shown to interact (eg, bind) to the low-affinity nerve growth factor (NGF) receptor (p75) in the sortilin 10CC domain or the hydrophobic loop of the sortilin Vps10p domain. As disclosed herein, the sortilin proteins of the present invention can function as a co-receptor with p75 for proneurotrophins that induce apoptotic signaling. Additionally, the sortilin proteins of the present invention have been shown to interact with (eg, bind to) amyloid precursor protein (APP).

The sortilin proteins of this invention have been shown to interact (e.g., bonds

- 22 046146) with lipoprotein lipase (LpL). As disclosed herein, the sortilin proteins of this invention bind to and modify the degradation of LpL. The sortilin proteins of the present invention have been shown to interact (eg, bind) to apolipoprotein AV (APOA5). As disclosed herein, the sortilin proteins of this invention bind to and modify the degradation of APOA5.

The sortilin proteins of the present invention have been shown to interact with (eg, bind to) apolipoprotein E (APOE, APOE2, APOE3, APOE4). As disclosed herein, the sortilin proteins of this invention bind to and modify the degradation and transport of APOE as well as agents that transfer APOE, such as amyloid beta peptide (A beta). In addition, the sortilin proteins of the present invention have been shown to interact with (eg, bind to) receptor associated protein (RAP). In addition, the sortilin proteins of the present invention have been shown to interact with (eg, bind to) proprotein convertase subtilisin/kexin type 9 (PCSK9), and secrete it into the circulation.

In some embodiments of any of the anti-sortilin antibodies, the anti-sortilin antibodies of the present invention may also inhibit (eg, block) the interaction between sortilin and one or more other sortilin ligands of the present invention. In some embodiments, the one or more other ligands are one or more of proneurotrophins, neurotensin, low-affinity nerve growth factor (NGF) receptor (p75), amyloid precursor protein, lipoprotein lipase, apolipoprotein AV, apolipoprotein, receptor-associated protein, and /or proprotein convertase subtilisin/kexin type 9. Such antibodies may be useful for preventing, reducing the risk of, or treating conditions and/or diseases associated with decreased levels of expression and/or activity of one or more other sortilin ligands, cell death (e.g. neuronal cells). In some embodiments, the one or more other ligands are one or more of proneurotrophins, neurotensin, low-affinity nerve growth factor (NGF) receptor (p75), amyloid precursor protein, lipoprotein lipase, apolipoprotein AV, apolipoprotein, receptor-associated protein, and/or proprotein convertase subtilisin/kexin type 9.

Anti-sortilin antibodies of the present invention that inhibit (eg, block) the interaction between sortilin and one or more other sortilin ligands of the present invention may also prevent cell death (eg, apoptosis) induced by one or more other sortilin ligands. In some embodiments, the one or more other ligands are one or more of proneurotrophins, neurotensin, low-affinity nerve growth factor (NGF) receptor (p75), amyloid precursor protein, lipoprotein lipase, apolipoprotein AV, apolipoprotein, receptor-associated protein, and /or proprotein convertase subtilisin/kexin type 9.

In some embodiments of any of the anti-sortilin antibodies, the anti-sortilin antibodies of the present invention that inhibit (e.g., block) the interaction between sortilin and one or more sortilin ligands of the present invention bind one or more amino acids at amino acid residues 131-138, 175 -181, 190-220, 199-220, 190-211, 196-207, 196-199, 200-207, 203-207, 207-231, 207-227, 212-221, 233-243, 237-247 , 237-260, 297-317, 314-338, 367391, 429-443, 623-632, and/or 740-749 human sortilin (SEQ ID NO: 81); or in amino acid residues of mammalian sortilin, which correspond to amino acid residues 131-138, 175181, 190-220, 199-220, 190-211, 196-207, 196-199, 200-207, 203-207, 207-231, 207 -227, 212-221, 233-243, 237-247, 237-260, 297-317, 314-338, 367-391, 429-443, 623-632, and/or 740-749 SEQ ID NO: 81 In some embodiments, the one or more other ligands are one or more of proneurotrophins, neurotensin, low-affinity nerve growth factor (NGF) receptor (p75), amyloid precursor protein, lipoprotein lipase, apolipoprotein AV, apolipoprotein, receptor-associated protein, and/or proprotein convertase subtilisin/kexin type 9.

In other embodiments of any of the anti-sortilin antibodies, the anti-sortilin antibodies of the present invention that inhibit (e.g., block) the interaction between sortilin and one or more sortilin ligands of the present invention bind one or more amino acids from amino acid residues His131, Val132, Pro133 , Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260, Ser305 , Phe306, Gly307, Arg311 human sortilin (SEQ ID NO: 81); or mammalian sortilin amino acid residues that correspond to one or more amino acid residues His131, Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227 , Ser242, Lys243, Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg382, Ty r386, Ser595, and/or Glu700 SEQ ID NO: 81. In some embodiments, the one or more other ligands are one or more of proneurotrophins, neurotensin, low affinity growth factor receptor ner

- 23 046146 bov (NGF) (p75), amyloid precursor protein, lipoprotein lipase, apolipoprotein AV, apolipoprotein, receptor-associated protein, and/or proprotein convertase subtilisin/kexin type 9.

Sortilin antibodies.

Certain aspects of the present invention relate to anti-sortilin antibodies having one or more improved and/or enhanced functional characteristics. In some embodiments, the anti-sortilin antibodies of the present invention have one or more improved and/or increased functional characteristics compared to the anti-sortilin antibody, S-60, having a heavy chain variable region and a light chain variable region, as described in WO2016164637. In some embodiments, the anti-sortilin antibodies of the present invention have an affinity for sortilin (e.g., human sortilin) that is greater than that of a control anti-sortilin antibody (e.g., a control anti-sortilin antibody containing a heavy chain variable region and a light chain variable region, corresponding to S60). In some embodiments, the anti-sortilin antibodies of the present invention reduce cellular levels (e.g., cell surface levels) of sortilin to a greater extent and with a half-maximal effective concentration (EC50) that is lower than that of a control antibody (e.g., a control anti-sortilin antibody containing a variable heavy chain region and light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention increase the maximum reduction in cell surface sortilin levels compared to an anti-sortilin antibody comprising a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, the anti-sortilin antibodies of the present invention increase the secretion of extracellular progranulin (PGRN) compared to an anti-sortilin antibody comprising a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, the anti-sortilin antibodies of the present invention block the binding of PGRN to sortilin to a greater extent and at a half-maximal effective concentration (EC50) that is lower than that of a control antibody (e.g., a control anti-sortilin antibody containing a heavy chain variable region and a variable region light chain corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention increase the maximum block of progranulin (PGRN) binding to sortilin compared to an anti-sortilin antibody comprising a heavy chain variable region and a light chain variable region corresponding to S-60.

Also provided herein are anti-sortilin antibodies with different Fc variants that exhibit one or more improved and/or increased functional characteristics compared to an anti-sortilin antibody containing a heavy chain variable region and a light chain variable region corresponding to S-60, including a decrease half-maximal effective concentration (EC50) to reduce cell surface sortilin levels, increase maximum reduction of cell surface sortilin levels, increase extracellular secretion of PGRN, decrease half-maximal effective concentration (EC50) to block binding of progranulin (PGRN) to sortilin, and increase maximum blocking of progranulin binding ( PGRN) with sortilin.

In some embodiments, the anti-sortilin antibody of the invention is a human antibody, a bispecific antibody, a monoclonal antibody, a multivalent antibody, a conjugate antibody, or a chimeric antibody.

In a preferred embodiment, the anti-sortilin antibody of this invention is a monoclonal antibody.

Epitope-binding antibody against sortilin.

In some embodiments, the anti-sortilin antibodies of the present invention bind to the sortilin protein of the present invention and/or natural variants. In some embodiments, the anti-sortilin antibodies of the present invention bind to the sortilin protein, wherein the sortilin protein is a human protein. In some embodiments, the anti-sortilin antibodies of the present invention bind to the sortilin protein, wherein the sortilin protein is a wild-type protein. In some embodiments, the anti-sortilin antibodies of the present invention bind to the sortilin protein, wherein the sortilin protein is a naturally occurring variant.

In certain preferred embodiments, the anti-sortilin antibodies of this invention specifically bind to the human sortilin protein.

Certain aspects of the present invention provide anti-sortilin antibodies that bind a discontinuous epitope of sortilin. In some embodiments, the discontinuous sortilin epitope comprises two or more peptides, three or more peptides, four or more peptides, five or more peptides, six or more peptides, seven or more peptides, eight or more peptides, nine or more peptides, or 10 or more peptides. In some implementations

- 24 046146 tions, each of the peptides contains five or more, six or more, seven or more, eight or more, nine or more, 10 or more, 11 or more, 12 or more, 13 or more 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, 24 or more, 25 or more, 26 or more, 27 or more, 28 or more, 29 or more, or 30 or more amino acid residues of the amino acid sequence SEQ ID NO: 81; or five or more, six or more, seven or more, eight or more, nine or more, 10 or more, 11 or more, 12 or more, 13 or more 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, 24 or more, 25 or more, 26 or more, 27 or more, 28 or more, 29 or more, or 30 or more amino acid residues on a mammalian sortilin protein corresponding to the amino acid sequence of SEQ ID NO: 81. Other aspects of the invention provide anti-sortilin antibodies that bind to a conformational epitope of sortilin.

Certain aspects of the present invention provide anti-sortilin antibodies that bind to one or more amino acids from amino acid residues 207-231 of human sortilin (SEQ ID NO: 81); or to amino acid residues of mammalian sortilin corresponding to amino acid residues 207-231 of SEQ ID NO: 81. In some embodiments, anti-sortilin antibodies bind to one or more amino acid residues of amino acid residues Thr218, Tyr222, Ser223, and/or Ser227 of human sortilin (SEQ ID NO: 81); or mammalian sortilin amino acid residues corresponding to amino acid residues Thr218, Tyr222, Ser223, and/or Ser227 of SEQ ID NO: 81. In some embodiments, anti-sortilin antibodies bind to one or more amino acid residues of amino acid residues F105, L108, R109, G110, I537 , F569, E590, F592, L593, S595, and/or W597 human sortilin (SEQ ID NO: 81); or mammalian sortilin amino acid residues corresponding to amino acid residues F105, L108, R109, G110, I537, F569, E590, F592, L593, S595, and/or W597 SEQ ID NO: 81. Other aspects of the invention provide anti-sortilin antibodies that bind with an epitope containing the amino acid residues (S/T)-X-(D/N)-X-X-X-X-(W/F/Y), where X is any amino acid (SEQ ID NO: 84).

In some embodiments, the anti-sortilin antibodies of the invention bind to the sortilin protein of the invention expressed on the cell surface, and the naked antibodies inhibit the interaction (e.g., binding) between the sortilin protein and a protein selected from progranulin (PGRN), proneurotrophin, neurotrophin , proneurotrophin-3, neurotrophin-3, proneurotrophin-4/5, neurotrophin-4/5, pro-nerve growth factor (pro-NGF), nerve growth factor (NGF), pro-brain-derived neurotrophic factor (pro-BDNF), brain-derived neurotrophic factor (BDNF), neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E ( APOE), PCSK9, and receptor-associated protein (RAP). In some embodiments, the anti-sortilin antibodies of the invention bind to the sortilin protein of the invention expressed on the cell surface, and the naked antibodies inhibit the interaction (eg, binding) between the sortilin protein and PGRN.

In some embodiments, anti-sortilin antibodies of the invention that bind to the sortilin protein of the invention inhibit the interaction (e.g., binding) between the sortilin protein and progranulin, proneurotrophin, neurotrophin, proneurotrophin-3, neurotrophin-3, proneurotrophin-4/ 5, neurotrophin-4/5, pro-nerve growth factor (pro-NGF), nerve growth factor (NGF), pro-brain-derived neurotrophic factor (pro-BDNF), brain-derived neurotrophic factor (BDNF), neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and receptor associated protein (RAP) by reducing effective levels of sortilin available to interact with these proteins on the cell surface or within the cell.

In some embodiments, anti-sortilin antibodies of the invention that bind to the sortilin protein of the invention inhibit the interaction (e.g., binding) between the sortilin protein and progranulin, proneurotrophin, neurotrophin, proneurotrophin-3, neurotrophin-3, proneurotrophin-4/ 5, neurotrophin-4/5, pro-nerve growth factor (pro-NGF), nerve growth factor (NGF), pro-brain-derived neurotrophic factor (pro-BDNF), brain-derived neurotrophic factor (BDNF), neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and receptor associated protein (RAP) by inducing degradation of sortilin.

In some embodiments, the anti-sortilin antibodies of the present invention bind to human sortilin or a homolog thereof, including, without limitation, mammalian sortilin protein, or non-human primate sortilin protein. In some implementations,

- 25 046146 anti-sortilin antibodies of this invention specifically bind to human sortilin. In some embodiments, the anti-sortilin antibodies of the present invention bind to human sortilin and are not cross-reactive with sortilin orthologs or homologues from other species.

Competitive binding of anti-sortilin antibody.

In some embodiments, the anti-sortilin antibodies of this invention competitively inhibit the binding of at least one other anti-sortilin antibody selected from any of the antibodies listed in table. 1-29. In some embodiments, the anti-sortilin antibodies of this invention competitively inhibit the binding of at least one antibody selected from: S-60, S-60-1, S-60-2, S-60-3, S-60-4 , S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt) (wild type)], S-6015.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S60 -15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F] , S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60- 16; S-60-18, S-60-19, S-60-24; and any combination thereof.

In some embodiments, the anti-sortilin antibody of the present invention competes with one or more anti-sortilin antibodies selected from: S-60, S-60-1, S-60-2, S-603, S-60-4, S -60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K ], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60 -15.13 [N33I], S-60-15.14 [N33V], S-6015.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24 and any combination thereof, for binding to sortilin when an anti-sortilin antibody reduces the binding of one or more antibodies selected from: S-60, S-60-1 , S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 ( described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T ], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60 -15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S60-15.12 [N33F], S -60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24; and any combination thereof, with sortilin by an amount of from about 50% to 100% compared with binding to sortilin in the absence of anti-sortilin antibody.

In some embodiments, the anti-sortilin antibody of the present invention competes with one or more anti-sortilin antibodies selected from: S-60, S-60-1, S-60-2, S-603, S-60-4, S -60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K ], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60 -15.13 [N33I], S-60-15.14 [N33V], S-6015.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof, for binding to sortilin, when the anti-sortilin antibody reduces the binding of one or more antibodies selected from: S-60, S-60- 1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T ], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60 -15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S60-15.12 [N33F], S -60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24; and any combination thereof, with sortilin at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80% , at least 85%, at least 90%, at least 95%, or 100%, compared to binding to sortilin in the absence of anti-sortilin antibody.

In some embodiments, an anti-sortilin antibody of the invention that reduces the binding of one or more antibodies selected from: S-60, S-60-1, S-60-2, S-60-3, S-60-4 , S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-6015.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60 -15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof, with sortilin 100% indicates that the anti-sortilin antibody substantially completely blocks the binding of one or more antibodies selected from S- 60, S-60-1, S-60-2, S-60-3, S-60-4, S60-5, S-60-6, S-60-7, S-60-8, and S -60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-6013, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S -60-15.13 [N331], S-60-15.14 [N33V], S-60-15.15 [N33A],

- 26 046146

S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof, with sortilin. In some embodiments, an anti-sortilin antibody and one or more antibodies selected from S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T ], S60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S60 -15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S60-18, S-60-19, S-60-24, and any combination thereof, are present in amounts that correspond to a 10:1 ratio, a 9:1 ratio, an 8:1 ratio, a 7:1 ratio, a 6 ratio: 1, 5:1 ratio, 4:1 ratio, 3:1 ratio, 2:1 ratio, 1:1 ratio, 0.75:1 ratio, 0.5:1 ratio, 0.25:1 ratio, 0 ratio .1:1, 0.075:1 ratio, 0.050:1 ratio, 0.025:1 ratio, 0.01:1 ratio, 0.0075:1 ratio, 0.0050:1 ratio, 0 ratio .0025:1, ratio 0.001:1 (0.001: ratio), 0.00075:1, ratio 0.00050:1, ratio 0.00025:1, ratio 0.0001:1 (0.0001: ratio), ratio 1:10, 1:9 ratio, 1:8 ratio, 1:7 ratio, 1:6 ratio, 1:5 ratio, 1:4 ratio, 1:3 ratio, 1:2 ratio, 1:0.75 ratio , ratio 1:0.5, ratio 1:0.25, ratio 1:0.1, ratio 1:0.075, ratio 1:0.050, ratio 1:0.025, ratio 1:0.01, ratio 1:0.0075 , ratio 1:0.0050, ratio 1:0.0025, ratio 1:0.001, ratio 1:0.00075, ratio 1:0.00050, ratio 1:0.00025, or ratio 1:0.0001 antibodies against sortilin to one or more antibodies selected from S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S- 60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-6015.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K ], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60 -15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-6015.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof. In some embodiments, the anti-sortilin antibody is present in a quantitative excess ranging from about 1.5-fold to 100-fold, or greater than 100-fold, over the amount of one or more antibodies selected from S-60, S- 60-1, S-60-2, S-60-3, S-60-4, S-60-5, S60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-6014, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S60-15.11 [N33W], S-60-15.12 [N33F], S-60 -15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof. In some embodiments, the anti-sortilin antibody is present in an amount that is about 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold. multiple, 20x, 25x, 30x, 35x, 40x, 45x, 50x, 55x, 60x, 65x, 70x, 75x, 80-fold, 85-fold, 90-fold, 95-fold, or 100-fold excess over the amount of one or more antibodies selected from S-60, S-60-1, S-60-2, S- 60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T ], S60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S60 -15.13 [N331], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S60-18, S-60-19, S-60-24, and any combination thereof.

In some embodiments, the anti-sortilin antibodies of the present invention bind to a human sortilin epitope that is the same as or overlaps with a sortilin epitope bound by at least one antibody selected from any of the antibodies listed in Table 1. 1-29. In some embodiments, the anti-sortilin antibodies of the present invention bind to an epitope of human sortilin that is the same as or overlaps with an epitope of sortilin bound by at least one antibody selected from: S-60, S-60-1, S-60 -2, S60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T ], S-60-15.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S -60-15.13 [N33I], S-60-15.14 [N33V], S-6015.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24; and any combination thereof. In some embodiments, the anti-sortilin antibodies of this invention bind substantially the same sortilin epitope as that bound by at least one antibody selected from any of the antibodies listed in Table 1. 1-29. In some embodiments, the anti-sortilin antibodies of the present invention bind substantially the same sortilin epitope as that bound by at least one antibody selected from: S-60, S-60-1, S-60-2, S-60 -3, S-60-4, S-60-5, S60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-6014, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5

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[N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S60 -15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M] , S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24; and any combination thereof. A detailed description of typical methods for mapping the epitope to which an antibody binds is given in Morris (1996) Epitope Mapping Protocols, in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ).

Any suitable competition or sortilin binding assay known in the art, such as BIAcore assay, ELISA assays, or flow cytometry, can be used to determine whether an anti-sortilin antibody competes with one or more antibodies selected from S-60, S- 60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60 -9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T ], S60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S60 -15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S60-18, S-60-19, S-60-24; and any combination thereof, for binding to sortilin. In a typical example of a competition assay, immobilized sortilin or cells expressing sortilin on the cell surface are incubated in a solution containing a first labeled antibody that binds to sortilin (e.g., human or non-human primate) and a second unlabeled antibody that is tested for its ability to compete with the first antibody for binding to sortilin. The second antibody may be present in the supernatant of the hybridoma. As a control, immobilized sortilin or cells expressing sortilin are incubated in a solution containing the first labeled antibody but not the second unlabeled antibody. After incubation under conditions permissive for binding of the first antibody to sortilin, excess unbound antibody is removed and the amount of label bound to immobilized sortilin or sortilin-expressing cells is measured. If the amount of label bound to immobilized sortilin or cells expressing sortilin is significantly reduced in the test sample compared to the control sample, then this indicates that the second antibody is competing with the first antibody for binding to sortilin. See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch. 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

In some embodiments, the anti-sortilin antibody of the present invention competes for binding to sortilin with an antibody comprising a heavy chain variable domain and a light chain variable domain of an antibody selected from the group consisting of S-60-10, S-60-11, S -60-12, S60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-16, S-60-18, S- 60-19, S-60-24, and any combination thereof.

In some embodiments, the anti-sortilin antibody of the present invention binds substantially the same sortilin epitope as an antibody comprising a heavy chain variable domain and a light chain variable domain of an antibody selected from the group consisting of S-60-10, S-60 -11, S-6012, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-16, S-60- 18, S-60-19, and S-60-24.

Additional anti-sortilin antibodies, for example, antibodies that specifically bind to the sortilin protein of the present invention, can be identified, screened, and/or characterized for their physical/chemical properties and/or biological activities by various assays known in the art.

Variable regions of the heavy chain and light chain of the anti-sortilin antibody.

A. Heavy chain HVR.

In some embodiments, the anti-sortilin antibodies of the present invention include a heavy chain variable region comprising one or more (e.g., one or more, two or more, or all three) HVRs selected from HVR-H1, HVR-H2, and HVR- H3 (as indicated in Tables 14-16). In some embodiments, the heavy chain variable region comprises HVR-H1, HVR-H2, and HVR-H3 (as set forth in Tables 14-16).

In some embodiments, HVR-H1 contains the sequence YSISSGYYWG (SEQ ID NO: 1). In some embodiments, HVR-H2 contains a sequence in accordance with Formula I: TIYHSGSTYYNPSLX1S (SEQ ID NO: 4), where X1 is K or E. In some embodiments, HVR-H2 contains a sequence selected from SEQ ID NO: 2-3. In some embodiments, HVR-H3 comprises the sequence of Formula II: ARQGSIX1QGYYGMDV (SEQ ID NO: 7). In some embodiments, HVR-H3 comprises a sequence selected from SEQ ID NO: 5-6.

In some embodiments, HVR-H1 comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to the amino acid sequence of SEQ ID

NO: 1. In some embodiments, HVR-H1 contains an amino acid sequence containing substitutions (e.g., conservative substitutions, insertions, or deletions relative to the amino acid sequence of SEQ ID NO: 1), but retains the ability to bind sortilin . In some embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of HVR-H1 SEQ ID NO: 1. In some embodiments, HVR-H2 comprises amino acid sequence with at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96 %, at least about 97%, at least about 98%, at least about 99%, or 100% identity with the amino acid sequence selected from SEQ ID NO: 2-3. In some embodiments, HVR-H2 contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions or deletions compared to the amino acid sequence selected from SEQ ID NO: 2-3), but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acids have been substituted, inserted, and/or deleted in the HVR-H2 amino acid sequence selected from SEQ ID NO: 2-3. In some embodiments, HVR-H3 comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with an amino acid sequence selected from SEQ ID NO: 5-6. In some embodiments, HVR-H3 contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions or deletions compared to the amino acid sequence selected from SEQ ID NO: 5-6), but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acids have been substituted, inserted, and/or deleted in the HVR-H3 amino acid sequence selected from SEQ ID NO: 5-6.

In some embodiments, the heavy chain variable region comprises HVR-H1 containing the sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the sequence in accordance with Formula I, and HVR-H3 containing the sequence in accordance with Formula II.

In some embodiments, the heavy chain variable region comprises HVR-H1 comprising the sequence of SEQ ID NO: 1, HVR-H2 containing the sequence selected from SEQ ID NO: 2-3, and HVR-H3 containing the sequence selected from SEQ ID NO: 5-6.

In some embodiments, the heavy chain variable region comprises HVR-H1, HVRH2, and HVR-H3 antibodies S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S- 60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E ], S-60-15.11 [N33W], S-60-15.12 [N33F], S60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S60-18, S-60-19, S-60-24, or any combination thereof (as indicated in Table 14-16).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region, wherein the heavy chain variable region comprises one or more of: (a) HVR-H1 comprising an amino acid sequence with at least 85% of at least 86 %, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-H1 antibodies S-60-10, S-60 -11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S] , S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60- 15.8 [N33Q], S-6015.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S- 60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; (b) HVR-H2 containing an amino acid sequence of at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% , or 100% identity with the amino acid sequence of HVR-H2 antibodies S-60-10, S-60-11, S60-12, S-60-13, S-60-14, S-60-15 [N33 (wt) ], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S -60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-6015.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L] , S-60-16; S-60-18, S-60-19, or S-60-24; and (c) HVRH3 comprising an amino acid sequence of at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-H3 antibodies S-60-10, S-60-11, S-60-12, S-6013, S-60-14, S-60-15 [N33 (wt)] , S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R],

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S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S -60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

In some embodiments, the anti-sortilin antibodies of the present invention comprise HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVR-H3 containing amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6).

B. Light chain HVR.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising one or more (e.g., one or more, two or more, or all three) HVRs selected from HVR-L1, HVR-L2, and HVR- L3 (as indicated in Table 17-19). In some embodiments, the light chain variable region comprises HVR-L1, HVR-L2, and HVRL3 (as set forth in Tables 17-19).

In some embodiments, HVR-L1 contains the sequence in accordance with Formula III: RSSQX ₁ LLX ₂ SX ₃ GYNYLD (SEQ ID NO: 28), where X1 is S or G, X2 is R or H, and X ₃ is is N, T, S, G, R, D, H, K, Q, Y, E, W, F, I, V, A, M, or L. In some embodiments, HVR-L1 contains the sequence selected from SEQ ID NO: 8-27. In some embodiments, HVR-L1 comprises the sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), RSSQSLLRSTGYNYLD (SEQ ID NO: 9), RSSQS LLRSSGYNYLD (SEQ ID NO: 10), RSSQSLLRSGGYNYLD (SEQ ID NO: 11), RSSQSLLRSRG YNYLD ( SEQ ID NO: 12),

RSSQSLLRSDGYNYLD (SEQ ID NO: 13), RSSQSLLRSHGYNYLD (SEQ ID NO: 14),

RSSQSLLRSKGYNYLD (SEQ ID NO: 15), RSSQSLLRSQGYNYLD (SEQ ID NO: 16), RSSQSLLRSYGYNYLD (SEQ ID NO: 17), RSSQSLLRSEGYNYLD (SEQ ID NO: 18), RSSQSLLRSWGYNYLD (SEQ ID NO: 19), RSSQSLLRSFGYNYLD (SEQ ID NO : 20), RSSQSL LRSIGYNYLD (SEQ ID NO: 21), RSSQSLLRSVGYNYLD (SEQ ID NO: 22), RSSQSLLRSAG YNYLD (SEQ ID NO: 23), RSSQSLLRSMGYNYLD (SEQ ID NO: 24), RSSQSLLRSLGYNYLD (SEQ ID NO: 25) , RSSQSLLHSNGYNYLD (SEQ ID NO: 26), or RSSQGLLRSNGYNYLD (SEQ ID NO: 27). In one particular embodiment, HVR-L1 contains the sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8). In another specific embodiment, HVR-L1 contains the sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9) (as set forth in Table 19).

In some embodiments, HVR-L2 contains a sequence according to Formula IV: LGSNRX1S (SEQ ID NO: 31), where X1 is A or V. In some embodiments, HVR-L2 contains a sequence selected from SEQ ID NO: 29-30.

In some embodiments, HVR-L3 contains a sequence in accordance with Formula V: MQQQEX1PLT (SEQ ID NO: 34), where X1 is A or T. In some embodiments, HVR-L3 contains a sequence selected from SEQ ID NO: 32-33.

In some embodiments, HVR-L1 comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with an amino acid sequence selected from SEQ ID NO: 8-27. In some embodiments, HVR-L1 contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions or deletions compared to the amino acid sequence selected from SEQ ID NO: 8-27), but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acids have been substituted, inserted, and/or deleted in the HVR-L1 amino acid sequence selected from SEQ ID NOs: 8-27. In some embodiments, HVR-L2 comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with the amino acid sequence selected from SEQ ID NO: 29-30. In some embodiments, HVR-L2 contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions, or deletions compared to the amino acid sequence selected from SEQ ID NO: 2930), but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acids have been substituted, inserted, and/or deleted in the HVR-L2 amino acid sequence selected from SEQ ID NOs: 29-30. In some embodiments, HVR-L3 comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with the amino acid sequence selected from SEQ ID NO: 32-33. In some implementations

- 30 046146 tion, HVR-L3 contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions or deletions compared to the amino acid sequence selected from SEQ ID NO: 32-33), but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acids have been substituted, inserted, and/or deleted in the HVR-L3 amino acid sequence selected from SEQ ID NOs: 32-33.

In some embodiments, the light chain variable region comprises HVR-L1 comprising a sequence according to Formula III, HVR-L2 comprising a sequence according to Formula IV, and HVR-L3 comprising a sequence according to Formula V. In some embodiments implementation, the light chain variable region comprises HVR-L1 containing a sequence selected from SEQ ID NO: 8-27, HVR-L2 containing a sequence selected from SEQ ID NO: 29-30, and HVR-L3 containing a sequence selected from from SEQ ID NO: 32-33.

In some embodiments, the light chain variable region comprises HVR-L1, HVR-L2, and HVR-L3 antibodies S-60-10, S-60-11, S-60-12, S-60-13, S-60- 14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S- 60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E ], S-60-15.11 [N33W], S-60-15.12 [N33F], S60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S60-18, S-60-19, S-60-24, or any combination thereof (as indicated in Table 17-19).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region, wherein the light chain variable region comprises one or more of: (a) HVR-L1 comprising an amino acid sequence with at least 85% of at least 86 %, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-L1 antibodies S-60-10, S60-11 , S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S -60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [ N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S- 60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; (b) HVR-L2 containing an amino acid sequence of at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% , or 100% identity with the amino acid sequence of HVR-L2 antibodies S-60-10, S-60-11, S60-12, S-60-13, S-60-14, S-60-15 [N33 (wt) ], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S -60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-6015.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L] , S-60-16; S-60-18, S-60-19, or S-60-24; and (c) HVRL3 comprising an amino acid sequence of at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-L3 antibodies S-60-10, S-60-11, S-60-12, S-6013, S-60-14, S-60-15 [N33 (wt)] , S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60- 15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

In some embodiments, the anti-sortilin antibodies of the present invention comprise HVR-L1 comprising the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 comprising the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 comprising amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, the anti-sortilin antibodies of the present invention comprise HVR-L1 comprising the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 comprising the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 comprising amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

C. Heavy chain HVR and light chain HVR.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising one or more (e.g., one or more, two or more, or all three) HVRs selected from HVR-H1, HVR-H2, and HVR- H3 (as defined in Tables 14-16), and a light chain variable region comprising one or more (e.g., one or more, two or more, or all three) HVRs selected from HVR-L1, HVR-L2, and HVR -L3 (as indicated in Tables 17-19). In some embodiments, the heavy chain variable region comprises HVR-H1, HVR-H2, and

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HVR-H3 (as listed in Tables 14-16), and the light chain variable region contains HVR-L1, HVR-L2 and HVR-L3 (as listed in Tables 17-19).

In some embodiments, the heavy chain variable region comprises HVR-H1 containing the sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the sequence in accordance with Formula I, and HVR-H3 containing the sequence in accordance with Formula II, and the light chain variable region comprises HVR-L1 comprising a sequence according to Formula III, HVR-L2 comprising a sequence according to Formula IV, and HVR-L3 comprising a sequence according to Formula V. In some embodiments, the variable region heavy chain contains HVR-H1 containing the sequence selected from SEQ ID NO: 1, HVRH2 containing the sequence selected from SEQ ID NO: 2-3, and HVR-H3 containing the sequence selected from SEQ ID NO: 5-6, and variable the light chain region comprises HVR-L1 containing a sequence selected from SEQ ID NOs: 8-27, HVR-L2 containing a sequence selected from SEQ ID NOs: 29-30, and HVR-L3 containing a sequence selected from SEQ ID NO: 32-33.

In some aspects, the heavy chain variable region comprises HVR-H1 comprising a sequence selected from SEQ ID NO: 1, HVR-H2 comprising a sequence selected from SEQ ID NOs: 2-3, and HVR-H3 comprising a sequence selected from SEQ ID NO: 2-3. NO: 5-6, and the light chain variable region comprises HVR-L1 containing a sequence selected from SEQ ID NOs: 8-27, HVRL2 containing a sequence selected from SEQ ID NOs: 29-30, and HVR-L3 containing sequence SEQ ID NO: 32.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising HVR-H1, HVR-H2 and HVR-H3 antibodies S-60-10, S-6011, S-60-12, S-60- 13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S -60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [ N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, or any combination thereof (as indicated in Table 14-16); and a light chain variable region containing HVR-L1, HVR-L2 and HVR-L3 antibodies S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S -60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60 -15.5 [N33D], S-60-15.6 [N33H], S-6015.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S -60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [ N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, or any combination thereof (as indicated in Table 17-19).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising HVR-H1, HVR-H2 and HVR-H3 and a light chain variable region comprising HVR-L1, HVR-L2 and HVR-L3, with This antibody contains HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 antibodies S-60-10, S-60-11, S-60-12, S-60-13 , S-60-14, S60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [ N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S- 60-15.10 [N33E], S60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as indicated in Table 14-19).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises one or more of: (a) HVR-H1 comprising an amino acid sequence of at least 90% , at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, according to at least 99%, or 100% identity with the amino acid sequence of HVR-H1 antibodies S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60- 15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [ N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S- 60-15.11 [N33W], S-60-15.12 [N33F], S60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S60-18, S-60-19, or S-60-24; (b) HVR-H2 containing an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-H2 antibodies S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-6015.3 [N33G], S -60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [ N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S- 60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; and (c) HVR-H3 comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-H3 antibodies S-60-10, S-60-11, S-60-12 , S-60

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13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60 -15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y] , S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60- 15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; and the light chain variable region comprises one or more of: (a) HVR-L1 comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVRL1 antibodies S-60-10, S-60 -11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S] , S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-6015.8 [ N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S- 60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S60-19, or S-60-24; (b) HVR-L2 containing an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-L2 antibodies S-60-10, S60-11, S-60-12, S- 60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S -60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [ N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S- 60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; and (c) HVR-L3 comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of HVR-L3 antibodies S-60-10, S-60-11, S-60-12 , S-6013, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S -60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRVS (SEQ ID NO: 30), and HVR-L3 containing the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLES (SEQ ID NO: 3), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR -H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region containing HVR-H1 containing the amino acid sequence

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YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLHSNGYNYLD (SEQ ID NO: 26), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), HVR-H3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQGLLRSNGYNYLD (SEQ ID NO: 27), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

D. Heavy chain variable region.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NOs: 54-56. In some embodiments, the heavy chain variable region comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with an amino acid sequence selected from SEQ ID NO: 54-56 . In some embodiments, the heavy chain variable region contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions, or deletions compared to the amino acid sequence selected from SEQ ID NOs: 54-56) but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, up to 5, up to 6, up to 7, up to 8, up to 9, or up to 10 amino acids have been substituted, inserted, and/or deleted in the amino acid heavy chain variable region sequence selected from SEQ ID NO: 54-56.

In some embodiments, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 56.

In some embodiments, the anti-sortilin antibodies of the present invention comprise the antibody heavy chain variable region S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60 -15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S -60-15.11 [N33W], S60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M] , S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as indicated in Table 30).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVR-H2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVR-H3, containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6).

E. Light chain variable region.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising an amino acid sequence selected from SEQ ID NOs: 57-80. In some embodiments, the light chain variable region comprises amino

- 34 046146 acid sequence with at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with an amino acid sequence selected from SEQ ID NOs: 57-80. In some embodiments, the light chain variable region contains an amino acid sequence containing substitutions (eg, conservative substitutions, insertions, or deletions compared to the amino acid sequence selected from SEQ ID NOs: 57-80) but retains the ability to bind sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, up to 5, up to 6, up to 7, up to 8, up to 9, or up to 10 amino acids have been substituted, inserted, and/or deleted in the amino acid light chain variable region sequence selected from SEQ ID NO: 57-80.

In some embodiments, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 57. In some embodiments, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 60.

In some embodiments, the anti-sortilin antibodies of the present invention comprise the antibody light chain variable region S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60 -15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S -60-15.11 [N33W], S60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M] , S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as indicated in Table 31).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3, containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising HVR-L1 comprising the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3, containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

F. Heavy chain variable region and light chain variable region.

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-56; and/or a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 57-80. In some embodiments, the heavy chain variable region comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with an amino acid sequence selected from SEQ ID NO: 54-56 and the light chain variable region comprises an amino acid sequence of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95 %, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity with an amino acid sequence selected from SEQ ID NO: 57-80. In some embodiments, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising an amino acid sequence containing substitutions (e.g., conservative substitutions, insertions, or deletions compared to the amino acid sequence selected from SEQ ID NOs: 54-56), and a light chain variable region containing an amino acid sequence containing substitutions (eg, conservative substitutions, insertions or deletions compared to the amino acid sequence selected from SEQ ID NO: 57-80), but retains the ability to bind to sortilin. In some embodiments, up to 1, up to 2, up to 3, up to 4, up to 5, up to 6, up to 7, up to 8, up to 9, or up to 10 amino acids have been substituted, inserted, and/or deleted in the amino acid heavy chain variable region sequence selected from SEQ ID NO: 54-56; and up to 1, up to 2, up to 3, up to 4, up to 5, up to 6, up to 7, up to 8, up to 9, or up to 10 amino acids have been substituted, inserted and/or deleted in the light chain variable region of an amino acid sequence selected from SEQ ID NO: 57-80.

In some aspects, the anti-sortilin antibody of the present invention comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-56; and/or a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 57-58, 60-78, and 80.

In some embodiments, the anti-sortilin antibody of this invention binds

- 35 046146 combines with the protein sortilin, wherein the antibody contains a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 54, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 57; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 54, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 58; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 54, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 59; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 55, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 57; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 55, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 58; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 57; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 77; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 78; a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 54, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 79; or a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 80.

In one aspect, the anti-sortilin antibody of the present invention comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 56 and a light chain variable region having the amino acid sequence of SEQ ID NO: 57.

In one aspect, the anti-sortilin antibody of the present invention comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 56 and a light chain variable region having the amino acid sequence of SEQ ID NO: 60.

In some embodiments, the anti-sortilin antibodies of the present invention comprise the antibody heavy chain variable region S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60 -15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S -60-15.11 [N33W], S60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M] , S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as specified in Table 30), and the antibody light chain variable region S-60-10, S-60-11, S-60-12 , S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G ], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-6015.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S -60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S60-19, or S-60-24 (as indicated in Table 31).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising an amino acid sequence selected from SEQ ID NO: 56 and 60. In some embodiments , the antibody contains the variable region of the heavy chain of S-60-15 [N33 (wt)] (as indicated in Table 30), and the variable region of the light chain of the antibody S-60-15 [N33 (wt)] (as indicated in Table 31). In some embodiments, the antibody comprises an S-60-15.1 [N33T] heavy chain variable region (as set forth in Table 30), and an S-60-15.1 [N33T] antibody light chain variable region (as set forth in Table 31).

Typical examples of antibodies against sortilin.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody comprising a heavy chain variable region and a light chain variable region of an antibody selected from S-60-10, S-60-11, S-60-12, S-60 -13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S- 60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y ], S-60-15.10 [N33E], S-60-15.11 [N33W], S60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19 or S-60-24. In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody comprising a heavy chain and a light chain of an antibody selected from S-60-10, S-60-11, S-60-12, S-60-13, S -60-14, S-60-15 [N33 (wt)], S60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R] , S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60- 15.10 [N33E], S-60-15.11 [N33W], S60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S- 60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

(1) S-60-10.

In some embodiments, the anti-sortilin antibodies of this invention comprise

- 36 046146 a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of the antibody S60-10 or with amino acid sequence SEQ ID NO: 54; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-10 or with the amino acid sequence of SEQ ID NO: 57. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of the antibody S-60-10 or with the amino acid sequence SEQ ID NO: 54, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-10. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-10 or with amino acid sequence of SEQ ID NO: 57, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-10. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of the antibody S-60-10 or with the amino acid sequence SEQ ID NO: 54, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-10 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 54. In some embodiments, in in total, from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-10 or the amino acid sequence of SEQ ID NO: 54. In some embodiments, the substitutions, insertions, or deletions are located in areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-10 or SEQ ID NO: 54, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-10 antibody, (b) the HVR-H2 amino acid sequence of the S-60-10 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-10. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-10, or with the amino acid sequence of SEQ ID NO: 57, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind to sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6010 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 57. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-10 or the amino acid sequence of SEQ ID NO: 57. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-10 or SEQ ID NO: 57, including post-translational modifications thereof

- 37 046146 sequences. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of the S-60-10 antibody, (b) the HVR-L2 amino acid sequence of the S-60-10 antibody, and (c ) amino acid sequence of HVR-L3 antibody S60-10.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 139. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a light chain comprising the amino acid sequence of SEQ ID NO: 139.

(2) S-60-11.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-11 or the amino acid sequence of SEQ ID NO: 54; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-11 or with the amino acid sequence of SEQ ID NO: 58. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-11 or with the amino acid sequence SEQ ID NO: 54, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-11. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-11 or with amino acid sequence of SEQ ID NO: 58, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-11. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-11 or with the amino acid sequence SEQ ID NO: 54, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-11 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 54. In some embodiments, in a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-11 or the amino acid sequence of SEQ ID NO: 54. In some embodiments, the substitutions, insertions, or deletions are located at areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-11 or SEQ ID NO: 54, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-11 antibody, (b) the HVR-H2 amino acid sequence of the S-60-11 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-11. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical

- 38 046146 with the amino acid sequence of the light chain variable domain of the antibody S-60-11 or with the amino acid sequence of SEQ ID NO: 58, and containing substitutions (for example, conservative substitutions, insertions or deletions compared with the reference sequence), but an anti-sortilin antibody, containing such a sequence retains the ability to bind to sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6011 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 58. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-11 or the amino acid sequence of SEQ ID NO: 58. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-11 or SEQ ID NO: 58, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of antibody S-60-11, (b) the HVR-L2 amino acid sequence of antibody S-60-11, and (c ) amino acid sequence of HVR-L3 antibody S60-11.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a light chain comprising the amino acid sequence of SEQ ID NO: 140.

(3) S-60-12.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-12 or the amino acid sequence of SEQ ID NO: 54; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-12 or with the amino acid sequence of SEQ ID NO: 59. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-12 or with the amino acid sequence SEQ ID NO: 54, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-12. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-12 or with amino acid sequence of SEQ ID NO: 59, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-12. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-12 or with the amino acid sequence SEQ ID NO: 54, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-12 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 54. In some embodiments, in total, from 1 to 5

- 39,046,146 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-12 or the amino acid sequence of SEQ ID NO: 54. In some embodiments, the substitutions, insertions, or deletions are located in regions outside the HVR ( i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-12 or SEQ ID NO: 54, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-12 antibody, (b) the HVR-H2 amino acid sequence of the S-60-12 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-12. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-12, or with the amino acid sequence of SEQ ID NO: 59, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind to sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6012 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 59. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-12 or the amino acid sequence of SEQ ID NO: 59. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-12 or SEQ ID NO: 59, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of the S-60-12 antibody, (b) the HVR-L2 amino acid sequence of the S-60-12 antibody, and (c ) amino acid sequence of HVR-L3 antibody S60-12.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 141. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a light chain comprising the amino acid sequence of SEQ ID NO: 141.

(4) S-60-13.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-13 or the amino acid sequence of SEQ ID NO: 55; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-13 or with the amino acid sequence of SEQ ID NO: 57. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-13 or with the amino acid sequence SEQ ID NO: 55, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-13. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-13 or with aminokis

- 40 046146 lot sequence SEQ ID NO: 57, wherein the light chain variable domain contains the amino acid sequences HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-13. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-13 or with the amino acid sequence SEQ ID NO: 55, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-13 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 55. In some embodiments, in a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-13 or the amino acid sequence of SEQ ID NO: 55. In some embodiments, the substitutions, insertions, or deletions are located at areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-13 or SEQ ID NO: 55, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-13 antibody, (b) the HVR-H2 amino acid sequence of the S-60-13 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-13. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-13, or with the amino acid sequence of SEQ ID NO: 57, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6013 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 57. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-13 or the amino acid sequence of SEQ ID NO: 57. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-13 or SEQ ID NO: 57, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of antibody S-60-13, (b) the HVR-L2 amino acid sequence of antibody S-60-13, and (c ) amino acid sequence of HVR-L3 antibody S60-13.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 135 or SEQ ID NO: 136. In some embodiments, the anti-sortilin antibodies of the present invention contain a light chain comprising the amino acid sequence of SEQ ID NO: 139. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 135 or SEQ ID NO: 136 and a light chain comprising the amino acid sequence of SEQ ID NO: 139.

(5) S-60-14.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-14 or the amino acid sequence of SEQ ID NO: 55; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-14 or with amino

- 41 046146 acid sequence SEQ ID NO: 58. In some embodiments, the anti-sortilin antibodies of this invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable heavy chain domain of antibody S-60-14 or the amino acid sequence of SEQ ID NO: 55, wherein the heavy chain variable domain comprises the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-14. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-14 or with amino acid sequence of SEQ ID NO: 58, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-14. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-14 or with the amino acid sequence SEQ ID NO: 55, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-14 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 55. In some embodiments, in in total, from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-14 or the amino acid sequence of SEQ ID NO: 55. In some embodiments, the substitutions, insertions, or deletions are located in areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-14 or SEQ ID NO: 55, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of antibody S-60-14, (b) the HVR-H2 amino acid sequence of antibody S-60-14, and (c ) amino acid sequence of the HVR-H3 antibody S-60-14. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-14, or with the amino acid sequence of SEQ ID NO: 58, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6014 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 58. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-14 or the amino acid sequence of SEQ ID NO: 58. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-14 or SEQ ID NO: 58, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of antibody S-60-14, (b) the HVR-L2 amino acid sequence of antibody S-60-14, and (c ) amino acid sequence of HVR-L3 antibody S60-14.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 135 or SEQ ID NO: 136. In some embodiments, the anti-sortilin antibodies of the present invention contain a light chain comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 135 or SEQ ID NO: 136 and a light chain comprising

- 42 046146 amino acid sequence SEQ ID NO: 140.

(6) S-60-15.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-15 or the amino acid sequence of SEQ ID NO: 56; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-15 or with the amino acid sequence of SEQ ID NO: 57. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-15 or with the amino acid sequence SEQ ID NO: 56, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-15. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-15 or with amino acid sequence of SEQ ID NO: 57, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-15. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-15 or with the amino acid sequence SEQ ID NO: 56, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-15 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 56. In some embodiments, in in total, from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-15 or the amino acid sequence of SEQ ID NO: 56. In some embodiments, the substitutions, insertions, or deletions are located in areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-15 or SEQ ID NO: 56, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-15 antibody, (b) the HVR-H2 amino acid sequence of the S-60-15 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-15. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-15, or with the amino acid sequence of SEQ ID NO: 57, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6015 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 57. In some embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the antibody S-60-15 light chain variable domain or the amino acid sequence of SEQ ID NO: 57. In some embodiments, the substitution, insertion, or deletion is

- 43 046146 wives in areas outside of HVR (i.e. FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-15 or SEQ ID NO: 57, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of antibody S-60-15, (b) the HVR-L2 amino acid sequence of antibody S-60-15, and (c ) amino acid sequence of HVR-L3 antibody S60-15.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 139. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a light chain comprising the amino acid sequence of SEQ ID NO: 139.

(7) S-60-15.1.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-15.1 or the amino acid sequence of SEQ ID NO: 56; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-15.1 or with the amino acid sequence of SEQ ID NO: 60. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-15.1 or with the amino acid sequence SEQ ID NO: 56, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-15.1. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-6015.1 or with the amino acid sequence SEQ ID NO: 60, wherein the light chain variable domain contains the amino acid sequences HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-15.1. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-15.1 or with the amino acid sequence SEQ ID NO: 56, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S60-15.1 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 56. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-15.1 or the amino acid sequence of SEQ ID NO: 56. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-15.1 or SEQ ID NO: 56, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-15.1 antibody, (b) the HVR-H2 amino acid sequence of the S-60-15.1 antibody, and (c ) amino acid sequence of HVR-H3 antibody S-60-15.1. In some embodiments, anti-sortilin antibodies according to this

- 44 046146 of the invention contain a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95% , at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-15.1 or with the amino acid sequence of SEQ ID NO: 60, and contains substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-15.1 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 60. In some embodiments, in a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-15.1 or the amino acid sequence of SEQ ID NO: 60. In some embodiments, the substitutions, insertions, or deletions are located at areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-15.1 or SEQ ID NO: 60, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVRL1 amino acid sequence of antibody S-60-15.1, (b) the HVR-L2 amino acid sequence of antibody S-60-15.1, and (c) the HVR-L2 amino acid sequence of antibody S-60-15.1. HVR-L3 sequence of antibody S-60-15.1.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 142. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a light chain comprising the amino acid sequence of SEQ ID NO: 142.

(8) S-60-16.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-16 or the amino acid sequence of SEQ ID NO: 56; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-16 or with the amino acid sequence of SEQ ID NO: 77. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-16 or with the amino acid sequence SEQ ID NO: 56, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-16. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-16 or with amino acid sequence of SEQ ID NO: 77, wherein the light chain variable domain contains the amino acid sequences HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-16. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-16 or with the amino acid sequence SEQ ID NO: 56, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some implementations

- 45,046,146 tions, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-16 or the amino acid sequence of SEQ ID NO: 56. In some embodiments, in a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-16 or the amino acid sequence of SEQ ID NO: 56. In some embodiments, the substitutions, insertions, or deletions are located at areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-16 or SEQ ID NO: 56, including post-translational modifications of this sequence. In a specific embodiment, the VH contains one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-16 antibody, (b) the HVR-H2 amino acid sequence of the S-60-16 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-16. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-16, or with the amino acid sequence of SEQ ID NO: 77, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind to sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6016 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 77. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-16 or the amino acid sequence of SEQ ID NO: 77. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-16 or SEQ ID NO: 77, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of antibody S-60-16, (b) the HVR-L2 amino acid sequence of antibody S-60-16, and (c ) amino acid sequence of HVR-L3 antibody S60-16.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 131. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a light chain comprising the amino acid sequence of SEQ ID NO: 131.

(9) S-60-18.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-18 or the amino acid sequence of SEQ ID NO: 56; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-18 or with the amino acid sequence of SEQ ID NO: 78. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-18 or with the amino acid sequence SEQ ID NO: 56, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-18. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least

- 46 046146 at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-18 or with the amino acid sequence of SEQ ID NO: 78, wherein the variable light chain domain contains the amino acid sequences HVR-L1, HVR-L2 and HVR-L3 of antibody S-60 -18. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-18 or with the amino acid sequence SEQ ID NO: 56, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-18 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 56. In some embodiments, in a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-18 or the amino acid sequence of SEQ ID NO: 56. In some embodiments, the substitutions, insertions, or deletions are located in areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-18 or SEQ ID NO: 56, including post-translational modifications of this sequence. In a specific embodiment, the VH contains one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-18 antibody, (b) the HVR-H2 amino acid sequence of the S-60-18 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-18. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-18, or with the amino acid sequence of SEQ ID NO: 78, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind to sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6018 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 78. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-18 or the amino acid sequence of SEQ ID NO: 78. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-18 or SEQ ID NO: 78, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of the S-60-18 antibody, (b) the HVR-L2 amino acid sequence of the S-60-18 antibody, and (c ) amino acid sequence of HVR-L3 antibody S60-18.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a light chain comprising the amino acid sequence of SEQ ID NO: 132.

(10) S-60-19.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-19 or the amino acid sequence of SEQ ID NO: 54; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%,

- 47 046146 at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-19 or with the amino acid sequence of SEQ ID NO: 79. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence of at least at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98 %, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-19 or with the amino acid sequence of SEQ ID NO: 54, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR- H2 and HVR-H3 antibodies S-60-19. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-19 or with amino acid sequence of SEQ ID NO: 79, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-19. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of antibody S-60-19 or with the amino acid sequence SEQ ID NO: 54, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-19 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 54. In some embodiments, in in total, from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-19 or the amino acid sequence of SEQ ID NO: 54. In some embodiments, the substitutions, insertions, or deletions are located in areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-19 or SEQ ID NO: 54, including post-translational modifications of this sequence. In a specific embodiment, the VH contains one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of the S-60-19 antibody, (b) the HVR-H2 amino acid sequence of the S-60-19 antibody, and (c ) amino acid sequence of the HVR-H3 antibody S-60-19. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-19, or with the amino acid sequence of SEQ ID NO: 79, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6019 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 79. In some embodiments, a total of , from 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the light chain variable domain of antibody S-60-19 or the amino acid sequence of SEQ ID NO: 79. In some embodiments, the substitutions, insertions, or deletions are located in regions outside HVR (i.e. in FR sections). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-19 or SEQ ID NO: 79, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of the S-60-19 antibody, (b) the HVR-L2 amino acid sequence of the S-60-19 antibody, and (c ) amino acid sequence of the HVR-L3 antibody S60-19.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106. In some embodiments, the anti-sortilin antibodies of the present invention contain a heavy chain

- 48 046146 heavy chain containing the amino acid sequence of SEQ ID NO: 133. In some embodiments, the anti-sortilin antibodies of this invention comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a light chain containing the amino acid sequence SEQ ID NO: 133.

(11) S-60-24.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the variable the antibody heavy chain domain S60-24 or the amino acid sequence of SEQ ID NO: 56; and/or the light chain variable domain contains an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-24 or with the amino acid sequence of SEQ ID NO: 80. In some In embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable domain comprising an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of the antibody S-60-24 or with the amino acid sequence SEQ ID NO: 56, wherein the heavy chain variable domain contains the amino acid sequences HVR-H1, HVR-H2 and HVR-H3 of antibody S-60-24. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain comprising an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94% , at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-24 or with amino acid sequence of SEQ ID NO: 80, wherein the light chain variable domain contains the amino acid sequences of HVR-L1, HVR-L2 and HVR-L3 of antibody S-60-24. In some embodiments, the anti-sortilin antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the heavy chain variable domain of the antibody S-60-24 or with the amino acid sequence SEQ ID NO: 56, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but an anti-sortilin antibody containing such a sequence retains the ability to bind sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-60-24 antibody heavy chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 56. In some embodiments, in a total of 1 to 5 amino acids have been substituted, inserted, and/or deleted in the amino acid sequence of the heavy chain variable domain of antibody S-60-24 or the amino acid sequence of SEQ ID NO: 56. In some embodiments, the substitutions, insertions, or deletions are located at areas outside of HVR (i.e. in FR areas). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VH sequence of antibody S-60-24 or SEQ ID NO: 56, including post-translational modifications of this sequence. In a specific embodiment, the VH comprises one, two, or three HVRs selected from: (a) the HVR-H1 amino acid sequence of antibody S-60-24, (b) the HVR-H2 amino acid sequence of antibody S-60-24, and (c ) amino acid sequence of the HVR-H3 antibody S-60-24. In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable domain (VL) sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94 %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of the light chain variable domain of antibody S-60-24, or with the amino acid sequence of SEQ ID NO: 80, and containing substitutions (eg, conservative substitutions, insertions or deletions compared to the reference sequence), but the anti-sortilin antibody containing such sequence retains the ability to bind to sortilin. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and/or deleted in the S-6024 antibody light chain variable domain amino acid sequence or amino acid sequence SEQ ID NO: 80. In some embodiments, in approximately

- 49 046146 of greater complexity, from 1 to 5 amino acids have been substituted, inserted and/or deleted in the amino acid sequence of the light chain variable domain of the antibody S-60-24 or the amino acid sequence SEQ ID NO: 80. In some embodiments, the substitution, insertion or deletions are located in regions outside the HVR (i.e., FR regions). In some embodiments, the substitutions, insertions, or deletions are located at FR regions. Optionally, the anti-sortilin antibody contains the VL sequence of antibody S-60-24 or SEQ ID NO: 80, including post-translational modifications of this sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) the HVR-L1 amino acid sequence of antibody S-60-24, (b) the HVR-L2 amino acid sequence of antibody S-60-24, and (c ) amino acid sequence of HVR-L3 antibody S60-24.

In some embodiments, the anti-sortilin antibodies of the invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138. In some embodiments, the anti-sortilin antibodies of the invention comprise a light chain comprising the amino acid sequence of SEQ ID NO: 134. In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a light chain comprising the amino acid sequence of SEQ ID NO: 134.

In some embodiments, the anti-sortilin antibody of the present invention binds substantially the same sortilin epitope as an antibody comprising a heavy chain variable domain and a light chain variable domain of an antibody selected from the group consisting of S-60-10, S-60 -11, S-6012, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-16, S-60- 18, S-60-19, and S-60-24.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-10. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same epitope of sortilin as S-60-10. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-10. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-10. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6010.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-11. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-11. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-11. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-11. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6011.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-12. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same epitope of sortilin as S-60-12. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-12. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-12. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6012.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-13. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-13. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-13. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-13. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6013.

In some embodiments, the anti-sortilin antibody is a monoclonal

- 50 046146 new antibody against sortilin S-60-14. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-14. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-14. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-14. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6014.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-15. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-15. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-15. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-15. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6015.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-15.1. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-15.1. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-6015.1. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-15.1. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-60-15.1.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-16. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same epitope of sortilin as S-60-16. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-16. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-16. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6016.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-18. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-18. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-18. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-18. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6018.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-19. In some embodiments, the anti-sortilin antibody is an isolated antibody that binds substantially the same sortilin epitope as S-60-19. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-19. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-19. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6019.

In some embodiments, the anti-sortilin antibody is an anti-sortilin monoclonal antibody S-60-24. In some embodiments, an anti-sortilin antibody is an isolated antibody that binds substantially the same epitope of sortilin as

- 51 046146

S-60-24. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the heavy chain variable region of the monoclonal antibody S-60-24. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising the light chain variable region of the monoclonal antibody S-60-24. In some embodiments, the anti-sortilin antibody is an isolated antibody comprising a heavy chain variable region and a light chain variable region of the monoclonal antibody S-6024.

In some embodiments, the anti-sortilin antibody is an antagonist antibody. In some embodiments, the anti-sortilin antibody is an agonistic antibody. In some embodiments, the anti-sortilin antibodies of the present invention are of the IgG class, the IgM class, or the IgA class. In some embodiments, the anti-sortilin antibodies of the present invention are of the IgG class and are of the IgG1, IgG2, IgG3, or IgG4 isotype.

Additional anti-sortilin antibodies, for example, antibodies that specifically bind to the sortilin protein of the present invention, can be identified, screened, and/or characterized for their physical/chemical properties and/or biological activities by various assays known in the art.

Certain aspects of the present invention relate to the use of two or more anti-sortilin antibodies that, when used together, exhibit additive or synergistic effects compared to the use of the corresponding individual anti-sortilin antibody.

In some embodiments, the anti-sortilin antibody of the present invention is an antibody fragment that binds to the human sortilin protein.

In some embodiments, the anti-sortilin antibody of the present invention is an antibody fragment that binds to one or more human proteins selected from the group consisting of human sortilin, a natural variant of human sortilin, and a disease variant of human sortilin.

In some embodiments, the anti-sortilin antibody of the present invention is an antibody fragment, wherein the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, or scFv fragment.

Antibody scaffolds.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising one or more (e.g., one or more, two or more, three or more, or all four) framework regions selected from VH FR1, VH FR2 , VH FR3, and VH FR4 (as indicated in Tables 20-23). In some embodiments, VH FR1 contains the sequence QVQLQESGPGLVKPSETLSLTCAVSG (SEQ ID NO: 35). In some embodiments, VH FR2 contains the sequence WIRQPPGKGLEWIG (SEQ ID NO: 36). In some embodiments, VH FR3 contains a sequence corresponding to Formula VI: XjVTISVDTSKNQFSLX ₂ LSSVTAADTAVYYC (SEQ ID NO: 39), where X1 is Q or R, and X ₂ is E or K. In some embodiments, VH FR3 contains a sequence selected from the group consisting of SEQ ID NO: 37-38. In some embodiments, VH FR4 contains the sequence WGQGTTVTVSS (SEQ ID NO: 40). In some embodiments, the antibody comprises a heavy chain variable region comprising VHFR1 containing the sequence of SEQ ID NO: 35, VH FR2 containing the sequence of SEQ ID NO: 36, VH FR3 in accordance with Formula VI, and VH FR4 containing the sequence SEQ ID NO: 40.

In some embodiments, the antibody comprises a heavy chain variable region comprising VH FR1 comprising the sequence of SEQ ID NO: 35, VH FR2 containing the sequence of SEQ ID NO: 36, VH FR3 containing the sequence selected from SEQ ID NO: 37-38, and VH FR4 containing the sequence SEQ ID NO: 40.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising VH FR1, VH FR2, VH FR3, and VH FR4 antibodies S-6010, S-60-11, S-60-12, S-60 -13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S- 60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-6015.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as indicated in Table 20-23).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising one or more (e.g., one or more, two or more, three or more, or all four) framework regions selected from VL FR1, VL FR2 , VL FR3, and VL FR4 (as indicated in Tables 24-27). In some embodiments, VL FR1 contains the sequence according to Formula VII: DIVMTQSPLSLPVTPGXiX ₂ ASISC (SEQ ID NO: 44), where X1 is E or G, and X ₂ is P or S. In some embodiments, VL FR1 contains a sequence selected from the group consisting of SEQ ID NO: 41-43. In some

- 52 046146 embodiments, VL FR2 contains a sequence in accordance with formula VIII: WYLQKPGQX ₁ PQLLIY (SEQ ID NO: 47), where X ₁ represents S or P. In some implementations, VL FR2 contains a sequence selected from the group, consisting of SEQ ID NO: 4546. In some embodiments, VL FR3 contains the sequence in accordance with Formula IX: GVPDRX1SGSGSGT DFTLKISRX2EAEDVGX3YYC (SEQ ID NO: 52), where X1 is F or L, X ₂ is A or V, and X ₃ represents V or A. In some embodiments, VL FR3 contains a sequence selected from the group consisting of SEQ ID NO: 48-51. In some embodiments, VL FR4 contains the sequence FGGGTKVEIK (SEQ ID NO: 53). In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising VL FR1 containing a Formula VII sequence, VL FR2 containing a Formula VIII sequence, VL FR3 containing a Formula IX sequence, and VL FR4 containing the sequence SEQ ID NO: 53.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising VL FR1 comprising a sequence selected from SEQ ID NOs: 41-43, VL FR2 comprising a sequence selected from SEQ ID NOs: 45-46, VL FR3 containing the sequence selected from SEQ ID NO: 48-51, and VL FR4 containing the sequence SEQ ID NO: 53.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a light chain variable region comprising VL FR1, VL FR2, VL FR3, and VL FR4 antibodies S-60-10, S-60-11, S-60-12, S -60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-6015.9 [N33Y ], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60 -15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as indicated in Tables 24-27).

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising one or more (e.g., one or more, two or more, three or more, or all four) framework regions selected from VH FR1, VH FR2 , VH FR3, and VH FR4 (as defined in Tables 20-23), and a light chain variable region comprising one or more (e.g., one or more, two or more, three or more, or all four) framework regions, selected from VL FR1, VL FR2, VL FR3, and VL FR4 (as indicated in Tables 24-27). In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising a VH FR1 containing the sequence of SEQ ID NO: 35, a VH FR2 containing the sequence of SEQ ID NO: 36, a VH FR3 in accordance with Formula VI, and a VH FR4 containing the sequence SEQ ID NO: 40; and a light chain variable region comprising VL FR1 containing the sequence corresponding to Formula VII, VL FR2 containing the sequence corresponding to Formula VIII, VL FR3 containing the sequence corresponding to Formula IX, and VL FR4 containing the sequence SEQ ID NO: 53. B In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising VH FR1 comprising the sequence of SEQ ID NO: 35, VH FR2 containing the sequence of SEQ ID NO: 36, VH FR3 containing the sequence selected from SEQ ID NO: 37-38, and VH FR4 containing the sequence SEQ ID NO: 40; a light chain variable region comprising VL FR1 containing a sequence selected from SEQ ID NO: 41-43, VL FR2 containing a sequence selected from SEQ ID NO: 45-46, VL FR3 containing a sequence selected from SEQ ID NO: 48-51, and VL FR4 containing the sequence SEQ ID NO: 53.

In some embodiments, the anti-sortilin antibodies of the present invention comprise a heavy chain variable region comprising VH FR1, VH FR2, VH FR3, and VH FR4 antibodies S-6010, S-60-11, S-60-12, S-60 -13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S- 60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-6015.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as specified in Table 20-23), and a light chain variable region containing VL FR1, VL FR2, VL FR3, and VL FR4 antibodies S -60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-6015.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K ], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60 -15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S60-19, or S-60-24 (as indicated in Table 24-27).

Antibody activity against sortilin.

In certain aspects of any of the anti-sortilin antibodies, the anti-sortilin antibodies of this invention may inhibit one or more activities of the sortilin protein, including, without limitation, reducing cellular levels of sortilin (e.g., cell surface sortilin levels, intracellular sortilin levels, and/or total levels sortilina); increasing pro levels

- 53 046146 granulin (for example, extracellular levels of progranulin and/or cellular levels of progranulin); and inhibition of interaction (eg, binding) between progranulin and sortilin. As provided herein, the anti-sortilin antibodies of the present invention may inhibit additional activities of the sortilin protein, including, without limitation, inhibition of interaction (e.g., binding) with one or more proneurotrophins of the present invention (proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, etc.), neurotrophins according to this invention (neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, sortilin propeptide (Sort-pro) , amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), and receptor associated protein (RAP), decreased PCSK9 secretion, deacreasing production of beta-amyloid peptide.

In some embodiments, the invention provides an anti-sortilin antibody, wherein (a) the anti-sortilin antibody increases extracellular levels of progranulin, decreases cellular levels of sortilin, inhibits the interaction between sortilin and progranulin, or any combination thereof; (b) an anti-sortilin antibody reduces cell surface levels of sortilin, increases extracellular levels of progranulin, inhibits the interaction between sortilin and progranulin, or any combination thereof; (c) an anti-sortilin antibody reduces cell surface sortilin levels, reduces intracellular sortilin levels, reduces total sortilin levels, or any combination thereof; (d) the anti-sortilin antibody induces sortilin degradation, sortilin cleavage, sortilin internalization, decreased sortilin expression, or any combination thereof; (e) anti-sortilin antibody reduces cellular levels of sortilin and inhibits the interaction between sortilin and progranulin; (f) anti-sortilin antibody reduces cellular levels of sortilin and increases cellular levels of progranulin; and/or (g) the anti-sortilin antibody increases the effective concentration of progranulin.

In some embodiments, the invention provides an anti-sortilin antibody, wherein the anti-sortilin antibody reduces cell surface levels of sortilin, increases extracellular levels of progranulin, inhibits the interaction between sortilin and progranulin, or any combination thereof.

In some embodiments, the anti-sortilin antibody of the present invention (a) reduces cell surface levels of sortilin with a half-maximal effective concentration (EC ₅₀ ) of less than 150 pM, as measured by flow cytometry; (b) reduces cell surface sortilin levels by greater than about 50% at 1.25 nM IgG, by greater than about 80% at 0.63 nM IgG, or by greater than about 69% at 150 nM IgG compared to control, when measured by flow cytometry; increases the secretion of progranulin by more than about 1.13 times compared to the control at 0.63 nM IgG, or more than about 1.22 times compared to the control at 50 nM IgG, as measured by standard ELISA; blocks the binding of progranulin to sortilin with a half-maximal effective concentration (EC50) of less than 0.325 nM, as measured by flow cytometry; (e) blocks the binding of progranulin to sortilin by more than about 88% at 50 nM IgG, or by more than about 27.5% at 150 nM IgG, compared to control, as measured by flow cytometry; or (f) any combination thereof (see, for example, examples 2-4).

In some embodiments, the anti-sortilin antibody of the invention (a) reduces cell surface levels of sortilin with a half-maximal effective concentration (EC50) of less than 681 pM as measured by flow cytometry; (b) reduces cell surface sortilin levels by greater than about 40% at 1.25 nM IgG, by greater than about 29% at 0.6 nM IgG, or by greater than about 62% at 150 nM IgG compared to control, when measured by flow cytometry; (c) increases the secretion of progranulin by more than about 1.11 times that of the 0.63 nM IgG control, or by more than about 1.75 times that of the 50 nM IgG control, as measured by a standard ELISA; (d) blocks the binding of progranulin to sortilin with a half-maximal effective concentration (EC ₅₀ ) of less than 0.751 nM, as measured by flow cytometry; (e) blocks the binding of progranulin to sortilin by more than about 90% at 50 nM IgG, or by more than about 95% at 150 nM IgG, compared to control, as measured by flow cytometry; or (f) provides any combination thereof (see, for example, Examples 2-4).

Decreased sortilin levels.

In some embodiments, anti-sortilin antibodies of the invention bind to the cell surface expressed sortilin protein of the invention and modulate (e.g., induce or inhibit) one or more activities of the sortilin of the invention upon binding to the surface expressed sortilin protein.

In some embodiments, the anti-sortilin antibodies of the present invention reduce cellular levels of sortilin in vitro. In some embodiments, the anti-sortilin antibodies of the present invention can reduce cellular levels of sortilin in vivo (e.g., in the brain,

- 54 046146 and/or peripheral organs of the individual). In some embodiments, reducing cellular levels of sortilin includes reducing levels of cell surface sortilin. As used herein, an anti-sortilin antibody reduces cell surface sortilin levels if it induces a decrease at the saturating concentration of the antibody (eg, 0.6 nM, 0.63 nM, 1.25 nM, 50 nM, or 150 nM) and/ or compared to a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60) cell surface sortilin levels as measured by any of the in vitro cellular assays or suitable in vivo model described in this document or known to those skilled in the art. In some embodiments, reducing cellular levels of sortilin includes reducing intracellular levels of sortilin. As provided herein, an anti-sortilin antibody reduces intracellular sortilin levels if it induces a decrease at the saturating concentration of the antibody and/or relative to a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S -60) intracellular levels of sortilin when measured using any in vitro cell assay or suitable in vivo model described herein or known to those skilled in the art. In some embodiments, reducing cellular levels of sortilin includes reducing overall levels of sortilin. As provided herein, an anti-sortilin antibody reduces overall sortilin levels if it induces a decrease at the saturating concentration of the antibody and/or relative to a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S -60) total sortilin levels when measured using any in vitro cell assay or suitable in vivo model described herein or known to those skilled in the art.

As used herein, levels of sortilin may refer to levels of expression of the gene encoding sortilin; to the expression levels of one or more transcripts encoding sortilin; to sortilin protein expression levels; and/or to the amount of sortilin protein present inside cells and/or on the surface of cells. Any method known to those skilled in the art for measuring levels of gene expression, transcription, translation, and/or relative protein abundance or localization can be used to determine levels of sortilin.

Cellular sortilin levels can refer to, without limitation, cell surface sortilin levels, intracellular sortilin levels, and total sortilin levels. In some embodiments, reducing cellular levels of sortilin includes reducing levels of cell surface sortilin. In some embodiments, anti-sortilin antibodies of the present invention that reduce cellular levels of sortilin (eg, cell surface levels of sortilin) have one or more of the following characteristics: (1) inhibit or reduce one or more activities of sortilin; (2) the ability to inhibit or reduce the binding of sortilin to one or more of its ligands; (3) the ability to downregulate sortilin expression in sortilin-expressing cells; (4) the ability to interact with, bind, or recognize the protein sortilin; (5) the ability to specifically interact with or bind to the protein sortilin; and (6) the ability to cure, ameliorate, or prevent any aspect of the disease or disorder described or contemplated herein.

In some embodiments, an isolated anti-sortilin antibody of the present invention induces a decrease in sortilin expression. In some embodiments, the isolated anti-sortilin antibody of the present invention induces the cleavage of sortilin. In some embodiments, the isolated anti-sortilin antibody of the present invention induces the internalization of sortilin. In some embodiments, an isolated anti-sortilin antibody of the present invention induces sortilin shedding. In some embodiments, an isolated anti-sortilin antibody of the present invention induces degradation of sortilin. In some embodiments, an isolated anti-sortilin antibody of the present invention induces sortilin desensitization. In some embodiments, an isolated anti-sortilin antibody of the present invention acts as a ligand mimetic to transiently activate sortilin. In some embodiments, an isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing a decrease in cellular levels of sortilin and/or inhibiting the interaction (eg, binding) between sortilin and one or more sortilin ligands. In some embodiments, an isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing degradation of sortilin. In some embodiments, an isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing sortilin cleavage. In some embodiments, the isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing internalization of sortilin. In some embodiments, the isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing sortilin shedding. In some

- 55 046146 embodiments, the isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing a decrease in sortilin expression. In some embodiments, the isolated anti-sortilin antibody of the present invention acts as a ligand mimetic and transiently activates sortilin before inducing sortilin desensitization.

In some embodiments, the anti-sortilin antibodies of the present invention can reduce cellular sortilin levels (eg, cell surface sortilin levels, intracellular sortilin levels, and/or total sortilin levels) by inducing sortilin degradation. Accordingly, in some embodiments, the anti-sortilin antibodies of the present invention induce degradation of sortilin.

The anti-sortilin antibodies of the present invention can reduce cellular levels (eg, cell surface levels) of sortilin with a half-maximal effective concentration (EC50) (eg, when measured in vitro) in the picomolar range of values. In some embodiments, the EC50 of the antibody is less than about 680.9 pM. In some embodiments, the EC50 of the antibody is from about 72.58 pM to about 680.9 nM. In some embodiments, the EC50 of the antibody is from about 103.6 pM to about 680.9 nM. In some embodiments, the EC ₅₀ of the antibody is less than about 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, 100 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 1 pM, or 0.5 PM.

In some embodiments, the EC ₅₀ of the antibody is less than about or equal to about 675 pM, 650 pM, 625 pM, 600 pM, 575 pM, 550 pM, 525 pM, 500 pM, 475 pM, 450 pM, 425 pM , 400 pM, 375 pM, 350 pM, 325 pM, 300 pM, 275 pM, 250 pM, 225 pM, 200 pM, 175 pM, 150 pM, 125 pM, 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1 pM or 0.5 pM.

In some embodiments, the EC50 of the antibody is less than about 680.9 pM. In some embodiments, the EC50 of the antibody is greater than about or equal to about 0.1 pM, 0.5 pM, 1 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM , 80 pM, 90 pM, 100 pM, 125 pM, 150 pM, 175 pM, 200 pM, 225 pM, 250 pM, 275 pM, 300 pM, 325 pM, 350 pM, 375 pM, 400 pM, 425 pM, 450 pM, 475 pM, 500 pM, 525 pM, 550 pM, 575 pM, 600 pM, 625 pM, 650 pM, 675 pM. In other words, the EC ₅₀ of an antibody may be any value within a range having an upper limit of about 675 pM, 650 nM, 650 pM, 625 pM, 600 pM, 575 pM, 550 pM, 525 pM, 500 pM, 475 pM, 450 pM, 425 pM, 400 pM, 375 pM, 350 pM, 325 pM, 300 pM, 275 pM, 250 pM, 225 pM, 200 pM, 175 pM, 150 pM, 125 pM, 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 1 pM, or 0.5 pM, and an independently selected lower limit of approximately 0.1 pM, 0.5 pM, 1 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 125 pM, 150 pM, 175 pM, 200 pM, 225 pM, 250 pM, 275 pM at In this case, the lower limit has less significance than the upper limit. In some embodiments, the EC50 of the antibody is any of about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 15 pM, 20 pM, 25 pM , 30 pM, 35 pM, 40 pM, 45 pM, 50 pM, 55 pM, 60 pM, 65 pM, 70 pM, 75 pM, 80 pM, 85 pM, 90 pM, 95 pM, 100 pM, 105 pM, 110 pm, 115 pm, 120 pm, 125 pm, 130 pm, 135 pm, 140 pm, 145 pm, 150 pm, 155 pm, 160 pm, 165 pm, 170 pm, 175 pm, 180 pm, 185 pm, 190 pm, 195 pM or 200 pM.

In some embodiments, the anti-sortilin antibody of the present invention reduces cell surface levels of sortilin with a half-maximal effective concentration (EC50) of less than 150 pM as measured by flow cytometry. In some embodiments, the EC50 of the anti-sortilin antibody of this invention is about 103.6 pM. In some embodiments, the EC50 of the anti-sortilin antibody of the invention is about 72.58 pM.

In some embodiments, the anti-sortilin antibody of the present invention reduces cell surface sortilin levels by more than about 40% at 1.25 nM IgG, or by more than about 80% at 0.63 nM IgG, as measured by flow cytometry. In some embodiments, the anti-sortilin antibody of the present invention reduces cell surface sortilin levels by about 60.92% at 1.25 nM IgG, as measured by flow cytometry. In some embodiments, the anti-sortilin antibody of the present invention reduces cell surface sortilin levels by about 69.3% at 150 nM IgG, as measured by flow cytometry. In some embodiments, the anti-sortilin antibody of the present invention reduces cell surface sortilin levels by about 70.3% at 150 nM IgG, as measured by flow cytometry.

Various methods are known to those skilled in the art for measuring antibody EC50 values, including, for example, flow cytometry (see, for example, Example 2). In some embodiments, EC ₅₀ is measured in vitro using cells modified to express human sortilin. In some embodiments, EC ₅₀ is measured at a temperature of approximately 4°C. In some embodiments, EC ₅₀ is measured at a temperature of approximately 25°C. In some variants

- 56 046146 implementations, EC ₅₀ is measured at a temperature of approximately 35°C. In some embodiments, EC ₅₀ is measured at a temperature of approximately 37°C. In some embodiments, EC ₅₀ is determined using a monovalent antibody (eg, Fab) or a full-length antibody in monovalent form. In some embodiments, EC ₅₀ is determined using an antibody containing constant regions that exhibit increased Fc receptor binding. In some embodiments, EC ₅₀ is determined using an antibody containing constant regions that exhibit reduced Fc receptor binding.

In some embodiments, the anti-sortilin antibodies of the present invention have superior activity in reducing cell surface sortilin levels compared to a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention reduce cellular levels (e.g., cell surface levels) of sortilin with a lower EC ₅₀ value (e.g., by in vitro measurement) than a control antibody (e.g., an anti-sortilin antibody having a variable region heavy chain and light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention reduce cellular levels (e.g., cell surface levels) of sortilin with an EC ₅₀ that is at least about 5%, at least at least at least at least about approximately approximately approximately

10%, 25%, 40%, 55%, 70%, according to at at at least at least at least at least at least at least at least at least at least at least approximately approximately approximately approximately approximately

15%, 30%, 45%, 60%, 75%, according to at at at least at least at least at least at least at least at least at least at least at least at least approximately approximately approximately approximately approximately

20%, 35%, 50%, 65%, 80%, each at least about 85%, at least about 90%, at least about 95%, or at least about 99% lower than EC ₅₀ of a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention reduce cellular levels (e.g., cell surface levels) of sortilin with an EC ₅₀ that is at least about 1-fold, at least about 1.1-fold, at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 12.5 times, at least about 15 times, at least about 17.5 times, at least about 20 times, at least about 22.5 times, at least about 25 times, at least about 27.5 times, at least about 30 times, at least about 50 times, or at least about 100 times lower than EC ₅₀ control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60).

In some embodiments, the anti-sortilin antibodies of the present invention have an EC ₅₀ that is at least 1.5-fold lower than that of a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S -60). In some embodiments, the anti-sortilin antibodies of the present invention have an EC ₅₀ that is at least 1.1-fold lower than that of a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S -60).

In some embodiments, the anti-sortilin antibody of the present invention (a) reduces cell surface levels of sortilin with a half-maximal effective concentration (EC ₅₀ ) of less than 681 pM, as measured by flow cytometry; (b) reduces cell surface sortilin levels by more than about 40% at 1.25 nM IgG, by more than about 29% at 0.6 nM IgG, or by more than about 62% at 150 nM IgG, compared to control , when measured by flow cytometry; (c) increases the secretion of progranulin by more than about 1.11 times that of the 0.63 nM IgG control, or by more than about 1.75 times that of the 50 nM IgG control, as measured by a standard ELISA; (d) blocks the binding of progranulin to sortilin with a half-maximal effective concentration (EC ₅₀ ) of less than 0.751 nM, as measured by flow cytometry; (e) blocks the binding of progranulin to sortilin by more than about 90% at 50 nM IgG, or by more than about 95% at 150 nM IgG, compared to control, as measured by flow cytometry; or (f) provides any combination thereof (see, for example, Examples 2-4).

Increased levels of progranulin.

In some embodiments, the anti-sortilin antibodies of this invention increase extracellular levels of progranulin in vitro. In some embodiments, the anti-sortilin antibodies of this invention can increase cellular levels of progranulin or in vivo (eg, in the brain, blood, and/or peripheral organs of an individual). As used herein, an anti-sortilin antibody increases extracellular levels of progranulin if it induces an increase at saturating concentrations of the antibody (eg, 0.6 nM, 0.63 nM, 1.25 nM, 50 nM, or 150 nM ) and/or compared to a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60) extracellular levels of progranulin when measured by any in vitro cell-based or tissue-based assays (such as brain) tests described herein or known to those skilled in the art. As provided herein, an anti-sortilin antibody increases cellular progranulin levels if it induces an increase at a saturating antibody concentration (eg, 0.6 nM, 0.63 nM, 1.25 nM, 50 nM, or 150 nM) and/or compared to a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60) cellular progranulin levels when measured by any in vitro cell-based assay or tissue (such as brain tissue) assay described herein or known to those skilled in the art.

As used herein, progranulin levels may refer to expression levels of the gene encoding progranulin; to the expression levels of one or more transcripts encoding progranulin; to progranulin protein expression levels; and/or to the amount of progranulin protein secreted by the cells and/or present in the cells. Any method known to those skilled in the art for measuring levels of gene expression, transcription, translation, relative protein abundance, protein secretion, and/or protein localization can be used to determine progranulin levels.

As used herein, progranulin levels can refer to, without limitation, extracellular progranulin levels, intracellular progranulin levels, and total progranulin levels. In some embodiments, increasing progranulin levels includes increasing extracellular progranulin levels.

In some embodiments, the anti-sortilin antibody of the present invention increases progranulin secretion by more than about 1.11-fold compared to the control at 0.63 nM IgG, as measured by a standard ELISA. In some embodiments, the anti-sortilin antibody of the present invention increases progranulin secretion by approximately 1.42-fold over control at 0.63 nM IgG, as measured by a standard ELISA. In some embodiments, the anti-sortilin antibody of the present invention increases progranulin secretion by more than about 1.75-fold compared to a 50 nM IgG control, as measured by a standard ELISA. In some embodiments, the anti-sortilin antibody of the present invention increases progranulin secretion by approximately 1.97-fold over control at 50 nM IgG, as measured by a standard ELISA. In some embodiments, the anti-sortilin antibody of the present invention increases progranulin secretion by approximately 2.29-fold over control at 50 nM IgG, as measured by a standard ELISA.

Various methods for measuring the secretion of progranulin are known to those skilled in the art, including, for example, by ELISA (see, for example, Examples 2 and 4). In some embodiments, EC ₅₀ is measured in vitro using cells expressing human sortilin. In some embodiments, progranulin secretion is determined using a monovalent antibody (eg, Fab) or a full-length antibody in monovalent form. In some embodiments, progranulin secretion is determined using an antibody containing constant regions that exhibit increased Fc receptor binding. In some embodiments, progranulin secretion is determined using an antibody containing constant regions that exhibit reduced Fc receptor binding.

In some embodiments, the anti-sortilin antibodies of the present invention have superior activity for increasing progranulin levels compared to a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention increase levels (e.g., extracellular levels) of progranulin with a lower EC ₅₀ value (e.g., when measured in vitro) than a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention increase levels (e.g., extracellular levels) of progranulin by at least about 5%, at least about 10%, at least about 15%, at least about about about about

20%, 35%, 50%, 65%, 80%, at at at at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least at least approximately approximately approximately approximately

25%, 40%, 55%, 70%, 85%, according to at at at least at least at least at least at least at least at least at least at least at least at least approximately approximately approximately approximately approximately

30%, 45%, 60%, 75%, 90%, at at least at least at least at least at least or at least about 99% compared to (than) a control antibody (e.g. against sortilin, having a heavy chain variable region and a variable

- 58 046146 light chain region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention increase the levels (e.g., extracellular levels) of progranulin by about 1-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times , at least about 9 times, at least about 10 times, at least about 12.5 times, at least about 15 times, at least about 17.5 times, at least about 20 times, at least about 22.5 times, at least about 25 times, at least about 27.5 times, at least about 30 times, at least about 50 times, or at least about 100 times stronger than a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60).

In some embodiments, the anti-sortilin antibodies of the present invention increase progranulin levels approximately 1.1-fold greater than a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention increase progranulin levels approximately 1.3-fold greater than a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60).

In some embodiments, the anti-sortilin antibodies of the present invention increase the effective concentration of progranulin. The effective concentration of progranulin refers to the concentration of progranulin in plasma or cerebrospinal fluid. In some embodiments, the increase in the effective concentration of progranulin is an increase of greater than 1.5 times. In some embodiments, the effective concentration of progranulin remains elevated for 7-28 days.

Reduced interaction between sortilin and binding partner.

In some embodiments, anti-sortilin antibodies inhibit the interaction (e.g., binding) between the sortilin protein of the invention and one or more proteins selected from progranulin, proneurotrophin, neurotrophin, proneurotrophin-3, neurotrophin-3, proneurotrophin-4/5, neurotrophin -4/5, pro-nerve growth factor (pro-NGF), nerve growth factor (NGF), pro-brain-derived neurotrophic factor (pro-BDNF), brain-derived neurotrophic factor (BDNF), neurotensin, p75, propeptide sortilin (Sort- pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and receptor associated protein (RAP), and/ or natural options. In a specific embodiment, the anti-sortilin antibodies of the present invention inhibit the interaction (eg, binding) between the sortilin protein of the present invention and progranulin.

In some embodiments, anti-sortilin antibodies can inhibit the interaction (e.g., binding) between the sortilin protein of the invention and a neurotrophin of the invention, such as proneurotrophin, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, and BDNF. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and neurotensin. In other embodiments, anti-sortilin antibodies inhibit the interaction (eg, binding) between the sortilin protein of the invention and p75. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and Sort-pro. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and APP. In other embodiments, anti-sortilin antibodies may inhibit the production of beta-amyloid peptide. In other embodiments, anti-sortilin antibodies may inhibit the transport and secretion of PCSK9. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and LpL. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and APOA5. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and APOE. In other embodiments, anti-sortilin antibodies may inhibit the interaction (eg, binding) between the sortilin protein of the invention and RAP.

In some embodiments, the anti-sortilin antibodies of the invention bind to the sortilin protein of the invention expressed on the surface of a cell, and the naked antibodies inhibit the interaction (eg, binding) between the sortilin protein and one or more sortilin ligands. In some embodiments, anti-sortilin antibodies

- 59 046146 of this invention, which bind to the sortilin protein of this invention (of the present), inhibit the interaction (eg, binding) between the sortilin protein and one or more sortilin ligands by reducing the effective levels of sortilin available to interact with these proteins on the surface cells or inside a cell. In some embodiments, anti-sortilin antibodies of the invention that bind to the sortilin protein of the invention inhibit the interaction (eg, binding) between the sortilin protein and one or more sortilin ligands by inducing degradation of sortilin.

A. Reduced interaction between sortilin and progranulin.

In some embodiments, the anti-sortilin antibodies of the present invention increase progranulin levels and/or decrease cellular levels of sortilin while blocking (eg, inhibiting) the interaction (eg, binding) between sortilin and progranulin. Accordingly, in some embodiments, the anti-sortilin antibodies of the present invention block the interaction (eg, binding) between sortilin and progranulin. As used herein, an anti-sortilin antibody blocks the interaction (e.g., binding) between sortilin and progranulin if it reduces the binding of progranulin to sortilin compared to a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region chains corresponding to S-60) at a saturating antibody concentration (e.g., 0.6 nM, 0.63 nM, 1.25 nM, 50 nM, or 150 nM) in any in vitro or cell culture assay described herein or known to specialists.

The anti-sortilin antibodies of the present invention can reduce the binding of progranulin to sortilin with a half-maximal effective concentration (EC50) (eg, when measured in vitro) in the picomolar range of values. In some embodiments, the EC50 of the antibody is less than about 2.2 nM. In some embodiments, the EC50 of the antibody is less than about 1.22 nM. In some embodiments, the EC50 of the antibody is less than about 751 pM. In some embodiments, the EC50 of the antibody is from about 325 pM to about 751 nM. In some embodiments, the EC50 of the antibody is from about 405 pM to about 751 nM. In some embodiments, the EC ₅₀ of the antibody is from about 588 pM to about 751 nM. In some embodiments, the EC ₅₀ of the antibody is less than about 2.2 nM, 2.1 nM, 2.0 nM, 1.9 nM, 1.8 nM, 1.7 nM, 1.6 nM, 1.5 nM , 1.4 nM, 1.3 nM, 1.2 nM, 1.1 nM, 1.0 nM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, 100 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 1 pM or 0.5 pM.

In some embodiments, the EC ₅₀ value of the antibody upon decreasing binding of progranulin to sortilin is less than or equal to about 2.2 nM, 2.1 nM, 2.0 nM, 1.9 nM, 1.8 nM, 1.7 nM , 1.6 nM, 1.5 nM, 1.4 nM, 1.3 nM, 1.2 nM, 1.1 nM, 1.0 nM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM , 475 pM, 450 pM, 425 pM, 400 pM, 375 pM, 350 pM, 325 pM, 300 pM, 275 pM, 250 pM, 225 pM, 200 pM, 175 pM, 150 pM, 125 pM, 100 pM, 90 pm, 80 pm, 70 pm, 60 pm, 50 pm, 40 pm, 30 pm, 20 pm, 10 pm, 9 pm, 8 pm, 7 pm, 6 pm, 5 pm, 4 pm, 3 pm, 2 pm, 1 pM or 0.5 pM.

In some embodiments, the EC50 of the anti-sortilin antibody of this invention is approximately 1.22 nM. In some embodiments, the EC50 of the anti-sortilin antibody of this invention is approximately 588 pM. In some embodiments, the EC50 of the anti-sortilin antibody of this invention is approximately 405 pM. In some embodiments, the EC50 of the anti-sortilin antibody of this invention is approximately 325 pM.

Various methods for measuring antibody EC50 values are known to those skilled in the art, including, for example, flow cytometry (see, for example, Example 3). In some embodiments, the EC ₅₀ of decreased binding of progranulin to sortilin (Sortlin) is measured in vitro using cells expressing human sortilin. In some embodiments, EC50 is measured at a temperature of approximately 4°C. In some embodiments, EC ₅₀ is measured at a temperature of approximately 25°C. In some embodiments, EC ₅₀ is measured at a temperature of approximately 35°C. In some embodiments, EC ₅₀ is measured at a temperature of approximately 37°C. In some embodiments, the EC50 of decreased binding of progranulin to sortilin is determined using a monovalent antibody (eg, Fab) or a full-length antibody in monovalent form. In some embodiments, EC ₅₀ is determined using an antibody containing constant regions that exhibit increased Fc receptor binding. In some embodiments, the EC50 of decreased binding of progranulin to sortilin is determined using an antibody containing constant regions that exhibit decreased Fc receptor binding.

In some embodiments, the anti-sortilin antibodies of the present invention have greater activity in reducing the binding of progranulin to sortilin (Sortlin) compared to a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S- 60). In some embodiments, the anti-sortilin antibodies of the present invention reduce the binding of progranulin to sortilin with a lower EC50 value (e.g., when measured in vitro) than a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a variable region

- 60 046146 light chain blade, corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention reduce the binding of progranulin to sortilin with an EC50 value of at least about 5%, at least about 10%, at least about 15%, at least about 20% , at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, according to at least about 95%, or at least about 99% lower than the EC ₅₀ of a control antibody (eg, an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the present invention reduce the binding of progranulin to sortilin with an EC50 of at least about 1-fold, at least about 1.1-fold, at least about 1.5-fold, at least at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 12.5 times, at least about 15 times, at least about 17.5 times, at least about 20 times, at least about 22.5 times, at least about 25 times, at least about 27.5 times, at least about 30 times, at least about 50 times, or at least about 100 times lower than the EC ₅₀ of a control antibody (e.g., antibody against sortilin having a heavy chain variable region and a light chain variable region corresponding to S-60).

In some embodiments, the anti-sortilin antibodies of the present invention have an EC50 that is at least 1.3-fold lower than that of a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60 ). In some embodiments, the anti-sortilin antibodies of the present invention have an EC ₅₀ that is at least 1.8-fold lower than that of a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S- 60). In some embodiments, the anti-sortilin antibodies of the present invention have an EC50 that is at least 1.9-fold lower than that of a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60 ). In some embodiments, the anti-sortilin antibodies of the present invention have an EC50 that is at least 2.3 times lower than that of a control antibody (e.g., an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60 ).

Any in vitro cell assay or suitable in vivo model described herein or known to those skilled in the art can be used to measure inhibition or reduction of interaction (eg, binding) between sortilin and one or more sortilin ligands. In some embodiments, the anti-sortilin antibodies of the present invention inhibit or reduce the interaction (eg, binding) between sortilin and one or more sortilin ligands by reducing sortilin expression (eg, by reducing cell surface levels of sortilin). In some embodiments, the anti-sortilin antibodies of the present invention inhibit or reduce the interaction (e.g., binding) between sortilin and one or more sortilin ligands by at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32% , at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, according to at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57% , at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, according to at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82% , at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, according to at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or greater at saturating antibody concentration when using any in vitro ana

- 61 046146 lysis or cell culture assay described herein or known to those skilled in the art.

In some embodiments, the anti-sortilin antibody of the present invention blocks the binding of progranulin to sortilin (Sortlin) by greater than about 90% at 50 nM IgG, or greater than about 96% at 150 nM IgG, as measured by flow cytometry. In some embodiments, the anti-sortilin antibody of the present invention blocks the binding of progranulin to sortilin by about 90.74% at 50 nM IgG, as measured by flow cytometry. In some embodiments, the anti-sortilin antibody of the present invention blocks the binding of progranulin to sortilin by about 96.5% at 150 nM IgG, as measured by flow cytometry. In some embodiments, the anti-sortilin antibody of the present invention blocks the binding of progranulin to sortilin by about 96.9% at 150 nM IgG, as measured by flow cytometry.

Decreased expression of pro-inflammatory mediators.

In some embodiments, the anti-sortilin antibodies of the present invention can reduce the expression of pro-inflammatory mediators after binding to the sortilin protein expressed in the cell.

As used herein, pro-inflammatory mediators are proteins involved directly or indirectly (eg, through pro-inflammatory signaling pathways) in a mechanism that induces, activates, enhances or otherwise reduces the inflammatory response. Any method known to those skilled in the art for identifying and characterizing proinflammatory mediators can be used.

Examples of pro-inflammatory mediators include, but are not limited to, cytokines such as type I and II interferons, IL-6, IL12p70, IL12p40, IL-1e, TNF-α, IL-8, CRP, IL-20 family members, IL-33, LIF, OSM, CNTF, GM-CSF, IL-11, IL-12, IL-17, IL-18 and CRP. Additional examples of pro-inflammatory mediators include, but are not limited to, chemokines such as CXCL1, CCL2, CCL3, CCL4 and CCL5.

In some embodiments, the anti-sortilin antibodies of the present invention can reduce the functional expression and/or secretion of pro-inflammatory mediators, IL-6, IL12p70, IL12p40, IL-1e, TNF-α, CXCL1, CCL2, CCL3, CCL4 and CCL5. In some embodiments, decreased expression of proinflammatory mediators occurs in macrophages, dendritic cells, monocytes, osteoclasts, skin Langerans cells, Kupffer cells, T cells, and/or microglial cells. Decreased expression may include, but is not limited to, decreased gene expression, decreased transcriptional expression, or decreased protein expression. Any method known to those skilled in the art for determining the expression of a gene, transcript (eg, mRNA), and/or protein can be used. For example, Northern blotting can be used to determine gene expression levels of a pro-inflammatory mediator, RT-PCR can be used to determine the level of transcription of a pro-inflammatory mediator, and Western blotting can be used to determine protein levels of a pro-inflammatory mediator.

As used herein, a pro-inflammatory mediator may have reduced expression if its expression in one or more cells of a subject receiving a sortilin agent, such as the anti-sortilin agonist antibody of the present invention, is higher than the expression of the same pro-inflammatory mediator expressed in one or more cells of a corresponding subject not receiving an anti-sortilin agonist antibody. In some embodiments, the anti-sortilin antibody of the present invention may reduce the expression of a pro-inflammatory mediator in one or more cells of a subject by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70% , at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 115%, according at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, or at least 200%, for example, compared with the expression of a proinflammatory mediator in one or more cells of a corresponding subject not receiving anti-sortilin antibody. In other embodiments, the anti-sortilin antibody may reduce the expression of a pro-inflammatory mediator in one or more cells of the subject by at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold times, at least 1.9 times, at least 2.0 times, at least 2.1 times, at least 2.15 times, at least 2.2 times, at least 2.25 times, at least 2.3 times, at least 2.35 times, at least 2.4 times, at least 2.45 times, at least 2.5 times, at least 2.55 times, at least at least 3.0 times, at least 3.5 times, at least 4.0 times, at least 4.5 times, at least 5.0 times, at least 5.5 times, at least 6 ,0 times, at least 6.5 times, at least 7.0 times, at least 7.5 times, at least 8.0 times, less

- 62 046146 at least 8.5 times, at least 9.0 times, at least 9.5 times, or at least 10 times, for example, compared with the expression of a proinflammatory mediator in one or more cells of the corresponding subject, not receiving anti-sortilin antibody.

In some embodiments, an anti-sortilin antibody according to any of the above embodiments may comprise any of the features, alone or in combination, as described in Sections 1-7 below.

(1) Binding affinity of anti-sortilin antibody.

In some embodiments of any antibody provided herein, the antibody has a dissociation constant (Kd) of <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (eg 10 ^-8 M or less, eg 10 ^-8 M to 10 ^-13 M, eg 10 ^-9 M to 10 ^-13 M).

The anti-sortilin antibodies of the present invention may have nanomolar or even picomolar affinities for the target antigen (eg, human sortilin or mammalian sortilin). In some embodiments, the binding affinity of an anti-sortilin antibody of the present invention to a target antigen (eg, human or mammalian sortilin) is measured by a dissociation constant, K _D . Dissociation constants can be determined using any analytical technique, including any biochemical or biophysical methods such as fluorescence-activated cell sorting (FACS), flow cytometry, enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), biolayer interferometry (see , e.g. Octet System from ForteBio), meso scale discover technology (see e.g. MSD-SET), isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), circular dichroism (CD), analysis method stopped flow, and colorimetric or fluorescent protein melting assays; or cell binding assay. In some embodiments, the K _D for sortilin is determined at a temperature of approximately 25°C. In some embodiments, the dissociation constant (K _D ) can be measured at 4°C or at room temperature, using, for example, FACS analysis or biolayer interferometry (BioLlayer).

In some embodiments, the K _D of sortilin is determined at a temperature of approximately 4°C. In some embodiments, the K _D is determined using a monovalent antibody (eg, Fab) or a full-length antibody in monovalent form. In some embodiments, the K _D is determined using a bivalent antibody and a monomeric recombinant sortilin protein.

In some embodiments, the KD of the anti-sortilin antibodies of the present invention for human sortilin, mammalian sortilin, or both is measured using FACS as described herein (see, for example, Examples 1 and 4). In some embodiments, the K _D of the anti-sortilin antibodies of the present invention for human sortilin, mammalian sortilin, or both is measured using biolayer interferometry as described herein (see, for example, Example 4).

In some embodiments, the anti-sortilin antibody has a dissociation constant (K _D ) with human sortilin that is up to 2.5 times lower than that of an anti-sortilin antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 56, and a light chain variable region containing the sequence SEQ ID NO: 79, wherein K _D is determined by FACS (see, for example, Example 1). In some embodiments, the anti-sortilin antibody has a dissociation constant (KD) with human sortilin that ranges from about 1.10E-8 M to about 4.68E-10 M, with the _KD determined by FACS (see , for example, example 1), or from about 270 to about 2910 pM, with KD determined by biolayer interferometry (see, for example, example 4).

In some embodiments, the KD of the anti-sortilin antibody of the present invention for human sortilin, mammalian sortilin, or both may be less than 100 nM, less than 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 9 nM, less than 8 nM, less than 7 nM, less than 6 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than 0.09 nM, less than 0.08 nM, less than 0.07 nM, less than 0.06 nM, less than 0.05 nM, less than 0.04 nM, less than 0 .03 nM, less than 0.02 nM, less than 0.01 nM, less than 0.009 nM, less than 0.008 nM, less than 0.007 nM, less than 0.006 nM, less than 0.005 nM, less than 0.004 nM, less than 0.003 nM, less than 0.002 nM, less than 0.001 nM, or less than 0.001 nM.

Anti-sortilin antibody dissociation constants (KD) for human sortilin, mammalian sortilin, or both may be less than 10 nM, less than 9.5 nM, less than 9 nM, less than 8.5 nM, less than 8 nM, less than 7.5 nM, less than 7 nM, less than 6.9 nM, less than 6.8 nM, less than 6.7 nM, less than 6.6 nM, less than 6.5 nM, less than 6.4 nM, less than 6.3 nM, less than 6.2 nM, less than 6.1 nM, less than 6 nM, less than 5.5 nM, less than 5 nM, less than 4.5 nM, less than 4 nM, less than 3.5 nM, less than 3 nM, less than 2.5 nM, less than 2 nM, less than 1.5 nM, less than 1 nM, less than 0.95 nM, less than 0.9 nM, less than 0.89 nM, less than 0.88 nM, less than 0.87 nM, less than 0.86 nM, less than 0 .85 nM, less than 0.84 nM, less than 0.83 nM, less than 0.82 nM, less than 0.81 nM, less than 0.8 nM, less than 0.75 nM, less than 63 046146 0.7 nM, less than 0.65 nM, less than 0.64 nM, less than 0.63 nM, less than 0.62 nM, less than 0.61 nM, less than 0.6 nM, less than 0.55 nM, less than 0.5 nM, less than 0 .45 nM, less than 0.4 nM, less than 0.35 nM, less than 0.3 nM, less than 0.29 nM, less than 0.28 nM, less than 0.27 nM, less than 0.26 nM, less than 0.25 nM, less than 0.24 nM, less than 0.23 nM, less than 0.22 nM, less than 0.21 nM, less than 0.2 nM, less than 0.15 nM, less than 0.1 nM, less than 0.09 nM, less than 0.08 nM, less than 0.07 nM, less than 0.06 nM, less than 0.05 nM, less than 0.04 nM, less than 0.03 nM, less than 0.02 nM, less than 0.01 nM, less than 0.009 nM, less than 0.008 nM, less than 0.007 nM, less than 0.006 nM, less than 0.005 nM, less than 0.004 nM, less than 0.003 nM, less than 0.002 nM, or less than 0.001 nM.

In some embodiments, the dissociation constant (KD) of the anti-sortilin antibody is from about 0.560 nM to about 1.63 nM, for example, when determining the _KD by FACS. In some embodiments, the dissociation constant (K _D ) of the anti-sortilin antibody is from about 0.270 nM to about 2.910 nM, for example, when determining the K _D by biolayer interferometry. In some embodiments, the antibody has a dissociation constant (K _D ) with human sortilin, murine sortilin, or both that ranges from about 0.36 nM to about 0.43 nM, or less than 1.02 nM. In some embodiments, the dissociation constant is less than 1.02 nM. In some embodiments, the anti-sortilin antibody of the present invention has a dissociation constant for human sortilin of 0.560 nM or less.

In one particular embodiment, the anti-sortilin antibody of this invention has a dissociation constant for human sortilin of approximately 0.560 nM. In one specific embodiment, the anti-sortilin antibody of this invention has a dissociation constant for human sortilin of approximately 0.423 nM. In one particular embodiment, the anti-sortilin antibody of this invention has a dissociation constant for human sortilin of approximately 0.365 nM. In one particular embodiment, the anti-sortilin antibody of this invention has a dissociation constant for human sortilin of approximately 0.344 nM. In one particular embodiment, the anti-sortilin antibody of this invention has a dissociation constant for human sortilin of approximately 0.298 nM. In one specific embodiment, the anti-sortilin antibody of this invention has a dissociation constant for human sortilin of approximately 0.270 nM. In another specific embodiment, the anti-sortilin antibody of the present invention has a dissociation constant for human sortilin of approximately 0.260 nM.

In some embodiments, the anti-sortilin antibodies of the present invention have a lower dissociation constant (K _D ) for sortilin than a control anti-sortilin antibody (e.g., a control anti-sortilin antibody containing a heavy chain variable region and a light chain variable region corresponding to S- 60). In some embodiments, the anti-sortilin antibodies of the present invention have a KD for a target (e.g., human sortilin) that is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about about about about

thirty%,

45%,

60%,

75%, at at least at least at least at least at least approximately approximately approximately

35%, 50%, 65%, 80%, at at at least at least at least at least at least approximately approximately approximately approximately

40%, 55%, 70%, 85%, at least at least at least at least about 90%, at least about 95%, or at least about 99% lower than the K _D of the control antibody against target sortilin (eg, an anti-sortilin control antibody containing a heavy chain variable region and a light chain variable region corresponding to S-60). In some embodiments, the anti-sortilin antibodies of the invention have a KD for a target (e.g., human sortilin) that is at least about 1-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 12.5 times, at least about 15 times, at least about 17.5 times, at least about 20 times, at least about 22 .5 times, at least about 25 times, at least about 27.5 times, at least about 30 times, at least about 50 times, at least about 100 times, at least about 200 times, at least at least about 300 times, at least about 400 times, at least about 500 times, at least about 600 times, at least about 700 times, at least about 800 times, at least about 900 times, or at least about 900 times at least about 1000 times lower than the KD of a control anti-sortilin antibody for the target (eg, a control anti-sortilin antibody containing a heavy chain variable region and a light chain variable region corresponding to S-60).

In some embodiments, the anti-sortilin antibodies of the present invention have a KD for human sortilin that is at least 100-fold lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, the anti-sortilin antibodies of this invention

- 64 046146 have a KD for human sortilin that is at least 50 times lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, the anti-sortilin antibodies of the present invention have a KD for human sortilin that is at least 10-fold lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, the anti-sortilin antibodies of the present invention have a K _D for human sortilin that is at least 5 times lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60. In some embodiments, the anti-sortilin antibodies of the present invention have a KD for human sortilin that is at least 2-fold lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60.

In a specific embodiment, the anti-sortilin antibody of the present invention has a KD for human sortilin that is approximately 2.79 times lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S60. In another specific embodiment, the anti-sortilin antibody of the present invention has a K _D for human sortilin that is approximately 2.05 times lower than that of an anti-sortilin antibody having a heavy chain variable region and a light chain variable region corresponding to S-60.

(2) Antibody fragments.

In some embodiments of any antibodies provided herein, the antibody antibodies are an antibody fragment. Antibody fragments include, without limitation, Fab-, Fab'-, Fab'-SH-, F(ab') ₂ -, Fv- and scFv-fragments, and other fragments described below. A review of some antibody fragments is given in Hudson et al. Nat. Med. 9:129-134 (2003). An overview of scFv fragments is given, for example, in WO 93/16185; and US Patent Nos. 5,571,894 and 5,587,458. A discussion of Fab and F(ab') ₂ fragments containing salvage receptor binding epitope residues and having an increased in vivo half-life is provided in US Patent No. 5,869,046.

Diabodies are antibody fragments with two antigen-binding sites, which can be bivalent or bispecific. See for example EP404097; WO 1993/01161; Hudson et al. Nat. Med. 9:129-134 (2003). Triabodies and tetrabodies are also described in Hudson et al. Nat. Med. 9:129-134 (2003). Single domain antibodies are antibody fragments containing all or part of the heavy chain variable domain, or all or part of the light chain variable domain of the antibody. In some embodiments, the single domain antibody is a human single domain antibody (see, for example, US Pat. No. 6,248,516).

Antibody fragments can be produced by a variety of methods, including, but not limited to, proteolytic digestion of the intact antibody, as well as production by recombinant host cells (eg, E. coli or phage), as described herein.

In some embodiments, the antibody fragment is used in combination with a second anti-sortilin antibody and/or one or more antibodies that specifically bind a pathogenic protein selected from: beta-amyloid or fragments thereof, tau protein (Tau), IAPP, alphasynuclein , TDP-43, FUS protein, prion protein, PrPSc, huntingtin, calcitonin, superoxide dismutase, ataxin, Lewy bodies, atrial natriuretic factor, islet amyloid polypeptide, insulin, apolipoprotein AI, serum amyloid A, medina, prolactin, transthyretin, lysozyme, beta -2-microglobulin, gelsolin, keratoepithelin, cystatin, immunoglobulin light chain AL, S-IBM protein, non-ATG repeat-associated (RAN) translation products, dipeptide repeat (DPR) peptides, glycine-alanine repeat (GA) peptides , glycineproline repeat (GP) peptides, glycine-arginine repeat (GR) peptides, proline-alanine repeat (PA) peptides, proline-arginine repeat (PR) peptides, and any combination thereof.

(3) Chimeric and humanized antibodies.

In some embodiments of any antibody provided herein, the antibody is a chimeric antibody. Some chimeric antibodies are described, for example, in US Pat. No. 4,816,567. In one example, the chimeric antibody comprises a non-human variable region (e.g., a variable region derived from mice, rats, hamsters, rabbits, or a non-human primate such as a monkey) and the human constant region. In another example, a chimeric antibody is a class switched antibody, the class or subclass of which has been changed from the parent antibody. Chimeric antibodies include their antigen-binding fragments.

In some embodiments of any antibody provided herein, the antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity in humans while maintaining the specificity and affinity of the original non-human antibody. In some embodiments, the humanized antibody is substantially non-immunogenic in humans. In some embodiments, the humanized antibody

- 65 046146 has substantially the same affinity for the target as the antibody of the other species from which the humanized antibody was derived. See, for example, US patents No. 5530101, 5693761; 5693762; and 5,585,089. In some embodiments, amino acids of the antibody variable domain are identified that can be modified without reducing the native affinity of the antigen binding domain while reducing its immunogenicity. See, for example, US Pat. Nos. 5,766,886 and 5,869,619. Typically, a humanized antibody contains one or more variable domains in which the HVR (or portions thereof) are derived from a non-human antibody and the FR (or portions thereof) are derived from human antibody sequences. The humanized antibody will optionally also contain at least a portion of the human constant region. In some embodiments, certain FR residues of a humanized antibody are replaced with corresponding residues of a non-human antibody (eg, an antibody from which the HVR residues are derived), for example, to restore or increase the specificity or affinity of the antibody.

A review of humanized antibodies and methods for their production is given, for example, in Almagro et al. Front. Biosci. 13:161 9-1633 (2008), and further descriptions are provided, for example, in US Pat. Nos. 5821337, 7527791, 6982321 and 7087409. Human frame regions that can be used for humanization include, but are not limited to: framework regions selected from using the best fit method (see, for example, Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from a human antibody consensus sequence with a particular subset of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA 89:4285 (1992); and Presta et al., J Immunol 151:2623 (1993)); human mature (somatically mutated) frameworks or human germline frameworks (see, for example, Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions obtained from screening FR libraries (see, for example, Baca et al. J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al. J. Biol. Chem. 271:22611-22618 (1996)).

(4) Human antibodies.

In some embodiments of any antibody provided herein, the antibody is a human antibody. Human antibodies can be produced using various methods known in the art. Human antibodies are described generally in van Dijk et al. Curr. Opin. Pharmacol. 5:368-74 (2001) and Lonberg Curr. Opin. Immunol. 20:450-459 (2008).

Human antibodies can be produced by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with human variable regions in response to an antigenic stimulus. It is possible to construct mouse strains deficient in the production of mouse antibodies with large fragments of human Ig loci, with the expectation that such mice will produce human antibodies in the absence of mouse antibodies. Large fragments of human Ig can preserve a large variety of variable gene, as well as proper regulation of antibody production and expression. By exploiting murine mechanisms of antibody diversification and selection and the absence of immunological tolerance to human proteins, the human antibody repertoire produced by such murine strains can contain high-affinity fully human antibodies against any antigen of interest, including human antigens. Using hybridoma technologies, it is possible to produce and select antigen-specific human MAbs with the desired specificity. Some typical examples of methods are described in US Patent No. 5545807, EP 546073, and EP 546073. See also, for example, US Patents No. 6075181 and 6150584 describing XENOMOUSE™ technology; US Patent No. 5,770,429 describing HUMAB® technology; US Patent No. 7,041,870 describing K-M MOUSE® technology, and US Patent Application Publication No. US 2007/0061900 describing VELOCIMOUSE® technology. Human variable regions from intact antibodies generated by such animals can be further modified, for example, by combining with another human constant region.

Human antibodies can also be produced by hybridoma-based methods. Human myeloma and murine-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described (see, for example, Kozbor J. Immunol. 133:3001 (1984), and Boerner et al. J. Immunol. 147:86 (1991) ). Human antibodies generated by human B-cell hybridoma technology are also described in Li et al. Proc. Natl. Acad. Sci. USA, 1 03:3557-3562 (2006). Additional methods include those described, for example, in US Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines). Human hybridoma technology (Tmioma technology) is also described in Vollmers et al. Histology and Histopathology 20(3):927–937 (2005), and Vollmers et al. Methods and Findings in Experimental and Clinical Pharmacology 27(3):185-91 (2005). Human antibodies can also be prepared by isolating Fv clone variable domain sequences selected from human phage display libraries. Such variable domain sequences can then be combined with the desired human constant domain. Methods for selecting human antibodies from antibody libraries

- 66 046146 san below.

In some embodiments of any antibody provided herein, the antibody is a human antibody isolated by in vitro methods and/or screening of combinatorial libraries of antibodies with the desired activity or activities. Suitable examples include, but are not limited to, phage display (CAT, Morphosys, Dyax, Biosite/Medarex, Xoma, Symphogen, Alexion (formerly Proliferon), Affimed), ribosomal display (CAT), yeast-based platforms (Adimab), and the like. In certain phage display methods, the VH and VL gene repertoires are separately cloned by polymerase chain reaction (PCR) and randomly recombined into phage libraries, which can then be screened for antigen-binding phage as described by Winter et al. Ann. Rev. Immunol. 12: 433-455 (1994). For example, various methods are known to those skilled in the art for preparing phage display libraries and screening such libraries for antibodies having desired binding characteristics. See also Sidhu et al. J. Mol. Biol. 338(2):299-310, 2004; Lee et al. J. Mol. Biol. 340(5):1073-1093, 2004; Fellows Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004); and Lee et al. J. Immunol. Methods 284(-2):1 19-132 (2004). Phage typically displays antibody fragments, as single chain Fv (scFc) fragments or as Fab fragments. Libraries from immunized sources produce high-affinity antibodies to the immunogen without the need for hybridoma construction. Alternatively, the naive repertoire (eg, human) can be cloned to provide a single source of antibodies to a wide range of foreign as well as self antigens, without any immunization, as described by Griffiths et al. EMBO J. 12: 725-734 (1993). Finally, naive libraries can also be generated artificially by cloning unrearranged V gene segments from stem cells, and using PCR primers containing random sequences to encode the highly variable regions of HVR3 and to perform in vitro rearrangement, as described by Hoogenboom et al. J. Mol. Biol., 227:381-388, 1992. Patent publications describing human antibody phage libraries include, for example, US Patent No. 5,750,373, and US Patent Publications No. 2007/0292936 and 2009/0002360. Antibodies isolated from human antibody libraries are considered herein to be human antibodies or human antibody fragments.

(5) Constant regions containing Fc regions.

In some embodiments of any antibody provided herein, the antibody comprises an Fc. In some embodiments, the Fc is of the human IgG1, IgG2, IgG3, and/or IgG4 isotype. In some embodiments, the antibody is of the IgG class, the IgM class, or the IgA class.

In some embodiments of any antibody provided herein, the antibody is of the IgG2 isotype. In some embodiments, the antibody comprises a human IgG2 constant region. In some embodiments, the human IgG2 constant region comprises an Fc region. In some embodiments, the antibody induces one or more sortilin activities (or) independently of binding to an Fc receptor. In some embodiments, the antibody binds an inhibitory Fc receptor. In some embodiments, the inhibitory Fc receptor is an inhibitory Fc receptor gamma IIB (FcyIIB).

In some embodiments of any antibody provided herein, the antibody is of the IgG1 isotype. In some embodiments, the antibody comprises a mouse IgG1 constant region. In some embodiments, the antibody comprises a human IgG1 constant region. In some embodiments, the human IgG1 constant region comprises an Fc region. In some embodiments, the antibody binds an inhibitory Fc receptor. In some embodiments, the inhibitory Fc receptor is an inhibitory Fc gamma receptor IIB (FcyIIB).

In some embodiments of any antibody provided herein, the antibody is of the IgG4 isotype. In some embodiments, the antibody comprises a human IgG4 constant region. In some embodiments, the human IgG4 constant region comprises an Fc region. In some embodiments, the antibody binds an inhibitory Fc receptor. In some embodiments, the inhibitory Fc receptor is an inhibitory Fc receptor gamma IIB (FcyIIB).

In some embodiments of any antibody provided herein, the antibody is an IgG2/4 hybrid isotype. In some embodiments, the antibody comprises an amino acid sequence comprising human IgG2 amino acids 118-260 according to EU numbering and human IgG4 amino acids 261-447 according to EU numbering (WO 1997/11971; WO 2007/106585).

In some embodiments, the Fc region increases clustering without complement activation compared to a corresponding antibody containing an Fc region that does not contain amino acid substitutions. In some embodiments, the antibody induces one or more activities of a target specifically bound by the antibody. In some embodiments, the antibody binds to sortilin.

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It may also be desirable to modify the anti-sortilin antibody of the present invention to modify effector function and/or to increase the serum half-life of the antibody. For example, the Fc receptor binding site in the constant region can be modified or mutated to eliminate or reduce binding affinity to certain Fc receptors, such as FcyRI, FcyRII, and/or FcyIIIB to reduce antibody-dependent cell-mediated cytotoxicity. In some embodiments, effector function is disrupted by removing N-glycosylation of the Fc region (eg, in the CH2 domain of an IgG) of the antibody. In some embodiments, effector function is disrupted by modifying regions such as 233236, 297 and/or 327-331 of human IgG, as described in WO 99/58572 and Armor et al. Molecular Immunology 40:585-593 (2003); Reddy et al. J Immunology 164:1925–1933 (2000). In other embodiments, it may also be desirable to modify the anti-sortilin antibody of the present invention to modify effector function to increase detection selectivity towards ITIM-containing FcgRIIb (CD32b) to enhance clustering of sortilin antibodies on adjacent cells without activating humoral responses, including antibody-dependent cell-mediated cytotoxicity and antibody-dependent cellular phagocytosis.

To increase the half-life of serum antibodies, a salvage receptor binding epitope (especially an antibody fragment) can be included in the antibody, as described, for example, in US Pat. No. 5,739,277. As used herein, the term salvage receptor binding epitope refers to an Fc region epitope IgG molecule (eg, IgGi, IgG2, IgG3, or IgG ₄ ) responsible for increasing the in vivo half-life of the IgG molecule from serum. Other amino acid sequence modifications.

(6) Multispecific antibodies.

Multispecific antibodies are those that have binding specificity for at least two different epitopes, including those located on the same or different polypeptides (eg, one or more sortilin polypeptides of this invention). In some embodiments, the multispecific antibody may be a bispecific antibody. In some embodiments, the multispecific antibody may be a trispecific antibody. In some embodiments, the multispecific antibody may be a tetraspecific antibody. Such antibodies can be obtained from full-length antibodies or antibody fragments (eg, F(ab') ₂ bispecific antibodies). In some embodiments, the multispecific antibody comprises a first antigen binding region that binds to a first sortilin site and contains a second antigen binding region that binds to a second sortilin site. In some embodiments, the multispecific antibodies comprise a first antigen binding region that binds sortilin and a second antigen binding region that binds a second polypeptide.

Provided herein are multispecific antibodies comprising a first antigen binding region, the first antigen binding region comprising six HVRs of an antibody described herein that binds sortilin, and a second antigen binding region that binds a second polypeptide. In some embodiments, the first antigen binding region comprises a VH or VL of an antibody described herein.

In some embodiments of any of the multispecific antibodies, the second polypeptide is (a) an antigen that facilitates transport across the blood-brain barrier; (b) an antigen that facilitates transport across the blood-brain barrier selected from transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and 2) , diphtheria toxin receptor, CRM197, llama single domain antibody, TMEM 30(A), protein transduction domain, TAT, Syn-B, penetratin, polyarginine peptide, angiopep peptide and ANG1005; (c) a pathogenic protein selected from beta-amyloid, oligomeric beta-amyloid, beta-amyloid plaques, amyloid precursor protein or fragments thereof, tau protein (Tau), IAPP, alpha-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open reading frame 72), c9RAN protein, prion protein, PrPSc, huntingtin, calcitonin, superoxide dismutase, ataxin, ataxin 1, ataxin 2, ataxin 3, ataxin 7, ataxin 8, ataxin 10, Lewy bodies, atrial natriuretic factor , islet amyloid polypeptide, insulin, apolipoprotein AI, serum amyloid A, medina, prolactin, transthyretin, lysozyme, beta-2-microglobulin, gelsolin, keratoepithelin, cystatin, immunoglobulin light chain AL, S-IBM protein, non-ATG repeat associated (RAN) translation products, peptides with dipeptide repeats (DPR), peptides with glycine-alanine repeats (GA), peptides with glycine-proline repeats (GP), peptides with glycine-arginine repeats (GR), peptides with proline-alanine repeats (PA ), ubiquitin, and proline-arginine repeat (PR) peptides; (d) ligands and/or proteins expressed on immune cells, wherein the ligands and/or proteins are selected from CD40, OX40, ICOS, CD28, CD137/4-1BB, CD27, GITR, PD-L1, CTLA-4, PD -L2, PD-1, B7-H3, B7-H4, HVEM, BTLA, KIR, GAL9, TIM3, A2AR, LAG-3, and phosphatidylserine; and/or (e) a protein, lipid, polysaccharide or glycolipid expressed on one or more tumor cells, and any combination thereof.

- 68 046146

Those skilled in the art are aware of numerous antigens that facilitate transport across the blood-brain barrier (see, for example, Gabathuler R. Neurobiol. Dis. 37:48-57 (2010)). Such second antigens include, but are not limited to, transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, including CRM197 (a non-toxic mutant of diphtheria toxin), single domain llama antibodies such as TMEM 30(A) (flippase), protein transduction domains such as TAT, Syn-B, or penetratin, polyarginine or generally positively charged peptides, angiopep peptides such like ANG1005 (see, e.g., Gabathuler, 2010), and other cell surface proteins present in increased abundance on endothelial cells of the blood-brain barrier (see, e.g., Daneman et al. PLoS One 5(10):e13741 (2010)) .

Polyvalent antibodies can recognize sortilin antigen, as well as, without limitation, additional antigens - Ae peptide antigen, or α-synuclein protein antigen, or tau protein antigen, or TDP-43 protein antigen, or prion protein antigen, or huntingtin protein antigen, or RAN translation product antigen, including dipeptide repeats (DPR peptides) consisting of glycine-alanine (GA), glycine-proline (GP), glycine-arginine (GR), proline-alanine (PA), or proline-arginine (PR) , insulin receptor, insulin-like growth factor receptor. Transferrin receptor or any other antigen that facilitates the movement of an antibody across the blood-brain barrier. In some embodiments, the second polypeptide is transferrin. In some embodiments, the second polypeptide is tau protein (Tau). In some embodiments, the second polypeptide is Ae (amyloid beta peptide). In some embodiments, the second polypeptide is TREM2. In some embodiments, the second polypeptide is α-synuclein.

A multivalent antibody contains at least one polypeptide chain (preferably two polypeptide chains), wherein the polypeptide chain or chains contain two or more variable domains. For example, the polypeptide chain or chains may comprise VD1-(X1) _n -VD2-(X2) _n -Fc, where VD1 is a first variable domain, VD2 is a second variable domain, Fc is one Fc region polypeptide chain, X1 and X2 is an amino acid or a polypeptide, and n is 0 or 1. Likewise, the polypeptide chain or chains may comprise a V||-C chain.'ii 1-flexible linker-V _H -C _H 1-Fc region; or chain V _H -C _H 1-V _H -C _H 1-Fc region. The multivalent antibody herein preferably further comprises at least two (and preferably four) light chain variable domain polypeptides. A multivalent antibody herein may, for example, contain from about two to about eight light chain variable domain polypeptides. The light chain variable domain polypeptides provided herein contain a light chain variable domain and optionally further contain a CL domain.

Methods for producing multispecific antibodies include, but are not limited to, recombinant coexpression of two immunoglobulin heavy chain-light chain pairs having different specificities (see Milstein and Cuello Nature 305:537 (1983), WO 93/08829, and Traunecker et al. EMBO J. 10:3655 (1991)), and knob-in-hole technology (see, for example, US Pat. No. 5,731,168). See also WO 2013/026833 (CrossMab). Multispecific antibodies can also be produced by electrostatic interaction targeting technology to produce Fc heterodimeric antibody molecules (WO 2009/089004A1); by cross-linking two or more antibodies (see, for example, US patent No. 4676980); using leucine; using diabody technology to produce bispecific antibody fragments (see, for example, Hollinger et al. Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993)); and by using single chain Fv (scFv) dimers (see, eg, Gruber et al. J. Immunol. 152:5368 (1994)); and by producing trispecific antibodies as described, for example, in Tutt et al. J. Immunol. 147:60 (1991).

Recombinant antibodies with three or more functional antigen binding sites, including Octopus antibodies (octopus antibodies with three or more antigen binding sites and two or more variable regions), are also included herein (see, for example, US 2006/0025576). The antibody herein also includes a Dual Acting FAb or DAF (Dual Acting FAb) containing an antigen binding site that binds to multiple sortilins (see, for example, US 2008/0069820).

(7) Antibodies with improved stability.

Modifications to the amino acid sequence of the anti-sortilin antibodies of the present invention, or fragments of such antibodies, are also provided to improve stability during manufacture, storage, and in vivo administration. For example, it may be desirable to reduce the degradation of antibodies or antibody fragments of the present invention in various ways, including, but not limited to, oxidation and deamidation. Variants of antibody amino acid sequences or antibody fragments are produced by introducing appropriate nucleotide changes into the nucleic acid encoding the antibodies or antibody fragments, or by peptide synthesis. Such modifications include

- 69 046146 include, for example, deletions and/or insertions and/or substitutions of residues in the amino acid sequences of the antibody. Any combination of deletions, insertions and substitutions can be made to produce a finished construct, provided that the finished construct has the desired characteristics (ie, reduced sensitivity to degradation).

In some embodiments, the asparagine site (N33) in the HVR-L1 region of the anti-sortilin antibody of the present invention may be susceptible to degradation by deamidation. In some embodiments, the asparagine site (N33) in the HVR-L1 region of S-60-15 (SEQ ID NO: 8) may be susceptible to deamidation. After deamidation, the asparagine site (N33) in the HVR-L1 region of S-60-15 results in the replacement of Asn with Asp/u30-Asp (IsoAsp). In some embodiments, the asparagine site (N33) in the HVR-L1 region of S-60-15 may be replaced to prevent or reduce deamidation. Non-limiting examples of S-60-15 amino acid sequence variants having amino acid substitutions at the asparagine site (N33) of the HVR-L1 region include S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G ], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60 -15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S -60-15.15 [N33A], S-60-15.16 [N33M] or S-6015.17 [N33L].

(8) Antibody variants.

In some embodiments of any of the antibodies provided herein, variants of the amino acid sequences of the antibodies are provided. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody.

(i) Variants of substitutions, insertions and deletions.

In some embodiments of any of the antibodies provided herein, antibody variants are provided having one or more amino acid substitutions. Variants of antibody amino acid sequences can be obtained by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and/or insertions and/or substitutions of residues in the amino acid sequences of the antibody.

Table 37

Amino acid substitutions

Original balance Typical examples of substitutions Preferred Substitutions Ala (A) Vai; Leu; He Vai Arg(R) Lys; Gin; Asn Lys Asn(N) Gin; His; Asp, Lys; Arg Gin Asp (D) Glu; Asn Glu Cys(C) Ser; Ala Ser Gln(Q) Asn; Glu Asn Glu (E) Asp; Gin Asp Gly (G) Ala Ala His(H) Asn; Gin; Lys; Arg Arg He(I) Leu; Vai; Met; Ala; Ph; norleucine Leu Leu (L) norleucine; He; Vai; Met; Ala; Phe He Lys (K) Arg; Gin; Asn Arg Met(M) Leu; Ph; He Leu Phe (F) Leu; Vai; He; Ala; Tyr Tyr Pro (P) Ala Ala Ser(S) Thr Thr Thr(T) Ser Ser Trp(W) Tyr; Phe Tyr Tyr(Y) Trp; Ph; Thr; Ser Phe Vai (V) He; Leu; Met; Ph; Ala; norleucine Leu

Significant modifications of the biological properties of an antibody are made by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone

- 70 046146 at the site of substitution, for example, with sheet or helix conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the volume of the side chain. Natural residues are divided into groups based on the general properties of their side chains:

(1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

(2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

(3) acidic: Asp, Glu;

(4) basic: His, Lys, Arg;

(5) residues affecting chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

For example, non-conservative substitutions may involve exchanging any member of one of these classes for a member from another class. Such substituted residues may be introduced, for example, into regions of a human antibody that are homologous to non-human antibodies, or into non-homologous regions of the molecule.

When introducing changes to a polypeptide or antibody described herein, in accordance with certain embodiments, the amino acid hydropathic index may be taken into account. Each amino acid was assigned a hydropathic index based on its hydrophobicity and charge characteristics. They have the following values: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).

Experts understand the importance of the hydropathicity index of amino acids in imparting a biological interaction function to a protein. Kyte et al. J. Mol. Biol., 157:105-131 (1982). It is known that certain amino acids can be replaced by other amino acids that have a similar index or quantification of hydropathicity, and still retain similar biological activity. When making changes based on the hydropathic index, some embodiments include substitution of amino acids having hydropathic indices that differ by no more than ±2. Some embodiments include a difference of no more than ±1, and some embodiments include a difference of no more than ±0.5.

Those skilled in the art will also appreciate that similar amino acid substitutions can be effectively made on the basis of hydrophilicity, particularly when the resulting biologically functional protein or peptide is intended for use in immunological embodiments, as is the case here. In some embodiments, the highest local average hydrophilicity of a protein, determined by the hydrophilicity of its adjacent amino acids, correlates with its immunogenicity and antigenicity, i.e. with the biological properties of protein.

The amino acid residues were assigned the following hydrophilicity values: arginine (+3.0); lysine (+3.0±1); aspartate (+3.0±1); glutamate (+3.0±1); series (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5±1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5) and tryptophan (-3.4). When making changes based on similar hydrophilicity values, some embodiments involve replacing amino acids having hydrophilicity values that differ by no more than ±2, some embodiments include values that differ by no more than ±1, and some embodiments include values that differ no more than ±0.5. Based on hydrophilicity, epitopes of primary amino acid sequences can also be identified. Such regions are also called epitope core regions.

In some embodiments, substitutions, insertions, or deletions may be located in one or more HVRs, as long as such changes do not significantly reduce the ability of the antibody to bind antigen. For example, conservative changes (eg, conservative substitutions as contemplated herein) that do not significantly reduce binding affinity can be made in HVR. Such changes may, for example, be located outside of the antigen-contacting residues in the HVR. In some embodiments of the VH and VL sequence variants noted above, each HVR is unchanged, or contains no more than one, two, or three amino acid substitutions.

Amino acid sequence insertions include amino- and/or carboxy-terminal fusions ranging in length from one residue to polypeptides containing one hundred or more residues, as well as intrachain insertions of one or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue. Other insertional variants of antibody molecules include fusion of the antibody at the N- or C-terminus with an enzyme (eg, for ADEPT (antibody-mediated enzyme and prodrug therapy)) or polypeptide that increases the half-life of serum antibodies.

Any cysteine residue not involved in maintaining the proper conformation of the antibody can also be replaced, usually with serine, to increase the oxidative stability of the molecule.

- 71 046146 ly and prevent aberrant cross-linking. Conversely, cysteine linkage(s) may be added to an antibody to increase its stability (particularly in cases where the antibody is an antibody fragment, such as an Fv fragment).

(ii) Glycosylation variants.

In some embodiments of any antibody provided herein, the antibody is modified to increase or decrease the degree of glycosylation of the antibody. The addition or deletion of glycosylation sites in an antibody can conveniently be accomplished by altering the amino acid sequence so as to create or remove one or more glycosylation sites.

Glycosylation of antibodies is usually N-linked or O-linked. N-linking refers to the attachment of a carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are recognition sequences for the enzymatic attachment of a carbohydrate moiety to the asparagine side chain. Thus, the presence of any of these tripeptide sequences in a polypeptide creates a potential glycosylation site. Linked glycosylation refers to the addition of one of the sugars N-acetylgalactosamine, galactose or xylose, to an hydroxy amino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine can also be used.

The addition of glycosylation sites to an antibody is conveniently accomplished by altering the amino acid sequence so that it contains one or more of the tripeptide sequences described above (for N-linked glycosylation sites). Changes can also be made by adding or replacing one or more serine or threonine residues to the original antibody sequence (for O-linked glycosylation sites).

In cases where the antibody contains an Fc region, the carbohydrate attached to it may be modified. Native antibodies produced by mammalian cells typically contain a branched biantennary oligosaccharide, typically N-linked to Asn297 in accordance with the Kabat numbering of the CH2 domain of the Fc region. The oligosaccharide may include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to GlcNAc in the stem of the biantennary oligosaccharide structure. In some embodiments, modifications to the oligosaccharide in an antibody of the invention may be made to create variants of the antibody with certain improved properties.

In one embodiment, antibody variants are provided having a carbohydrate structure lacking fucose attached (directly or indirectly) to the Fc region. See, for example, US Patent Publications No. 2003/0157108 and 2004/0093621. Examples of publications regarding defucosylated or fucose-deficient antibody variants include: US 2003/0157108; US 2003/0115614; US 2002/0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004). Examples of cell lines capable of producing defucosylated antibodies include CHO Led 3 cells deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US 2003/0157108) and knockout cell lines such as alpha-1,6-fucosyltransferase gene knockouts, FUT8, CHO cells (see, for example, Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004) and Kanda et al. Biotechnol. Bioeng. 94 ( 4):680-688 (2006)).

(iii) Modified constant regions.

In some embodiments of any antibodies provided herein, the Fc antibodies are isotypes and/or modifications of the Fc antibodies. In some embodiments, the Fc isotype and/or modification of the antibody is capable of binding to the Fc gamma receptor.

In some embodiments of any antibody provided herein, the modified Fc antibody is an IgG1 modified Fc. In some embodiments, the modified IgG1 Fc contains one or more modifications. For example, in some embodiments, the modified IgG1 Fc contains one or more amino acid substitutions (eg, compared to a wild-type Fc region of the same isotype). In some embodiments, one or more amino acid substitutions are selected from N297A (Bolt S et al. (1993) Eur J Immunol 23:403-411), D265A (Shields et al. (2001) R. J. Biol. Chem. 276, 6591-6604 ), L234A, L235A (Hutchins et al. (1995) Proc Natl Acad Sci USA, 92:11980-11984; Alegre et al., (1994) Transplantation 57:1537-1543. 31; Xu et al., (2000) Cell Immunol, 200:16-26), G237A (Alegre et al. (1994) Transplantation 57:1537-1543. 31; Xu et al. (2000) Cell Immunol, 200:16-26), C226S, C229S, E233P , L234V, L234F, L235E (McEarchern et al., (2007) Blood, 109:1185-1192), P331S (Sazinsky et al., (2008) Proc Natl Acad Sci USA 2008, 105:20167-20172), S267E, L328F, A330L, M252Y, S254T, and/or T256E, where the amino acid positions follow the EU numbering scheme. In some embodiments of any antibody provided herein, the antibody is of the IgG1 isotype and the Fc region contains amino acid substitutions at positions L234A, L235A, and P331S, wherein the residue position numbering follows the EU numbering system.

- 72 046146

In some embodiments, any of the modified IgG1 Fcs, the Fc contains the N297A mutation in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, contain mutations D265A and N297A in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc contains the D270A mutation in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains the L234A and L235A mutations in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, comprise the L234A and G237A mutations in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, comprise the L234A, L235A, and G237A mutations in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc, contains one or more (including all) of the mutations P238D, L328E, E233, G237D, H268D, P271G, and A330R in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc contains one or more of the S267E/L328F mutations according to the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc contains mutations P238D, L328E, E233D, G237D, H268D, P271G, and A330R in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, comprise mutations P238D, L328E, G237D, H268D, P271G, and A330R in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc contains mutations P238D, S267E, L328E, E233D, G237D, H268D, P271G, and A330R in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc contains mutations P238D, S267E, L328E, G237D, H268D, P271G, and A330R in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, comprise mutations C226S, C229S, E233P, L234V, and L235A in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, comprise the L234F, L235E, and P331S mutations in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fcs, comprise the S267E and L328F mutations in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc, contains the S267E mutation in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the Fc comprises replacing the heavy chain constant region 1 (CH1) and hinge region of IgG1 with CH1 and the hinge region of IgG2 (amino acids 118-230 of IgG2 according to the EU numbering system) with a kappa light chain.

In some embodiments, any of the modified IgG1 Fcs, the Fc, contains two or more amino acid substitutions that increase clustering of the antibody without activating complement compared to a corresponding antibody having an Fc region that does not contain two or more amino acid substitutions. Accordingly, in some embodiments of any of the modified Fc IgG1, the modified Fc IgG1 is an antibody comprising an Fc region, wherein the antibody contains an amino acid substitution at position E430G and one or more amino acid substitutions in the Fc region at a residue position selected from: L234F, L235A, L235E, S267E, K322A, L328F, A330S, P331S, and any combination thereof, in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G, L243A, L235A, and P331S in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G and P331S in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G and K322A in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G, A330S, and P331S in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G, K322A, A330S, and P331S in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G, K322A, and A330S in accordance with the EU numbering system. In some embodiments, the modified IgG1 Fc contains amino acid substitutions at positions E430G, K322A, and P331S in accordance with the EU numbering system.

In some embodiments, any of the modified IgG1 Fcs, the modified IgG1 Fc may further comprise and be combined with the A330L mutation (Lazar et al. Proc Natl Acad Sci USA, 103:4005-4010 (2006)), or one or more L234F, L235E mutations and/or P331S (Sazinsky et al. Proc Natl Acad Sci USA, 105:20167–20172 (2008)), according to the EU numbering system, to eliminate complement activation. In some embodiments, any of the modified IgG1 Fcs, the modified IgG1 Fcs may further comprise one or more of A330L, A330S, L234F, L235E, and/or P331S in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 Fcs, the modified IgG1 Fcs may further comprise one or more mutations to increase the half-life of the human serum antibody (e.g., one or more (including all) of the mutations M252Y, S254T, and T256E according to the numbering system EU). In some embodiments, any of the modified IgG1 Fcs, modified

- 73 046146 citated IgG1 Fc may further comprise one or more of E430G, E430S, E430F, E430T, E345K, E345Q, E345R, E345Y, S440Y, and/or S440W according to the EU numbering system.

Other aspects of the present invention relate to antibodies having modified constant regions (ie Fc regions). An antibody dependent on FcgR binding to activate target receptors may lose its agonist activity as a result of modification to eliminate FcgR binding (see, e.g., Wilson et al. Cancer Cell 19:101-113 (2011); Armor at al Immunology 40:585-593 (2003); and White et al. Cancer Cell 27:138-148 (2015). As such, it is believed that the anti-sortilin antibody of the present invention with appropriate epitope specificity can activate a target antigen with minimal undesirable effects when the antibody has a human IgG2 isotype Fc domain (CH1 and hinge region) or another type of Fc domain capable of preferentially binding inhibitory FcgRIIB receptors, or its variant.

In some embodiments of any antibody provided herein, the modified Fc antibody is a modified Fc IgG2. In some embodiments, the modified IgG2 Fc contains one or more modifications. For example, in some embodiments, the modified IgG2 Fc contains one or more amino acid substitutions (eg, compared to a wild-type Fc region of the same isotype). In some embodiments, any of the modified IgG2 Fcs, one or more amino acid substitutions are selected from V234A (Alegre et al. Transplantation 57:1537-1543 (1994); Xu et al. Cell Immunol, 200:16-26 (2000)); G237A (Cole et al. Transplantation, 68:563-571 (1999)); H268Q, V309L, A330S, P331S (US 2007/0148167; Armour et al. Eur J Immunol 29: 2613-2624 (1999); Armour et al. The Hematology Journal l (Suppl. 1):27 (2000); Armour et al. al. The Hematology Journal 1(Suppl.1):27 (2000)), C219S, and/or C220S (White et al. Cancer Cell 27, 138-148 (2015)); S267E, L328F (Chu et al. Mol Immunol, 45:3926-3933 (2008)); and M252Y, S254T, and/or T256E, according to the EU numbering system. In some embodiments, any of the modified IgG2 Fcs, the Fc contains amino acid substitutions at positions V234A and G237A in accordance with the EU numbering system. In some embodiments, any of the modified IgG2 Fcs, the Fc, contains amino acid substitutions at positions C219S or C220S in accordance with the EU numbering system. In some embodiments, any of the modified IgG2 Fcs, the Fc, contains amino acid substitutions at positions A330S and P331S in accordance with the EU numbering system. In some embodiments, any of the modified IgG2 Fcs, the Fc contains amino acid substitutions at positions S267E and L328F in accordance with the EU numbering system.

In some embodiments, any of the modified IgG2 Fcs, the Fc comprises a C127S amino acid substitution in accordance with the EU numbering system (White et al., (2015) Cancer Cell 27, 138148; Lightle et al. Protein Sci. 19:753-762 (2010 ); and WO 2008/079246). In some embodiments, any of the modified IgG2 Fcs, the antibody has an IgG2 isotype with a kappa light chain constant domain that contains the C214S amino acid substitution in accordance with the EU numbering system (White et al. Cancer Cell 27:138-148 (2015); Lightle et al Protein Sci 19:753-762 (2010); and WO 2008/079246).

In some embodiments, any of the modified IgG2 Fcs, the Fc comprises a C220S amino acid substitution in accordance with the EU numbering system. In some embodiments of any of the modified IgG2 Fcs, the antibody is an IgG2 isotype with a kappa light chain constant domain that contains the C214S amino acid substitution in accordance with the EU numbering system.

In some embodiments, any of the modified IgG2 Fcs, the Fc comprises a C219S amino acid substitution in accordance with the EU numbering system. In some embodiments of any of the modified IgG2 Fcs, the antibody is of an IgG2 isotype with a kappa light chain constant domain that contains the C214S amino acid substitution in accordance with the EU numbering system.

In some embodiments, any of the modified IgG2 Fcs, the Fc comprises an IgG2 isotype heavy chain constant domain 1 (CH1) and a hinge region (White et al. Cancer Cell 27:138-148 (2015)). In some embodiments, any of the modified IgG2 Fc, CH1 of the IgG2 isotype, and the hinge region comprise amino acid sequence 118-230 according to the EU numbering system. In some embodiments, any of the modified IgG2 Fcs, the Fc region of the antibody comprises amino acid substitution S267E, amino acid substitution L328F, or both, and/or amino acid substitution N297A or N297Q in accordance with the EU numbering system.

In some embodiments, any of the modified IgG2 Fcs, the Fc further comprises one or more amino acid substitutions at positions E430G, E430S, E430F, E430T, E345K, E345Q, E345R, E345Y, S440Y, and S440W in accordance with the EU numbering system. In some embodiments, any of the modified IgG2 Fcs may further comprise one or more mutations to increase the half-life of the human serum antibody (e.g., one or more (including all) of the mutations M252Y, S254T, and T256E according to the EU numbering system) . In some embodiments, any of the modified IgG2 Fcs, the Fc may further comprise A330S and P331S.

In some embodiments, any of the modified IgG2 Fcs, the Fc is a hybrid IgG2/4 Fc. In some embodiments, the IgG2/4 Fc hybrid comprises amino acids

- 74 046146 (aa) 118-260 IgG2 and amino acids 261-447 IgG4. In some embodiments, any of the modified IgG2 Fcs, the Fc, contains one or more amino acid substitutions at positions H268Q, V309L, A330S, and P331S in accordance with the EU numbering system.

In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains one or more additional amino acid substitutions selected from A330L, L234F; L235E or P331S, according to the EU numbering system; and any combination thereof.

In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains one or more amino acid substitutions at a residue position selected from C127S, L234A, L234F, L235A, L235E, S267E, K322A, L328F, A330S, P331S, E345R, E430G , S440Y, and any combination thereof, in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G, L243A, L235A, and P331S in accordance with the EU numbering system. In some embodiments, either of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G and P331S in accordance with the EU numbering system. In some embodiments, either of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G and K322A in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G, A330S, and P331S in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G, K322A, A330S, and P331S in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G, K322A, and A330S in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E430G, K322A, and P331S in accordance with the EU numbering system. In some embodiments, either of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions S267E and L328F in accordance with the EU numbering system. In some embodiments, either of the modified IgG1 and/or IgG2 Fcs, the Fc contains an amino acid substitution at position C127S in accordance with the EU numbering system. In some embodiments, any of the modified IgG1 and/or IgG2 Fcs, the Fc contains amino acid substitutions at positions E345R, E430G, and S440Y in accordance with the EU numbering system.

In some embodiments of any antibody provided herein, the modified Fc antibody is a modified Fc IgG4. In some embodiments, the modified IgG4 Fc contains one or more modifications. For example, in some embodiments, the modified IgG4 Fc contains one or more amino acid substitutions (eg, compared to a wild-type Fc region of the same isotype). In some embodiments, any of the modified IgG4 Fc, one or more amino acid substitutions are selected from L235A, G237A, S229P, L236E (Reddy et al. J Immunol 164:1925-1933(2000)), S267E, E318A, L328F, M252Y, S254T , and/or T256E according to the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc may further comprise L235A, G237A, and E318A in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc may further comprise S228P and L235E in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the modified IgG4 Fc may further comprise S267E and L328F in accordance with the EU numbering system.

In some embodiments, any of the modified IgG4 Fcs, the modified IgG4 Fc contains and may be combined with the S228P mutation according to the EU numbering system (Angal et al. Mol Immunol. 30:105-108 (1993)) and/or one or more mutations described in (Peters et al. J Biol Chem. 287(29):24525-33 (2012)) to enhance antibody stabilization.

In some embodiments, any of the modified IgG4 Fcs, the modified IgG4 Fc may further comprise one or more mutations to increase the half-life of the human serum antibody (e.g., one or more (including all) of the mutations M252Y, S254T, and T256E according to the numbering system EU).

In some embodiments, any of the modified IgG4 Fcs, the Fc comprises L235E in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc contains one or more amino acid substitutions at a residue position selected from C127S, F234A, L235A, L235E, S267E, K322A, L328F, E345R, E430G, S440Y, and any combination thereof, according to with EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc, contains amino acid substitutions at positions E430G, L243A, L235A, and P331S in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc, contains amino acid substitutions at positions E430G and P331S in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc, contains amino acid substitutions at positions E430G and K322A in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc comprises an amino acid

- 75 046146 menu in position E430 in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc region contains amino acid substitutions at positions E430G and K322A in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc, contains amino acid substitutions at positions S267E and L328F in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc contains an amino acid substitution at position C127S in accordance with the EU numbering system. In some embodiments, any of the modified IgG4 Fcs, the Fc, contains amino acid substitutions at positions E345R, E430G, and S440Y in accordance with the EU numbering system.

(9) Other modifications of the antibody.

In some embodiments of any of the antibodies, the antibody is a derivative. The term derivative refers to a molecule that contains a chemical modification other than an insertion, deletion, or substitution of amino acids (or nucleic acids). In some embodiments, the derivative contains covalent modifications, including, without limitation, chemical bonding to polymers, lipids, or other organic or inorganic moieties. In some embodiments, a chemically modified antigen binding protein may have a longer half-life in circulation than an antigen binding protein that has not been chemically modified. In some embodiments, the chemically modified antigen binding protein may have an improved ability to target desired cells, tissues, and/or organs. In some embodiments, the derivatized antigen binding protein is covalently modified to include one or more attached water-soluble polymer moieties, including, without limitation, polyethylene glycol, polyoxyethylene glycol, or polypropylene glycol. See, for example, US Pat. Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417, 4,791,192, and 4,179,337. In some embodiments, the derivatized antigen binding protein comprises one or more polymers, including, but not limited to, monomethoxy polyethylene glycol, dextran, cellulose, ethylene glycol/propylene glycol copolymers Kolya, carboxymethylcellulose, polyvinylpyrrolidone, poly1,3-dioxolane, poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyamino acids (homopolymers or random copolymers), poly(vinylpyrrolidone)polyethylene glycol, propylene glycol homopolymers, polypropylene oxide/ethylene oxide copolymer, polyoxyethylated polyols (eg glycerol) and polyvinyl alcohol, as well as mixtures of such polymers.

In some embodiments, the derivative is covalently modified with polyethylene glycol (PEG) subunits. In some embodiments, one or more water-soluble polymers are added at one or more specific positions, for example, at the amino terminus of the derivative. In some embodiments, one or more water-soluble polymers are randomly attached to one or more side chains of the derivative. In some embodiments, PEG is used to improve the therapeutic properties of an antigen binding protein. In some embodiments, PEG is used to enhance the therapeutic properties of the humanized antibody. Some such methods are described, for example, in US Pat. No. 6,133,426, which is hereby incorporated by reference for all purposes.

Peptide analogues are widely used in the pharmaceutical industry as non-peptide drugs with properties similar to the matrix peptide. These types of non-peptide compounds are called peptide mimetics or peptidomimetics. Fauchere, J. Adv. Drug Res., 15:29 (1986); and Evans et al. J. Med. Chem., 30:1229 (1987), which are incorporated herein by reference for all purposes. Such compounds are often developed using computer molecular modeling. Peptidomimetics, which are structurally similar to therapeutically useful peptides, can be used to produce similar therapeutic or prophylactic effects. In general, peptidomimetics are structurally similar to a reference polypeptide (i.e., a polypeptide having biochemical properties or pharmacological activity), such as a human antibody, but optionally have one or more peptide bonds replaced by a bond selected from: -CH2NH-, -CH2S-, -CH2-CH2-, -CH=cH- (cis- and trans-), -COCH2-, -CH(OH)CH ₂ -, and CH2SO-, using methods well known to those skilled in the art. Systematic replacement of one or more amino acids of the consensus sequence with a D-amino acid of the same type (eg, D-lysine instead of L-lysine) can be used in some embodiments to produce more stable peptides. In addition, conformationally constrained peptides containing a consensus sequence or a substantially identical variant of the consensus sequence can be prepared by methods known in the art (Rizo and Gierasch Ann. Rev. Biochem., 61:387 (1992), incorporated herein by reference for any purpose); for example, by adding internal cysteine residues capable of forming intramolecular disulfide bridges that cyclize the peptide.

Drug conjugation involves combining a biologically active cytotoxic (anti-cancer) payload or drug with an antibody that specifically targets a specific tumor marker (eg, a polypeptide that is ideally present only in or on tumor cells). Antibodies find these proteins in the body and attach

- 76 046146 to the surface of cancer cells. A biochemical reaction between the antibody and the target protein (antigen) initiates a signal in the tumor cell, which then takes up or internalizes the antibody along with the cytotoxin. Once the antibody-drug conjugate (ADC) is internalized, the cytotoxic drug is released and kills the cancer. Thanks to this targeting, the drug ideally has lower side effects and creates a wider therapeutic window than other chemotherapy agents. The technical basis of antibody conjugation is disclosed and known to those skilled in the art (see, for example, Jane de Lartigue, OncLive July 5, 2012; ADC Review on antibody-drug conjugates; and Ducry et al. Bioconjugate Chemistry 21 (1):5-13 (2010) .

Binding and other assays.

The anti-sortilin antibodies of the present invention can be tested for antigen binding activity, for example, by known methods such as ELISA, surface plasmon resonance (SPR), Western blotting, flow cytometry, FACS, biolayer interferometry, etc.

In some embodiments, competition assays can be used to identify an antibody that competes with any antibody described herein. In some embodiments, competition assays can be used to identify an antibody that competes with any of the antibodies listed in Table. 1-30, or contains a heavy chain variable region and a light chain variable region of an antibody selected from S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60 -7, S-60-8, S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)] , S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60- 15.6 [N33H], S-6015.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S- 60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L ], S-60-16, S-60-18, S-60-19, and S-60-24, for binding to sortilin. In some embodiments, such a competing antibody binds to the same epitope (eg, a linear or conformational epitope) as binding to any of the antibodies that are listed in Table. 1-30, or contain a heavy chain variable region and a light chain variable region of an antibody selected from S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60 -7, S-60-8, S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)] , S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S60-15.6 [ N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S- 60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L ], S-60-16, S-60-18, S-60-19, and S-60-24. A detailed description of typical methods for mapping the epitope to which an antibody binds is given in Morris (1996) Epitope Mapping Protocols, in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ).

In a typical example of a competition assay, immobilized sortilin or cells expressing sortilin on the cell surface are incubated in a solution containing a first labeled antibody that binds sortilin (eg, human or non-human primate) and a second unlabeled antibody that is tested. on its ability to compete with the first antibody for binding to sortilin. The second antibody may be present in the supernatant of the hybridoma. As a control, immobilized sortilin or cells expressing sortilin are incubated in a solution containing the first labeled antibody but not the second unlabeled antibody. After incubation under conditions permissive for binding of the first antibody to sortilin, excess unbound antibody is removed and the amount of label bound to immobilized sortilin or cells expressing sortilin is measured. If the amount of label bound to immobilized sortilin or cells expressing sortilin is significantly reduced in the test sample compared to the control sample, then this indicates that the second antibody is competing with the first antibody for binding to sortilin. See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

Ligand binding assays.

This document further provides screening methods for anti-sortilin antibodies that bind His131, Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg38 2, Tyr386, Ser595, and/ or Glu700 human sortilin (SEQ ID NO: 81); or mammalian sortilin amino acid residues that correspond to one or more amino acid residues of His131, Val132, Pro133, Leu 134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223 , Ser227, Ser242, Lys243, Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg 382, Tyr386, Ser595, and /or Glu700 SEQ ID NO: 81, thereby blocking interactions between sortilin and sortilin ligand (eg, progranulin, proneurotrophin, pro-NGF, pro-BDNF, pro-OT3, p75, APP, LpL, APOA5, APOE). In some embodiments, a peptide library can be synthesized in which the sortilin protein is resolved into successive 15-mer and 25-mer peptides separated by a single amino acid residue and then spot applied to filters. Ligand binding testing can then be carried out.

- 77 046146 on to determine its ability to interact with the receptor peptide or with human sortilin library peptides that have been mutated, for example, at positions His131, Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203 , Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260, Ser3O5, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe3 50, Tyr351, Ser352, Ile353 , Asn373, Ser379, Arg382, Tyr386, Ser595, and/or Glu700, in the presence or absence of anti-sortilin antibodies by spot binding assay (SPOT) (eg, Frank, R and Overwin, H (1996) Methods. Mol. Biol. 66, 149-169; Reineke, U et al., (2002) J. Immunol. Methods 267, 13-26; and Andersen, OS et al., (2010) J. Biological Chemistry 285, 1221012222).

Further provided herein are screening methods for anti-sortilin antibodies that block the interactions (eg, binding) of sortilin and a sortilin ligand (eg, progranulin, proneurotrophin, pro-NGF, pro-BDNF, προ-ΝΤ3, p75, APP, LpL, APOA5 , APOE). In some embodiments, the interactions between sortilin and sortilin ligands (e.g., progranulin, proneurotrophin, pro-NGF, pro-BDNF, npo-NT3, p75, APP, LpL, APOA5, APOE) can be characterized using surface plasmon resonance analysis (e.g. Skeldal, S et al., (2012) J Biol Chem., 287:43798; and Andersen, OS et al., (2010) The Journal Of Biological Chemistry, 285,12210-12222, pulldown analysis (e.g. , Andersen, OS et al., (2010) The Journal Of Biological Chemistry, 285,12210-12222, cellulose-bound proteins (e.g., Andersen, OS et al., (2010) The Journal Of Biological Chemistry, 285,12210- 12222), proximity ligation assays (eg, Gustafsen, C et al., (2013) The Journal of Neuroscience, 33:64-71), and/or alkaline phosphatase-labeled ligands in cellular binding assays (eg , Hu, F et al., (2010) Neuron 68, 654-667).

Cellular assays.

Further provided herein are screening methods for a sortilin binding antagonist, such as an anti-sortilin antibody, which involve contacting an agent (eg, an anti-sortilin antibody) with a cell expressing the sortilin protein on the cell surface. In some embodiments, the agent and the cell are further contacted with a sortilin ligand of the invention. In some embodiments, the cell itself expresses the sortilin ligand of the present invention. Cellular methods are particularly useful for screening and validating sortilin binding antagonists (eg, anti-sortilin antibodies) by assessing the effect on the interaction between sortilin and sortilin ligand in the context of a cell.

Accordingly, certain aspects of the present invention relate to a cell expressing the sortilin protein of the present invention on its cell surface. In some embodiments, the cell endogenously expresses the sortilin protein of the present invention. In some embodiments, the cell is recombinantly modified to express the sortilin protein of the invention. In any of these embodiments, the sortilin protein of this invention (endogenous or recombinant) encoded by the polynucleotide will preferably contain at least protein domains necessary for post-translational processing, membrane translocation, and cell surface targeting, including, but not limited to, signaling peptide and transmembrane domain. In some embodiments, the signal peptide and/or transmembrane domain may refer to the signal peptide and/or transmembrane domain of endogenous sortilin. In other embodiments, the signal peptide and/or transmembrane domain may refer to an exogenous signal peptide and/or transmembrane domain known to enhance cell surface expression in the desired host cell. In preferred embodiments, the sortilin protein will also contain a domain sufficient to bind the sortilin ligand of the present invention.

In such embodiments, any cell that expresses the sortilin protein of the present invention on its cell surface can be used. In some embodiments, the cell endogenously expresses the sortilin protein of the invention on its cell surface. In some embodiments, a cell is recombinantly modified to express the sortilin protein of the invention on its cell surface. Any suitable sortilin ligand of the present invention can be used such that it retains the ability to bind to sortilin protein expressed on the cell surface. The sortilin ligand does not necessarily need to be fluorescently labeled. Levels of sortilin ligand can be detected using any assay known in the art, including, but not limited to, ELISA, SPR (surface plasmon resonance), Western blotting, mass spectrometry, immunoprecipitation, peptide microarrays, etc.

In some embodiments, the methods disclosed herein include culturing a cell that expresses both the sortilin protein on its cell surface and the sortilin ligand in the medium, under conditions under which the sortilin protein and the sortilin ligand are expressed and the sortilin ligand is released into the medium ; contacting the cell with an agent (eg, an anti-sortilin antibody) under conditions under which the sortilin protein is able to bind to the sortilin ligand;

- 78 046146 and detecting an increase in the level of sortilin ligand in the medium, compared to the level of sortilin ligand in the medium in the absence of the specified agent. An increase in the level of sortilin ligand indicates that the agent is an antagonist of sortilin binding. Without wishing to be bound by theory, it is believed that the interaction between the cell surface expressed protein sortilin and the secreted sortilin ligand will result in endocytosis and lysosomal degradation of the sortilin ligand. Therefore, it is believed that reducing this interaction (eg, by adding the sortilin binding antagonist of the present invention) results in an increase in the level of sortilin ligand in the medium over time.

In such embodiments, any cell that expresses the sortilin protein of the present invention on its cell surface and expresses and secretes the sortilin ligand of the present invention can be used. In some embodiments, a cell may endogenously express the sortilin protein of the invention on its cell surface. In some embodiments, a cell may endogenously express and secrete a sortilin ligand of the present invention. In some embodiments, the cell is a U-251 cell and the sortilin ligand is a progranulin protein. In some embodiments, a cell can be recombinantly modified to express the sortilin protein of the invention on its cell surface. In some embodiments, a cell can be recombinantly modified to express and secrete a sortilin ligand of the present invention.

In any of the cellular assays described herein, the sortilin ligand of this invention can be used. In some embodiments, the sortilin ligand is a progranulin protein. The sortilin ligand may be a full-length protein, or may be a sortilin-binding peptide fragment thereof.

Nucleic acids, vectors and host cells.

The anti-sortilin antibodies of the present invention can be produced using recombinant methods and compositions, for example, as described in US Pat. No. 4,816,567. In some embodiments, isolated nucleic acids are provided containing a nucleotide sequence encoding any of the anti-sortilin antibodies of the present invention. Such nucleic acids may encode an amino acid sequence containing VL and/or an amino acid sequence containing VH of anti-sortilin antibodies (eg, light and/or heavy chains of the antibody). In some embodiments, one or more vectors (eg, expression vectors) containing such nucleic acids are provided. In some embodiments, a host cell containing such a nucleic acid is also provided. In some embodiments, the host cell contains (e.g., has been transduced): (1) a vector containing a nucleic acid that encodes an amino acid sequence containing a VL antibody and an amino acid sequence containing a VH antibody, or (2) a first vector containing a nucleic acid that encodes an amino acid sequence containing a VL antibody, and a second vector containing a nucleic acid that encodes an amino acid sequence containing a _VH antibody. In some embodiments, the host cell is a eukaryotic, such as a Chinese hamster ovary (CHO) cell, or a lymphoid cell (eg, a YO, NSO, Sp20 cell). Host cells of this invention also include, but are not limited to, isolated cells, in vitro cultured cells, and ex vivo cultured cells.

Methods for producing the anti-sortilin antibody of the present invention are provided. In some embodiments, the method includes culturing a host cell of the present invention containing a nucleic acid encoding an anti-sortilin antibody under conditions suitable for expressing the antibody. In some embodiments, the antibody is then isolated from the host cell (or host cell culture medium).

To recombinantly produce the anti-sortilin antibody of the present invention, a nucleic acid encoding the anti-sortilin antibody is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell. Such nucleic acid can be readily isolated and sequenced using conventional procedures (eg, by using oligonucleotide probes capable of specifically binding to genes encoding antibody heavy and light chains).

Suitable vectors containing a nucleic acid sequence encoding any of the anti-sortilin antibody polypeptides of the present invention, or cell surface expressed fragments or polypeptides thereof (including antibodies) described herein include, but are not limited to, cloning vectors and expression vectors. Suitable cloning vectors can be constructed according to standard techniques, or can be selected from a large number of available cloning vectors. Although the cloning vector chosen may vary depending on the host cell to be used, suitable cloning vectors generally have the ability to self-replicate, may have a single target for a particular restriction endonuclease, and/or may carry genes for a marker that can be

- 79 046146 used in the selection of clones containing the vector. Suitable examples include plasmids and bacterial viruses, for example pUC18, pUC19, Bluescript (eg pBS SK+) and its derivatives, mp18, mp19, pBR322, pMB9, ColE1, pCR1, RP4, phage DNA, and shuttle vectors such as pSA3 and pAT28. These and many other cloning vectors are available from commercial suppliers such as BioRad, Strategene, and Invitrogen.

Suitable host cells for cloning or expressing antibody-encoding vectors include prokaryotic or eukaryotic cells. For example, the anti-sortilin antibodies of the present invention can be produced in bacteria, particularly if glycosylation and Fc effector function are not required. For the expression of antibody fragments and polypeptides in bacteria (eg, US patents No. 5648237, 5789199 and 5840523). Once expressed, the antibody can be isolated from the bacterial cell paste in a soluble fraction and can be further purified.

In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeast are also suitable hosts for the cloning or expression of antibody-encoding vectors, including fungal and yeast strains whose glycosylation pathways have been humanized to produce an antibody with a partially or fully human glycosylation pattern ( for example, Gerngross Nat. Biotech. 22:1409-1414 (2004); and Li et al. Nat. Biotech. 24:210-215 (2006)).

Suitable host cells for expression of the glycosylated antibody can also be obtained from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculovirus strains have been identified that can be used in combination with insect cells, particularly for transfection of Spodoptera frugiperda (fall armyworm) cells. Plant cell cultures can also be used as hosts (eg, US Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 describing PLANTIBODIES™ antibody production technology in transgenic plants).

Vertebrate cells can also be used as hosts. For example, mammalian cell lines adapted for suspension growth may be used. Other examples of useful mammalian host cell lines include the monkey kidney line CV1 transformed with SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, for example, in Graham et al. J. Gen Virol. 36:59 (1977)); baby hamster kidney (BHK) cells; mouse Sertoli cells (TM4 cells, as described, for example, in Mather, Biol. Reprod. 23:243251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma (HELA) cells; canine kidney cells (MDCK); buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); murine mammary tumor (MMT 060562); TRI cells, as described, for example, in Mather et al. Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al. Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0 and Sp2/0. A review of some mammalian host cell lines suitable for antibody production is given, for example, in Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

Pharmaceutical compositions.

Provided herein are pharmaceutical compositions and/or pharmaceutical formulations comprising the anti-sortilin antibodies of the present invention and a pharmaceutically acceptable carrier.

In some embodiments, the pharmaceutically acceptable carrier is preferably non-toxic to recipients at the dosages and concentrations used. The antibodies described herein can be formulated in solid, semi-solid, liquid or gaseous form. Examples of such formulations include, but are not limited to, tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalations, gels, microspheres and aerosols. Pharmaceutically acceptable carriers may include, depending on the desired formulation, pharmaceutically acceptable non-toxic carriers or diluents, which are substances commonly used to formulate pharmaceutical compositions for administration to animals or humans. In some embodiments, the pharmaceutical composition may include auxiliary materials to modify, maintain or maintain, for example, the pH value, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption or permeability of the composition.

In some embodiments, pharmaceutically acceptable carriers include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine); antimicrobial agents; antioxidants (such as ascorbic acid, sodium sulfite or sodium bisulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates or other organic acids); excipients (such as mannitol or glycine); chelating agents (such as ethylenediaminetetra

- 80 046146 acetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose or dextrins); proteins (such as serum albumin, gelatin or immunoglobulins); coloring agents, flavoring agents and diluents; emulsifiers; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as Pluronics, PEGs, sorbitan esters, polysorbates such as polysorbate 20, polysorbate 80, Triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancers (such as sucrose or sorbitol); tonicity agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol, sorbitol); delivery means; thinners; excipients and/or pharmaceutical adjuvants. Additional examples of compositions suitable for various types of administration are provided in Remington: The Science and Practice of Pharmacy, Pharmaceutical Press 22nd ed. (2013). A brief review of drug delivery methods is given in Langer, Science 249:1527-1533 (1990).

Compositions suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injectable solutions, which may contain antioxidants, buffers, bacteriostats and solutes to render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions, which may contain suspending agents, solubilizers, thickeners, stabilizers and preservatives.

Sosots can be optimized for retention and stabilization in the brain or central nervous system. In cases where an agent is administered to a cranial compartment, it is desirable that the agent is retained in the compartment and does not diffuse or otherwise pass through the blood-brain barrier. Stabilization techniques include cross-linking, multimerization, or coupling to groups such as polyethylene glycol, polyacrylamide, neutral protein carriers, etc., to increase molecular weight.

Other strategies to increase retention include incorporating an antibody, such as the anti-sortilin antibody of the present invention, into a biodegradable or bioerodible implant. The rate of release of the therapeutically active agent is controlled by the rate of transport through the polymer matrix, and the biodegradation of the implant. Implants may be particles, sheets, patches, plaques, fibers, microcapsules, etc., and may be of any size or shape compatible with the chosen insertion site. Biodegradable polymer compositions suitable for use may be organic esters or ethers which, when degraded, provide physiologically acceptable degradation products, including monomers. Anhydrides, amides, orthoesters, etc., alone or in combination with other monomers, may be used. The polymers will be condensation polymers. Polymers can be cross-linked or non-cross-linked. Of particular interest are polymers of hydroxyaliphatic carboxylic acids, whether homo- or copolymers, and polysaccharides. Polyesters of interest include polymers of D-lactic acid, L-lactic acid, racemic lactic acid, glycolic acid, polycaprolactone, and combinations thereof. Polysaccharides of interest are calcium alginate and functionalized celluloses, in particular carboxymethylcellulose esters, characterized by water insolubility, with a molecular weight of from about 5 kDa to 500 kDa, etc. Biodegradable hydrogels can also be used in the implants of this invention. Hydrogels are typically a copolymer material characterized by the ability to absorb liquid.

Pharmaceutical dosages.

The antibody provided herein (and any additional therapeutic agent) may be administered by any suitable means, including parenteral, intrapulmonary, intranasal, intralesional, intracerebrospinal, intracranial, intraspinal, intrasynovial, intrathecal, oral, topical, or inhalation. Parenteral infusions include intramuscular, intravenous bolus or continuous infusion over a period of time, intra-arterial, intra-articular, intraperitoneal or subcutaneous administration. In some embodiments, administration is intravenous administration. In some embodiments, administration is subcutaneous. Dosing may be administered by any suitable route, for example by injection, such as intravenous or subcutaneous injection, depending in part on whether the administration is short-term or chronic. Various dosage regimens are contemplated herein, including, without limitation, single or multiple dosing at different times, bolus dosing, and pulse infusion.

The antibody compositions provided herein will be formulated, dosed and

- 81 046146 be administered in accordance with the principles of good medical practice. Factors requiring consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the route of administration, the schedule of administration, and other factors known to the practitioner. The antibody composition need not necessarily be formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of antibody present in the composition, the type of disorder or treatment, and other factors noted above. They are generally used at the same dosages and routes of administration described herein, or in amounts from about 1 to 99% of the dosages described herein, or at any dose and route of administration that are empirically/ clinically determined to be suitable.

Dosages for a particular anti-sortilin antibody may be determined empirically for individuals receiving one or more administrations of the anti-sortilin antibody. Individuals are administered increasing doses of anti-sortilin antibody. To assess the effectiveness of the anti-sortilin antibody, the clinical symptom of any diseases, disorders or conditions of this invention (eg, frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, traumatic brain injury, spinal cord injury, long-term depression) can be monitored , atherosclerotic vascular diseases, and undesirable symptoms of normal aging).

For the prevention or treatment of a disease, a suitable dosage of an antibody of this invention (when used alone or in combination with one or more other additional therapeutic agents) will depend on the type of disease requiring treatment, the type of antibody, the severity and course of the disease, whether the antibody is administered whether for prophylactic or therapeutic purposes, previous therapy, the patient's medical history and response to the antibody, and the opinion of the treating physician. The antibody may be administered to the patient at one time or over a number of treatment sessions.

Depending on the type and severity of the disease, about 1 mcg/kg to 15 mg/kg (eg, 0.1 mg/kg-10 mg/kg) of antibody may be an initial trial dosage to administer to the patient, e.g., through one or more individual administration sessions, or by continuous infusion. One typical daily dose may range from about 1 mcg/kg to 100 mg/kg or more, depending on the factors noted above. For repeated administration over several days or a longer period of time, depending on the condition, treatment will generally be continued until the desired suppression of disease symptoms is achieved. One typical dose of antibody will range from about 0.05 mg/kg to about 10 mg/kg. Thus, the patient may be administered one or more doses of about 0.5 mg/kg, 2.0 mg/kg, 4.0 mg/kg, or 10 mg/kg (or any combination thereof). Such doses may be administered periodically, for example, once a week or every three weeks (eg, such that the patient receives from about two to about twenty or, for example, about six doses of the antibody). In some embodiments, the dosing frequency is three times a day, twice a day, once a day, every two days, once a week, every two weeks, every four weeks, every five weeks, every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every ten weeks, or once a month, once every two months, once every three months, or less often. A high initial loading dose may be administered, followed by one or more lower doses. However, other administration schedules may be useful. The progress of such therapy is easily monitored by conventional methods and tests.

Therapeutic use.

In certain aspects, the anti-sortilin antibodies of this invention can be used to prevent, reduce the risk of, or treat an individual with a disease, disorder, or lesion. The anti-sortilin antibodies of the present invention can be used to prevent, reduce the risk of, or treat cell death (eg, neuronal cell death), frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, traumatic brain injury, spinal cord injury, long-term depression, atherosclerotic vascular diseases, undesirable symptoms of normal aging, dementia, mixed dementia, Creutzfeldt-Jakob disease, normal pressure hydrocephalus, amyotrophic lateral sclerosis, Huntington's disease, tauopathy, stroke, acute trauma, chronic trauma, lupus, acute and chronic colitis, disease Crohn's, inflammatory bowel disease, ulcerative colitis, malaria, essential tremor, central nervous system lupus, Behçet's disease, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, intervertebral disc degeneration, Shy-Drager syndrome, progressive supranuclear palsy, corticobasal ganglion degeneration , acute disseminated encephalomyelitis, granulomatous disorders, sarcoidosis, diseases of aging, age-related macular degeneration, glaucoma, retinitis pigmentosa, retinal degeneration, respiratory tract infection, sepsis, eye infection, systemic infection, inflammatory disorders, arthritis, multiple sclerosis, metabolic

- 82 046146 deterioration, obesity, insulin resistance, type 2 diabetes, tissue or vascular damage, trauma, and/or one or more undesirable symptoms of normal aging.

In certain aspects, this document provides a method of preventing, reducing the risk of, or treating an individual with a disease, disorder, or lesion, comprising administering to the individual in need thereof a therapeutically effective amount of an anti-sortilin antibody of the present invention. In some embodiments, the disease, disorder, or lesion is selected from the group consisting of frontotemporal dementia, progressive supranuclear palsy, Alzheimer's disease, vascular dementia, seizure disorder, retinal dystrophy, amyotrophic lateral sclerosis, traumatic brain injury, spinal cord injury, dementia, stroke, Parkinson's disease, acute disseminated encephalomyelitis, retinal degeneration, age-related macular degeneration, glaucoma, multiple sclerosis, septic shock, bacterial infection, arthritis and osteoarthritis.

Certain aspects of the present invention provide methods for increasing progranulin levels in an individual in need thereof, such as in the brain, blood, and/or peripheral organs of the individual, by administering to the individual a therapeutically effective amount of one or more anti-sortilin antibodies of the present invention. Other aspects of the present invention provide methods for increasing extracellular levels of progranulin by contacting one or more cells with one or more anti-sortilin antibodies of the present invention. In some embodiments, progranulin levels are increased without decreasing cellular sortilin levels. Other aspects of the present invention provide methods for reducing cellular levels of sortilin in an individual in need thereof, such as in the brain and/or peripheral organs of the individual, by administering to the individual a therapeutically effective amount of one or more anti-sortilin antibodies of the present invention. Other aspects of the present invention provide methods for reducing cellular sortilin levels of one or more cells, comprising contacting one or more cells with one or more anti-sortilin antibodies of the present invention.

Additional aspects of the present invention provide methods for increasing the effective concentrations of progranulin and/or decreasing the effective concentrations of the neurotrophin of the present invention (e.g., proneurotrophin-3, proneurotrophin-4/5, proneurotrophins, pro-NGF, proBDNF, neurotrophin-3, neurotrophin-4/5 , NGF, BDNF, etc.), neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5) , apolipoprotein E (APOE), PCSK9, and receptor associated protein (RAP) to an individual in need thereof by administering to the individual a therapeutically effective amount of an anti-sortilin antibody of the invention to inhibit the interaction between sortilin and progranulin, a neurotrophin of the invention (e.g., proneurotrophins, proneurotrophin -3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, sortilin propeptide (Sort-pro), protein -amyloid precursor (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), and/or receptor associated protein (RAP).

The present invention also provides methods for inhibiting the interaction between sortilin and progranulin, a neurotrophin of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF , BDNF, etc.), neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and/or receptor-associated protein (RAP); as well as one or more activities of the sortilin, progranulin, neurotrophin of this invention (for example, proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF etc.), neurotensin, p75, sortilin propeptide (Sortpro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE) , and/or receptor-associated protein (RAP) in an individual by administering to the individual a therapeutically effective amount of an anti-sortilin antibody of the present invention.

In certain aspects, this disclosure provides a method of preventing, reducing the risk of, or treating an individual with a disease, disorder, or lesion, comprising administering to the individual in need thereof a therapeutically effective amount of an anti-sortilin antibody of the present invention.

As disclosed herein, the anti-sortilin antibodies of this invention can be used to prevent, reduce the risk of, or treat frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, traumatic brain injury, spinal cord injury, long-term depression , atherosclerotic vascular diseases, undesirable symptoms of normal aging, dementia, mixed dementia

- 83 046146 tion, Creutzfeldt-Jakob disease, normal pressure hydrocephalus, amyotrophic lateral sclerosis, Huntington's disease, tauopathy, stroke, acute injury, chronic injury, lupus, acute and chronic colitis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, malaria, essential tremor, lupus of the central nervous system, Behçet's disease, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, intervertebral disc degeneration, Shy-Drager syndrome, progressive supranuclear palsy, corticobasal ganglion degeneration, acute disseminated encephalomyelitis, granulomatous disorders, sarcoidosis, diseases of aging , age-related macular degeneration, glaucoma, retinitis pigmentosa, retinal degeneration, respiratory tract infection, sepsis, eye infection, systemic infection, inflammatory disorders, arthritis, multiple sclerosis, metabolic disorder, obesity, insulin resistance, type 2 diabetes, tissue or vascular damage, trauma , and one or more undesirable symptoms of normal aging.

In some embodiments, the invention provides methods for preventing, reducing the risk of, or treating an individual with frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, traumatic brain injury, spinal cord injury, long-term depression, atherosclerotic vascular disease, unwanted symptoms of normal aging, dementia, mixed dementia, Creutzfeldt-Jakob disease, normal pressure hydrocephalus, amyotrophic lateral sclerosis, Huntington's disease, tauopathy, stroke, acute trauma, chronic trauma, lupus, acute and chronic colitis, Crohn's disease, inflammatory bowel disease, nonspecific ulcerative colitis, malaria, essential tremor, central nervous system lupus, Behçet's disease, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, intervertebral disc degeneration, Shy-Drager syndrome, progressive supranuclear palsy, corticobasal ganglion degeneration, acute disseminated encephalomyelitis, granulomatous disorders , sarcoidosis, diseases of aging, age-related macular degeneration, glaucoma, retinitis pigmentosa, retinal degeneration, respiratory tract infection, sepsis, eye infection, systemic infection, inflammatory disorders, arthritis, multiple sclerosis, metabolic disorder, obesity, insulin resistance, type 2 diabetes, damage tissue or blood vessels, trauma, and one or more undesirable symptoms of normal aging, in an individual in need thereof, by administering to the individual a therapeutically effective amount of an anti-sortilin antibody of the present invention to: (i) inhibit the interaction between sortilin and progranulin, a neurotrophin of the present invention (e.g. proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, sortilin propeptide ( Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), and/or receptor associated protein (RAP); and/or (ii) inhibiting one or more of the activities of the sortilin, progranulin, neurotrophin of the present invention (e.g. proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/ 5, NGF, BDNF, etc.), neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), beta-amyloid (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), and/or receptor-associated protein (RAP). In some embodiments, the present invention provides methods for inducing wound healing in an individual in need thereof by administering to the individual a therapeutically effective amount of an anti-sortilin antibody of the present invention.

The present invention also provides methods for promoting cell survival, such as the survival of neuronal cells, by administering an anti-sortilin antibody of the present invention to inhibit the interaction between sortilin and progranulin, a neurotrophin of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, amyloid precursor protein (APP), and amyloid beta (A beta) peptide. An anti-sortilin antibody can be introduced into cells in vitro to promote cell survival. Alternatively, an anti-sortilin antibody may be administered in vivo (eg, by administering the antibody to an individual) to promote cell survival.

In certain aspects, this disclosure provides a method of inhibiting one or more of neuroinflammation, axonopathy characterized by short axonal sprouting and aberrant branching, microglial activation, and inflammatory response, comprising administering to a subject a therapeutically effective amount of an anti-sortilin antibody of the present invention.

This invention provides methods for inhibiting neuroinflammation, axonopathy characterized by short axonal sprouting and aberrant branching, microglial activation, and the inflammatory response, and enhancing wound healing, autophagy, and clearance of aggregates.

- 84 046146 proteins by administering an anti-sortilin antibody of the present invention to inhibit the interaction between sortilin and progranulin, a neurotrophin of the present invention (e.g. proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3 , neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, amyloid precursor protein (APP), and amyloid beta (A beta) peptide. Antibody against sortilin can be introduced into cells in vitro. Alternatively, the anti-sortilin antibody can be administered in vivo (eg, by administering the antibody to an individual).

In certain aspects, this disclosure provides a method of enhancing one or more of wound healing, autophagy, and the clearance of aggregated proteins, comprising administering to an individual a therapeutically effective amount of an anti-sortilin antibody of the invention.

In certain aspects, this disclosure provides a method of preventing, reducing the risk of, or treating an individual with arthritis, comprising administering to the individual a therapeutically effective amount of an anti-sortilin antibody of the present invention.

The present invention also provides methods for reducing the expression of one or more pro-inflammatory mediators by administering to an individual in need thereof an anti-sortilin antibody of the present invention. In some embodiments, the one or more proinflammatory mediators are selected from IL-6, IL12p70, IL12p40, IL-1e, TNF-α, CXCL1, CCL2, CCL3, CCL4, and CCL5.

In some embodiments, the method of this invention includes an anti-sortilin antibody comprising two or more anti-sortilin antibodies.

Dementia.

Dementia is a nonspecific syndrome (i.e., a set of signs and symptoms) manifesting in a previously unaffected individual as a severe loss of general cognitive ability beyond what is expected with normal aging. Dementia can be static, resulting from a unique global brain injury. Alternatively, dementia may be progressive, resulting in long-term deterioration due to physical impairment or illness. Although dementia is significantly more common in the geriatric population, it can also occur in people under 65 years of age. Cognitive areas affected in dementia include, but are not limited to, memory, attention span, language, and problem solving. Typically, symptoms must have been present for at least six months before an individual is diagnosed with dementia.

Common examples of forms of dementia include, but are not limited to, frontotemporal dementia, Alzheimer's disease, vascular dementia, semantic dementia, and dementia with Lewy bodies.

Without wishing to be bound by theory, it is believed that administration of the anti-sortilin antibody of the present invention can prevent, reduce the risk of, and/or treat dementia. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with dementia (e.g., neurotrophic and/or neuronal survival activity, and anti-inflammatory activity.

Frontotemporal dementia.

Frontotemporal dementia (FTD) is a condition resulting from progressive degeneration of the frontal lobe of the brain. Over time, degeneration may involve the temporal lobe. Second only to Alzheimer's disease (AD) in prevalence, FTD accounts for 20% of cases of presenile dementia. Clinical features of FTD include memory deficits, behavioral abnormalities, personality changes, and language impairments (Cruts, M. & Van Broeckhoven, C., Trends Genet. 24:186-194 (2008); Neary, D., et al., Neurology 51 :1546-1554 (1998); Ratnavalli, E., Brayne, C., Dawson, K. & Hodges, J. R., Neurology 58:1615-1621 (2002)).

A significant proportion of cases of FTD are inherited in an autosomal dominant manner, but even within a single family, symptoms can span the spectrum from FTD with behavioral disturbances to primary progressive aphasia to corticobasal ganglion degeneration. FTD, like most neurodegenerative diseases, can be characterized by the abnormal presence of specific protein aggregates in the affected brain. Historically, early descriptions of FTD noted the presence of intraneuronal accumulations of hyperphosphorylated tau protein in neurofibrillary tangles or Pick bodies. The causative role of microtubule-associated protein tau has been confirmed by the identification of mutations in the gene encoding tau protein in several families (Hutton, M., et al., Nature 393:702-705 (1998). However, in the brain of most people, affected by FTD, there was no accumulation of hyperphosphorylated tau protein, and immunoreactivity for ubiquitin (Ub) and TAR DNA binding protein (TDP43) was demonstrated (Neumann, M., et al., Arch. Neurol. 64:1388-1394 (2007) Most of these cases of FTD with Ub inclusions (FTD-U) have been shown to carry mutations in the progranulin gene.

Progranulin mutations lead to haploinsufficiency and are known to be present in nearly 50% of cases of familial FTD, making progranulin mutation an important genetic factor favoring FTD. Without wishing to be bound by theory, we point out that it is believed that heterozygous

- 85 046146 the loss-of-function nature of progranulin mutations suggests that in healthy individuals, progranulin expression plays a dose-dependent critical role in protecting healthy individuals from the development of FTD. Accordingly, increasing progranulin levels by inhibiting the interaction between sortilin and progranulin may prevent, reduce the risk of, and/or treat FTD.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat FTD. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with FTD.

Alzheimer's disease.

Alzheimer's disease (AD) is the most common form of dementia. There is no cure for the disease, which worsens as it progresses and gradually leads to death. AD is most often diagnosed in people over the age of 65. However, a less common form of Alzheimer's disease with early onset may appear much earlier.

Common symptoms of Alzheimer's disease include behavioral symptoms such as difficulty remembering recent events; cognitive symptoms, confusion, irritability and aggression, emotional lability, language problems, and long-term memory loss. As the disease progresses, bodily functions are lost, ultimately leading to death. Alzheimer's disease develops over an unknown and variable period of time before it becomes fully apparent, and can progress while remaining undiagnosed for years.

Sortilin has been shown to bind to amyloid precursor protein (APP) and the APP processing enzyme BACE1. Without wishing to be bound by theory, these interactions are believed to be involved in Alzheimer's disease. Accordingly, and without wishing to be limited by theory, it is believed that the anti-sortilin antibodies of this invention can be used to inhibit such interactions, and prevent, reduce the risk of, or treat Alzheimer's disease in individuals in need thereof.

In some embodiments, and without wishing to be bound by theory, anti-sortilin antibodies of the invention that inhibit the interaction between sortilin and the neurotrophins of the invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), p75, amyloid precursor protein (APP), and/or amyloid beta (A beta) peptide, or which inhibit one or multiple activities of sortilin may be used to prevent, reduce the risk of, or treat Alzheimer's disease in individuals in need thereof.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat Alzheimer's disease. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with Alzheimer's disease.

Vascular dementia.

Vascular dementia (VaD) is a subtle, progressive decline in memory and other cognitive functions thought to be caused by cerebrovascular disease (vascular disease in the brain). Cerebrovascular disease is a progressive change in our blood vessels (vasculature) in the brain (cerebrum). The most common vascular change associated with age is the accumulation of cholesterol and other substances in the walls of blood vessels. This leads to thickening and hardening of the walls, as well as narrowing of blood vessels, which can lead to a decrease or even complete cessation of blood flow to the areas of the brain supplied by the affected artery. Patients with vascular dementia often exhibit symptoms similar to those observed in patients with Alzheimer's disease (AD). However, the associated changes in the brain are not caused by the pathology of Alzheimer's disease, but by chronically reduced blood flow in the brain, gradually leading to dementia. Vascular dementia is considered one of the most common types of dementia in older people. Symptoms of vascular dementia include difficulty remembering, difficulty organizing and solving complex problems, slowed thinking, distraction or lack of concentration, difficulty recalling words, changes in mood or behavior such as depression, irritability or apathy, and hallucinations or delusions.

Without wishing to be bound by theory, one or more activities of sortilin, or one or more interactions between sortilin and progranulin, are believed to be the neurotrophins of this invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, lipoprotein lipase, apolipoprotein AV, and/or receptor-associated protein are involved in vascular dementia. Accordingly, and without wishing to be limited by theory, it is believed that anti-sortilin antibodies of the present invention that inhibit the interaction between sortilin and the neurotrophins of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF , neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.),

- 86 046146 neurotensin, p75, sortilin propeptide (Sort-pro), amyloid precursor protein (APP), amyloid beta (A beta) peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), and /or receptor-associated protein (RAP); or that inhibit one or more of the activities of sortilin, may be used to prevent, reduce the risk of, or treat vascular dementia in individuals in need thereof.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat vascular dementia. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with vascular dementia.

Seizures, retinal dystrophy, traumatic brain injury, spinal cord injury and long-term depression.

As used herein, retinal dystrophy refers to any disease or condition that is associated with degeneration of the retina. Such diseases or conditions can lead to loss of vision or complete blindness.

As used herein, seizures also include epileptic seizures, and refer to a transient symptom of abnormally excessive or synchronous neural activity in the brain. External manifestations can be dramatic, such as throwing sharp blows, or subtle, such as a momentary loss of perception of reality. Seizures may manifest as changes in mental status, tonic or clonic movements, convulsions, and various other mental symptoms.

Traumatic brain injury (TBI) may also be called intracranial injury. Traumatic brain injuries occur when the brain is traumatically damaged by an external force. Traumatic brain injuries can be classified by severity, mechanism (closed or penetrating head injury), or other characteristics (eg, occurring in a specific location or over a wide area).

Spinal cord injuries (SCI) include any damage to the spinal cord caused by injury rather than disease. Depending on the location of the damage to the spinal cord and nerve roots, symptoms can vary widely, from pain to paralysis and incontinence. Spinal cord injuries are described as having different degrees, and can range from incomplete, with no effect on the patient, to complete injury, meaning complete loss of function.

Long-term depression (LTD) is an activity-dependent decrease in the effectiveness of neuronal synapses, lasting for several hours or longer, after low-frequency (long patterned) stimulation. Long-term depression can occur in many areas of the central nervous system with different mechanisms depending on the brain region and developmental process. Long-term depression can occur in the hippocampus, cerebellum, and in different types of neurons that release different neurotransmitters. Without wishing to be bound by theory, it is believed that long-term depression may be associated with neurodegeneration, dementia and Alzheimer's disease.

Proneurotrophins (e.g. proneurotrophin-4/5, neurotrophin-4/5, pro-NGF, pro-BDNF, etc.) have been shown to play a role in seizures, retinal dystrophy, traumatic brain injury, spinal cord injury and long-term depression.

Accordingly, and without wishing to be limited by theory, it is believed that anti-sortilin antibodies of the present invention that inhibit the interaction between sortilin and the neurotrophins of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro-BDNF , neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.); or that inhibit one or more activities of sortilin, may be used to prevent, reduce the risk of, or treat seizures, retinal degeneration, traumatic brain injury, spinal cord injury, and/or long-term depression in individuals in need thereof.

In some embodiments, administration of an anti-sortilin antibody of the present invention can prevent, reduce the risk of, and/or treat seizures, retinal degeneration, traumatic brain injury, spinal cord injury, and/or long-term depression. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with seizures, retinal dystrophy, traumatic brain injury, spinal cord injury, and/or long-term depression.

Atherosclerotic vascular diseases.

As used herein, atherosclerotic vascular disease, ASVD, and atherosclerosis are used interchangeably and refer to any condition in which the artery wall thickens as a result of the accumulation of fatty materials such as cholesterol, lipids, and triglyceride. Atherosclerotic vascular disease includes, without limitation, any atherosclerotic vascular disease-related condition, disorder or disease, including, without limitation, thromboembolism, stroke, ischemia, infarction, coronary thrombosis, myocardial infarction (eg, heart attack), and intermittent claudication.

- 87 046146

As disclosed herein, the sortilin proteins of the present invention are involved in lipid regulation by binding lipid-associated proteins such as receptor-associated protein, lipoprotein lipase, and the apolipoproteins APOA5 and APOE.

Accordingly, and without wishing to be limited by theory, it is believed that the anti-sortilin antibodies of the present invention that inhibit the interaction between sortilin and lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE), and/or receptor associated protein (RAP); or that inhibit one or more activities of sortilin, can be used to prevent, reduce the risk of, or treat one or more atherosclerotic vascular diseases in individuals in need thereof.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat atherosclerotic vascular disease. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with atherosclerotic vascular disease.

Undesirable symptoms of aging.

As used herein, undesirable symptoms of aging include, but are not limited to, memory loss, behavioral changes, dementia, Alzheimer's disease, retinal degeneration, atherosclerotic vascular disease, hearing loss, and cell destruction.

In some embodiments, and without wishing to be bound by theory, it is believed that anti-sortilin antibodies of the present invention that inhibit the interaction between sortilin and progranulin, the neurotrophins of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro- NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), and/or receptor-associated protein (RAP ); or that inhibit one or more activities of sortilin, may be used to prevent, reduce the risk of, or treat one or more undesirable symptoms of aging.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat one or more undesirable symptoms of aging. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with one or more adverse symptoms of aging.

Amyotrophic lateral sclerosis (ALS).

As used herein, amyotrophic lateral sclerosis (ALS) or motor neurone disease or Lou Gehrig's disease are used interchangeably and refer to a debilitating disease of varying etiologies characterized by rapidly progressive weakness, muscle wasting and fascicular tremor, muscle spasticity, difficulty speaking (dysarthria), difficulty swallowing (dysphagia), and difficulty breathing (shortness of breath).

PGRN (progranulin) haploinsufficiency due to heterozygous loss-of-function mutations in the GRN gene results in decreased PGRN levels in the CSF and is a cause of frontotemporal dementia (FTD) with TDP-43 pathology (Sleegers et al., (2009) Ann Neurol 65:603; Smith et al., (2012) Am J Hum Genet 90:1102). TDP-43 has also been identified as an important pathological protein in ALS, suggesting similarities between ALS and FTD.

For example, more than twenty dominant TDP-43 mutations have been identified in patients with sporadic and familial ALS (Lagier-Tourenne et al., (2009) Cell 136:1001) and TDP-43-positive aggregates are found in approximately 95% of ALS cases (Prasad et al., (2019) Front Mol Neurosci 12:25). In addition, ALS risk genes such as MOBP, C9ORF72, MOBKL2B, NSF, and FUS may also cause FTD (Karch et al., (2018) JAMA Neurol 75:860). In addition, both PGRN and C9ORF72 mutations are associated with abnormal microglial activation, which appears to be another common pathology of FTD and ALS (Haukedal et al., (2019) J Mol Biol 431:1818). Other evidence also suggests that ALS and FTD are closely related conditions with overlapping genetic, neuropathological and clinical features (Weishaupt et al., (2016) Trends Mol Med 22:769; McCauley et al., (2018) Acta Neuropathol 137 :715). Taken together, these results suggest that both diseases may benefit from co-treatment and that genetic variation in PGRN acts as a modifier of ALS progression.

In some embodiments, and without wishing to be bound by theory, it is believed that anti-sortilin antibodies of the present invention that inhibit the interaction between sortilin and progranulin, the neurotrophins of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro- NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), and/or receptor-associated protein (RAP ); or that inhibit one or more activities of sortilin, may be used to prevent or treat one or more adverse symptoms of ALS.

In some embodiments, administration of an anti-sortilin antibody of the present invention

- 88 046146 can prevent, reduce the risk, and/or treat ALS. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with ALS.

Depression.

As used herein, depression or major depressive disorder (MDD), clinical depression, major depression, unipolar depression, unipolar disorder, recurrent depression, or dysthymia are used interchangeably and refer to a mental disorder characterized by episodes of pervasive depressed mood accompanied by low self-esteem and loss of interest in, or pleasure in, normally enjoyable activities.

Accordingly, and without wishing to be limited by theory, it is believed that the anti-sortilin antibodies of the present invention that inhibit the interaction between sortilin and progranulin, the neurotrophins of the present invention (e.g., proneurotrophins, proneurotrophin-3, proneurotrophin-4/5, pro-NGF, pro -BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, lipoprotein lipase (LpL), apolipoprotein AV (APOA5), and/or receptor associated protein (RAP); or that inhibit one or more activities of sortilin, may be used to prevent or treat one or more unwanted symptoms of depression.

In some embodiments, administration of an anti-sortilin antibody of the present invention may prevent, reduce the risk of, and/or treat depression. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with depression.

Parkinson's disease.

Parkinson's disease, which may be called idiopathic or primary parkinsonism, hypokinetic-rigid syndrome (FIRS), or shaking palsy, is a neurodegenerative disorder of the brain that affects the control of the motor system. The progressive death of dopamine-producing brain cells leads to significant symptoms of Parkinson's disease. Most often, Parkinson's disease is diagnosed in people over 50 years of age. Parkinson's disease is idiopathic (no known cause) in most people. However, genetic factors also play a role in the disease.

Symptoms of Parkinson's disease include, but are not limited to, tremors of the hands, arms, legs, jaw and face, stiffness of the muscles of the limbs and trunk, slow movements (bradykinesia), postural instability, difficulty walking, neuropsychiatric problems, changes in speech or behavior, depression, anxiety, pain, psychosis, dementia, hallucinations and sleep problems.

In some embodiments, administration of an anti-sortilin antibody of the present invention can prevent, reduce the risk of, and/or treat Parkinson's disease. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with Parkinson's disease. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with Parkinson's disease.

Huntington's disease.

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by an autosomal dominant mutation in the huntingtin gene (HTT). Expansion of the cytokine-adenine-guanine (CAG) triplet repeat in the huntingtin gene results in the production of a mutant form of the huntingtin (Htt) protein encoded by the gene. This mutant huntingtin protein (mHtt) is toxic and is involved in neuronal death. Symptoms of Huntington's disease most often appear between ages 35 and 44, although they can occur at any age.

Symptoms of Huntington's disease include, but are not limited to, motor control problems, jerky, chaotic movements (chorea), abnormal eye movements, balance problems, seizures, difficulty chewing, difficulty swallowing, cognitive problems, altered speech, memory deficits, difficulty speaking. thinking, insomnia, fatigue, dementia, personality changes, depression, anxiety, and compulsive behavior.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat Huntington's disease. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with Huntington's disease. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with Huntington's disease.

Tauopathy.

Taupathy diseases, or tauopathies, are a class of neurodegenerative diseases caused by the aggregation of microtubule-associated protein tau in the brain. Bo

- 89 046146 Alzheimer's disease (AD) is the best known disease of the tauopathies and involves the accumulation of tau protein in neurons in the form of insoluble neurofibrillary tangles (NFTs). Other tauopathies and disorders include progressive supranuclear palsy, boxer's dementia (chronic traumatic encephalopathy), frontotemporal dementia and parkinsonism associated with chromosome 17, Lytico-Bodig disease (parkinsonism-dementia complex in Guam), dementia with predominant tangles, ganglioglioma and gangliocytoma, meningoangiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease, lipofuscinosis, Pick's disease, corticobasal degeneration, argyrophilic granule disease (AGD), Huntington's disease, and frontotemporal lobar degeneration.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat tauopathy. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with tauopathy. In some embodiments, administration of an anti-sortilin antibody may modulate one or more sortilin activities in an individual with tauopathy.

Multiple sclerosis.

Multiple sclerosis (MS) may also be called disseminated sclerosis or disseminated encephalomyelitis. MS is an inflammatory disease that damages the fatty myelin sheaths around axons in the brain and spinal cord, leading to demyelination and scarring and a wide range of signs and symptoms.

Symptoms of PC include, but are not limited to, changes in sensation such as loss of sensation or tingling; stabbing pain or numbness such as hypoesthesia and paresthesia; muscle weakness; clonus; muscle spasms; difficulty moving; complications with coordination and balance such as ataxia; problems with speech, such as dysarthria, or swallowing, such as dysphagia; vision problems such as nystagmus, retrobulbar neuritis including phosphenes, and diplopia; fatigue; acute or chronic pain; and problems with the bladder and intestines; cognitive impairment of varying degrees; emotional symptoms of depression or unstable mood; Uthoff's phenomenon, which is an exacerbation of observed symptoms when exposed to temperatures above normal ambient temperature; and Lhermitte's sign, which is an electrical sensation running down the back when the neck is flexed.

In some embodiments, administration of an anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat multiple sclerosis. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with multiple sclerosis. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with multiple sclerosis.

Glaucoma and macular degeneration.

Glaucoma describes, without limitation, a group of diseases characterized by damage to the optic nerve, leading to vision loss and blindness. Glaucoma is usually caused by increased fluid pressure (= intraocular pressure) in the anterior chamber of the eye under the cornea. Laucoma results in progressive loss of retinal ganglion cells, which are important for vision. Age-related macular degeneration usually affects older people and primarily causes vision loss in the macula, the central field of vision. Macular degeneration causes, but is not limited to, drusen, pigmentation changes, vision distortions, ocular bleeding, atrophy, decreased visual acuity, blurred vision, central scotomas, decreased color vision, and decreased contrast sensitivity.

Without wishing to be bound by theory, it is believed that administration of the anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat glaucoma and macular degeneration. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with glaucoma or macular degeneration. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with glaucoma or macular degeneration.

Degenerative disc disease (DDD).

Degenerative intervertebral disc disease (DDD) describes, without limitation, a group of diseases in which the intervertebral disc (IVD) undergoes significant morphological as well as biomechanical changes, and usually manifests clinically in patients with low back pain. Degenerative discs typically demonstrate degenerative fibrocartilaginous tissue and chondrocyte clusters suggesting repair. Inflammation may or may not be present. Pathological findings in DDD include protrusion, spondylolysis, and/or vertebral subluxation (sponylolisthesis) and spinal stenosis.

Without wishing to be bound by theory, it is believed that administration of the anti-sortilin antibody of this invention can prevent, reduce the risk of, and/or treat degenerative diseases

- 90 046146 lefting of intervertebral discs. In some embodiments, administration of an anti-sortilin antibody may induce one or more progranulin activities in an individual with degenerative intervertebral disc disease. In some embodiments, administration of an anti-sortilin antibody may modulate one or more activities of sortilin in an individual with degenerative intervertebral disc disease.

Pain.

Pain describes, without limitation, neuropathic pain resulting from nerve damage, such as injury or disease. Such injuries may include damage to peripheral nerves and/or the spinal cord. Sortilin antagonists have been shown to relieve neuropathic pain. (Richner M et al. (2019) Sci. Adv. 5: eaav9946.)

Sets/products.

Provided herein are products (eg, kits) containing the anti-sortilin antibody described herein. The article may include one or more containers containing an antibody described herein. Containers can be any suitable packaging, including, but not limited to, vials, bottles, jars, flexible packaging (eg, sealed Mylar or plastic bags), and the like. Containers may be unit doses, bulk packages (eg, multi-dose packages), or sub-unit doses.

In some implementations, the kits may further comprise a second agent. In some embodiments, the second agent is a pharmaceutically acceptable buffer or diluent, including, without limitation, bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution, and dextrose solution. In some embodiments, the second agent is a pharmaceutically active agent.

In some embodiments of any articles, the articles further include instructions for use in accordance with the methods of the present invention. The instructions typically include information regarding the dosage, dosage regimen, and route of administration of the intended treatment. In some embodiments, such instructions include a description of administering an isolated antibody of the present invention (e.g., an anti-sortilin antibody described herein) to prevent, reduce the risk of, or treat an individual with a disease, disorder, or lesion selected from dementia, frontotemporal dementia , Alzheimer's disease, Gaucher's disease, vascular dementia, epileptic seizures, retinal dystrophy, traumatic brain injury, spinal cord injury, atherosclerotic vascular disease, unwanted symptoms of normal aging, amyotrophic lateral sclerosis (ALS), long-term depression, Parkinson's disease, Huntington's disease, tauopathy, multiple sclerosis, age-related macular degeneration, glaucoma, degenerative disc disease (DDD), Creutzfeldt-Jakob disease, normal pressure hydrocephalus, Nasu-Hakola disease, stroke, acute trauma, chronic trauma, lupus, acute and chronic colitis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, malaria, essential tremor, lupus of the central nervous system, Behçet's disease, mixed dementia, dementia with Lewy bodies, multiple system atrophy, Shy-Drager syndrome, progressive supranuclear palsy, corticobasal ganglion degeneration, acute disseminated encephalomyelitis, granulomatous disorders, sarcoidosis, diseases of aging, retinitis pigmentosa, retinal degeneration, respiratory tract infection, sepsis, eye infection, systemic infection, lupus, arthritis and wound healing, in accordance with any methods of this invention. In some embodiments, the disease, disorder, or lesion is frontotemporal dementia. In some embodiments, the instructions include instructions for using an anti-sortilin antibody and a second agent (eg, a second pharmaceutically active agent).

Diagnostic applications.

Isolated antibodies of this invention (eg, the anti-sortilin antibody described herein) are also suitable for diagnostic use. This disclosure therefore provides methods for using an antibody of this invention, or functional fragments thereof, for diagnostic purposes, such as detecting the protein sortilin in an individual, or in tissue samples obtained from an individual.

In some embodiments, the individual is a human. In some embodiments, the individual is a patient suffering from, or at risk of developing, a disease, disorder or impairment of the invention. In some embodiments, the diagnostic methods involve detecting sortilin protein in a biological sample, such as a biopsy sample, tissue, or cells. The anti-sortilin antibody described herein is contacted with a biological sample and antigen-bound antibody is detected. For example, a biopsy sample may be stained with the anti-sortilin antibody described herein in order to detect and/or quantify disease-associated cells. The detection method may include quantifying antigen-associated antigen

- 91 046146 bodies. Detection of antibodies in biological samples can be performed by any method known in the art, including immunofluorescence microscopy, immunocytochemistry, immunohistochemistry, ELISA, FACS (fluorescence activated cell sorting) analysis, immunoprecipitation, or micro-positron emission tomography. In some embodiments, the antibody is labeled with a radioactive isotope, such as ¹⁸ F, and then detected using micropositron emission tomography analysis. Antibody binding can also be quantified in a patient by non-invasive techniques such as positron emission tomography (PET), X-ray computed tomography, single photon emission computed tomography (SPECT), computed tomography (CT), and axial computed tomography (ACT).

In other embodiments, an isolated antibody of the present invention (e.g., the anti-sortilin antibody described herein) can be used to detect and/or quantify, for example, microglia in a brain sample taken from a preclinical disease model (e.g., not -human model of the disease). As such, an isolated antibody of the present invention (e.g., the anti-sortilin antibody described herein) may be useful for assessing therapeutic response after treatment in a model of a disease or lesion of the nervous system, such as frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, atherosclerotic vascular disease, Nasu-Hakola disease, or multiple sclerosis, compared with controls.

The present invention will be better understood with reference to the following Examples. However, they should not be construed as limiting the scope of the present invention. All documents referred to herein are hereby expressly incorporated herein by reference.

Examples

Examples 1-3 describe the preparation of affinity matured variants of the anti-human sortilin antibody, S-60, and the characterization of these variants not only for their SORT1 binding affinity, but also for their biological activity in reducing SORT1 expression, increasing PGRN secretion, and blocking binding PGRN-SORT1. Surprisingly, it has been found that increased biological activity does not necessarily correlate with increased affinity.

Example 1: Generation of S-60 variants and measurement of SORT1 binding affinity.

The purpose of the following Example was to generate affinity matured variants of the anti-human sortilin antibody, S-60, and to characterize the binding of the affinity matured antibody to human sortilin (SORT1).

Affinity matured S-60 variants.

Affinity-matured anti-human sortilin (SORT1) antibodies were generated and their binding to SORT1 was measured. Antibody against SORT1, S-60, as well as S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60- 7, S-60-8, and S-60-9 were described in WO2016164637. The VH and VL sequences of S60-9 and S60 in WO2016164637 are identical, and S-60 will be used herein in relation to the antibody. The VH and VL sequences of S-60-5, S-60-6 and S-60-7 in WO2016164637 are identical, and S-60-7 will be used herein in relation to the antibody. S-60 was found to be both a strong regulator of SORT1 levels and a strong blocker of binding between SORT1 and PGRN (see, for example, Tables 8-10 and Figures 6A and 10D in WO2016164637). Therefore, S-60 was chosen for further affinity maturation.

Anti-SORT1 variants S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15, S-6016, S-60-18, S-60-19, and S-60-24 were obtained as described below.

Production of affinity matured S-60 variants.

Anti-SORT1 antibody S-60 was affinity matured. Briefly, diversified antibody libraries were generated in yeast for each primary parent antibody. Diversity in the first round of affinity maturation was generated by using standard molecular cloning techniques to combine the original HVR-H3 heavy chain and light chain (LC) with specified genetic diversity in the HVR-H1 and HVR-H2 regions of the heavy chain (HC) (termed H1/H1 optimization). H2). For the second round of affinity maturation, both the variable heavy chain (VH) and variable light chain (VL) sequences were optimized, with a particular focus on HVR-H3.

The selection pressures used to screen the library included equilibrium titration of the human SORT1 antigen, competitive Fab kinetics of the parent antibody, and the use of a polyspecificity deselection reagent. FACS flow cytometry was then used to visualize and select antibodies using standard techniques (see, for example, Chao et al. Nature Protocols, 2006). The desired population was then selected for additional rounds of selection.

Anti-SORT! Affinity-matured antibodies were purified as follows: clones were grown to saturation and then induced for 48 h at 30°C with shaking. After induction, yeast cells were pelleted by centrifugation and supernatants were collected for purification. Immunoglobulins were purified using a protein A column and eluted with acetic acid, pH 2.0. Fab

- 92 046146 fragments were prepared by papain digestion and purified using a CaptureSelect IgG-CH1 affinity matrix (LifeTechnologies).

Measurement of binding affinity to SORT 1.

Anti-SORT1 antibody S-60 and its affinity matured variants were tested and compared using a SORT1 binding assay. For binding assays, stable HEK293T cells expressing human SORT1 were harvested by trypsinization, washed with PBS, counted, and seeded into 96-well U-bottom plates at 1 x 105 cells/well. The plates were centrifuged at 1400 rpm for 3 minutes and primary anti-SORTI or control antibodies in FACS buffer (PBS+2 FBS) were added and incubated on ice for one hour. Cells were then centrifuged as previously described and washed three times with FACS buffer. Cells were then incubated with anti-human APC-conjugated secondary antibody (BD Biosciences) in FACS buffer for 30 min on ice. Cells were washed again three times with FACS buffer and analyzed using a BD FACS Canto or an Intellicyt flow cytometer. Binding was measured as the median fluorescent intensity (MFI) in the PE channel of a population of GFP-positive cells.

Results.

The binding affinities of the anti-SORT1 antibody to SORT1 are shown in Table 1 below. 1. All antibodies were tested on the huIgG1 scaffold.

*Note: KD values represent the average of two experiments.

Anti-SORTI antibodies S-60-3, S-60-4, S-60-7, S-60-8, S-60-10, S-60-11, S-60-13, S-60- 15, S-60-16, S60-19, and S-60-24 showed significantly increased binding affinity for SORT1 compared to S-60. Specifically, S-60-8 and S-60-15 exhibited the highest binding affinities for SORT1 of the variants tested, with 2.46- and 2.05-fold increased values, respectively, compared to the binding affinities of the parent antibody, S- 60.

Example 2. Effect of S-60 variants on SORT1 expression and PGRN secretion.

The purpose of the following Example was to characterize the effect of affinity matured variants of the human anti-SORT1 antibody S-60 on SORT1 expression and extracellular progranulin (PGRN) levels.

Analyzes of SORT1 expression and PGRN secretion.

Affinity mature S-60 variants were generated as described in Example 1 and then screened by FACS for their ability to reduce SORT1 expression and increase extracellular PGRN levels. To evaluate the effect of anti-SORT1 antibodies on SORT1 expression and PGRN secretion, U251 cells, which endogenously express SORT1 and secrete PGRN, were incubated with anti-SORT1 antibody S-60 and affinity-matured variants. Cell surface levels of SORT1 at different antibody concentrations were measured by FACS as follows: S-60 antibody against

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SORT1 and affinity matured variants were added to U251 cells seeded at 3x10 ³ cells/well in 96-well plates, with serial dilutions ranging from 0.023 nM to 150 nM. After 72 h, cells were harvested with trypsin, washed with PBS, and labeled with Dylight-650-conjugated anti-SORT1 antibody S-2-11 (described in WO2016164637) to quantify residual SORT1 protein levels. After incubating cells with 5 μg/ml S-2-11 conjugated to DyLight-650 (Invitrogen) for one hour on ice, cells were washed three times in PBS+ 2 FBS and binding was quantified using a FACSCanto™ or an Intellicyt flow cytometer as median fluorescence intensity (MFI) of APC.

The amount of PGRN secreted into the cell supernatant over 72 h was measured by standard ELISA as follows: harvest the cells and collect the medium; PGRN concentrations in media samples were measured using the Duoset human PGRN ELISA kit from R&D Systems, according to the manufacturer's instructions. Data were analyzed using Microsoft Excel and GraphPad Prism.

Results.

The half-maximal effective concentration (EC ₅₀ ) of down-regulation (DR) of cell surface SORT1 levels for each of the S-60 variants tested is shown in Table. 2. Reduction in SORT1 expression (DR) was quantified for each S-60 variant tested and expressed as a percentage of control (untreated cells) (Tables 2-4). Additionally, the level of extracellular secretion of PGRN was quantified and expressed as fold change compared to the untreated control, as indicated in Table. 2-4. The results of independent experiments are given in separate tables. (Table 2-4). All antibodies were tested on the huIgG1 WT (wild type) scaffold.

table 2

Percentage reduction (DR) of SORT1 expression and PGRN secretion for S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-7 and S-60-8

Antibody designation EC50 expression reduction (M) Reduced expression at 0.6 nM antibody (IgG) PGRN secretion (fold compared to control) 0.62 or 0.63 nM antibody (IgG) S-60 6.81E-10 29.3 1.38 S-60-1 2.73E-10 50.4 1.60 S-60-2 4.20E-10 40.3 1.75 S-60-3 2.69E-10 43.0 1.77 S-60-4 4.18E-10 39.6 1.61 S-60-7 4.47E-10 32.6 1.40 S-60-8 5.76E-10 32.0 1.54

Table 3

Percentage reduction (DR) of SORT1 expression and PGRN secretion for S-60, S-60-3, S-60-24, S-60-15, S-60-16, S-60-18 and S-60-19

Antibody designation Reduced expression at 1.25 nM antibody (IgG) PGRN secretion (fold compared to control), 0.63 nM antibody (IgG) S-60 40.17 1.11 S-60-3 61.50 1.32 S-60-24 65.20 1.35 S-60-15 60.92 1.42 S-60-16 70.14 1.42 S-60-18 51.33 1.13 S-60-19 55.46 1.35

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Table 4

Percentage reduction (DR) of SORT1 expression and PGRN secretion for S-60-24, S-60-10, S-60-11, S-60-12, S-60-13 and S-60-14

Antibody designation Reduced expression at 0.63 nM antibody (IgG) PGRN secretion (fold compared to control), 0.63 nM antibody (IgG) S-60-24 81.48 1.62 S-60-10 85.41 1.85 S-60-11 88.76 1.89 S-60-12 87.38 1.89 S-60-13 82.33 1.77 S-60-14 80.47 1.81

In the first experiment analyzing the EC50 reduction in SORT1 expression, percentage reduction in SORT1 expression using a saturating antibody concentration of 0.6 nM IgG, and PGRN secretion using a saturating antibody concentration of 0.62 or 0.63 nM IgG, the following were tested: options: S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, and S-60-8. As indicated in table. 2, S-60-3 showed the smallest EC50 decrease in expression (0.269 nM) and the highest PGRN secretion, 1.77 times greater than the control, and better than S-60, which showed a 1.38 times increase in compared to the control. When using 0.6 nM IgG, S-60-1 showed the most significant effect on SORT1 expression of the tested variants, reducing SORT1 expression by 50.4% compared to control, while S-60 reduced SORT1 expression by 29.3 %.

In the following experiment, the percentage reduction in expression using a saturating antibody concentration of 1.25 nM IgG and PGRN secretion using a saturating antibody concentration of 0.63 nM IgG were analyzed for the following variants: S-60, S-60-3 , S-6024, S-60-15, S-60-16, S-60-18, and S-60-19. As indicated in table. 3, both S-60-15 and S-60-16 showed the highest levels of extracellular PGRN secretion with a 1.42-fold increase compared to control, which was higher than S-60, which showed a 1.11-fold increase in compared to the control. Indeed, these two variants also significantly reduced SORT1 expression compared to S-60, reducing SORT1 expression by 60.92% and 70.14%, respectively, compared to control, while S-60 reduced SORT1 expression by only 40.17%.

In the third experiment, the percentage reduction in expression using a saturating antibody concentration of 0.63 nM IgG and PGRN secretion using a saturating antibody concentration of 0.63 nM IgG were analyzed for the following variants: S-60-24, S-60 -10, S-60-11, S60-12, S-60-13 and S-60-14. As indicated in table. 4, S-60-11 and S-60-12 showed the highest amount of extracellular PGRN secretion with a 1.89-fold increase compared to the control. S-60-11 also showed the highest reduction in SORT1 expression at 0.63 nM IgG, reducing SORT1 expression by 88.76% compared to control.

Conclusions.

As indicated in table. 1 in Example 1, S-60-8 and S-60-15 showed the highest binding affinities for SORT1 compared to S-60 and other S-60 variants tested, with the lowest KD values of 4.68E-1OM and 5. 60E-10, respectively. However, when compared with a specific variant, S-60-3, which has a higher KD value of 8.56E-1OM (Table 1), S-60-8 showed a smaller increase in PGRN secretion (Table 2), while while S-60-15 showed a greater fold increase in PGRN secretion than S-60-3 (Table 3). Additionally, S-60-8 (at a saturating concentration of 0.6 nM IgG) showed a smaller percentage reduction in SORT1 expression (Table 2) compared to S-60-3 and S-60-15 (at a saturating concentration of 1.25 nM IgG), which reduced SORT1 expression by 61.50% and 60.92%, respectively (Table 3). In summary, S-60-15 showed better results than S-60-3 in increasing PGRN secretion, and showed a similar effect as S-60-3 in reducing cell surface SORT1 levels, while the results for S- 60-8 were worse than S-60-3 in terms of increased PGRN secretion and decreased cell surface SORT1 levels.

The results of this study indicate that the relative potency of anti-SORT1 antibody S-60 variants in terms of decreased SORT1 expression or increased PGRN secretion cannot be predicted by their relative binding affinities for sortilin.

In addition, S-60-15 showed the greatest increase in PGRN secretion compared with S60 and certain S-60 variants (Table 3, last column), including S-60-3. S-60-3, in turn, showed increased secretion of PGRN compared to S-60-1, -2, -4, -7, and -8 (Table 2, last column). Thus, S-60-15 is an effective antibody for increasing

- 95 046146 PGRN secretion compared to these S-60 variants. The ability to increase PGRN secretion is an important property when selecting a therapeutic antibody for the treatment of diseases such as FTD and other pathologies in which PGRN deficiency plays a direct causative role. Such an antibody would be useful in restoring PGRN levels to, or close to, wild-type levels, thereby curing the disease. In addition, this property is an important pharmacodynamic parameter when assessing the activity of an antibody in vivo (see below, example 5).

Example 3: S-60 variants block PGRN binding to SORT1.

The purpose of the following Example was to characterize the effect of affinity matured variants of the anti-human SORT1 antibody S-60 on the binding interaction between SORT1 and the natural ligand PGRN.

SORT1-PGRN blocking assay.

The ability of S-60 variants to block PGRN binding to SORT1 was also tested. Recombinant human PGRN (PGRN) (Adipogen) was biotinylated using the EZ-Link Micro NHS-PEG4 kit from ThermoScientific/Pierce according to the manufacturer's instructions. A stable cell line expressing full-length untagged human SORT1 (SORT1) was generated by viral infection of HEK293T cells and positive selection with hygromycin (a special Genscript project). Parental HEK293T cells were used as control cells.

SORT1-expressing cells or control cells were collected and washed with PBS. Biotinylated human PGRN was added in PBS+2% FBS with or without titrated anti-SORT1 antibodies or human IgG1 isotype control antibodies and incubated on ice for 2 hours. After washing the cells 3 times in PBS+2% FBS, the cells were incubated in streptavidin -APC (BD Biosciences, 1:100) on ice for 30 min. Cells were then washed again, resuspended in PBS+2% FBS, and analyzed on a FACSCanto™ flow cytometer (BD Biosciences, Mississauga, ON). PGRN binding was measured as the median fluorescence intensity (MFI) of APCs in a population of SORT1-expressing cells. The binding of biotinylated PGRN to SORT1 expressed on HEK293T cells was measured by FACS in the absence or presence of antibody S-60 and its variants.

Results.

The half-maximal effective concentration (EC50) for blocking the binding of PGRN to SORT1 for each of the tested S-60 variants is given in table. 5-7. Additionally, the maximum level of blocking of PGRN by SORT1 was quantified and expressed as a percentage of the maximum blocking achieved using a saturating concentration of 50 nM IgG or 150 nM IgG (Tables 5-7). Independent experiments are shown in separate tables below.

Table 5

Blocking the binding of progranulin (PGRN) to SORT1 expressed on HEK293T cells, ____________for S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, S-60-8 and S-60____________

Antibody designation EC ₅₀ blocking PGRN binding (nM) Blocking PGRN binding (% of maximum at 150 nM antibody (IgG)) S-60-1 5,986 95.2 S-60-2 7,509 94.5 S-60-3 0.733 95.5 S-60-4 0.584 95.7 S-60-7 0.944 96.5 S-60-8 0.985 95.6 S-60 0.751 95.9

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Table 6 Blocking the binding of progranulin (PGRN) to SORT1 expressed on HEK293T cells, _________for S-60, S-60-3, S-60-24, S-60-15, S-60-16, S-60-18 and S-60-19_________

Antibody designation EC ₅₀ blocking PGRN binding (M) Blocking PGRN binding (% of maximum at 50 nM antibody (IgG)) S-60 2.20E-09 90.13 S-60-3 1.76E-09 90.44 S-60-24 1.11E-09 88.94 S-60-15 1.22E-09 90.74 S-60-16 1.20E-09 90.44 S-60-18 2.27E-09 89.33 S-60-19 1.53E-09 90.15

Table 7 Blocking the binding of progranulin (PGRN) to SORT1 expressed on HEK293T cells, ___________for S-60-24, S-60-10, S-60-11, S-60-12, S-60-13 and S-60 -14___________

Antibody designation EC ₅₀ blocking PGRN binding (M) Blocking PGRN binding (% of maximum at 50 nM antibody (IgG)) S-60-24 4.70E-10 95.64 S-60-10 4.60E-10 93.84 S-60-11 4.10E-10 94.97 S-60-12 6.10E-10 93.26 S-60-13 5.60E-10 93.83 S-60-14 6.00E-10 93.67

In the first experiment analyzing the EC ₅₀ of blocking PGRN binding and the percentage of blocking progranulin (PGRN) binding to SORT1 at a saturating concentration of 150 nM IgG, the following variants were tested: S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, S-60-8, and S-60. As indicated in table. 5, S-60-4 exhibited the lowest EC ₅₀ value (0.584 nM), achieving 95.7% blockade of progranulin (PGRN) binding to SORT1. Of the variants analyzed, S-60-7 showed the greatest reduction in PGRN binding, blocking 96.5% of progranulin (PGRN) binding to SORT1, although it showed lower efficiency in blocking PGRN binding (higher EC50 value) than S-60 and variants S-60-3 and S-60-4.

In the following experiment, analyzing the binding of PGRN EC ₅₀ and the percentage of blocking of progranulin (PGRN) binding to SORT1 at a saturating concentration of 50 nM IgG, the following variants were tested: S-60, S-60-3, S-60-24, S- 60-15, S-60-16, S-60-18, and S-60-19. As indicated in table. 6, S-60-24, S-60-15 and S-60-16 showed the lowest EC ₅₀ compared to S-60, with values ranging from 1.11 nM to 1.22 nM. In addition, the S-60-15 variant showed the greatest activity compared to S-60, achieving 90.74% blockade of progranulin (PGRN) binding to SORT1.

In the third experiment, the following options were tested: S-60-24, S-60-10, S-60-11, S60-12, S-60-13 and S-60-14. As indicated in table. 7, S-60-11 showed the lowest EC ₅₀ (0.410 nM), achieving 94.97% blocking progranulin (PGRN) binding to SORT1. Of the variants analyzed, S-60-24 showed the greatest reduction in PGRN binding, blocking 95.64% of progranulin (PGRN) binding to SORT1.

Conclusions.

As indicated in table. 1 in Example 1, both S-60-8 and S-60-15 demonstrated the highest binding affinities for SORT1 compared to S-60 and other S-60 variants tested, with S-60-8 having the highest affinity for SORT1 . However, when compared with S-60, S-60-8 showed lower efficiency in blocking PGRN binding (higher EC ₅₀ value compared to S-60) (Table 5), while S-60-15 demonstrated higher efficiency in blocking binding to PGRN (lower EC50 compared to S-60) (Table 6). In addition, S

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60-8 showed a lower percentage of maximal blocking of progranulin (PGRN) binding to SORT1 compared to S-60 (Table 5), while S-60-15 showed a higher percentage of maximal blocking of progranulin (PGRN) binding to SORT1 to compared to S60 (Table 6). Thus, S-60-15 performed better than S-60-8 compared to S-60 in terms of EC ₅₀ value in blocking PGRN binding to SORT1 and maximum percentage blocking PGRN binding to SORT1, despite the fact that S-60-8 showed higher binding affinity for SORT1 compared to S-60-15.

Thus, the results of these studies unexpectedly showed that the relative potency of anti-SORT1 antibody S-60 variants in blocking the extent of progranulin (PGRN) binding to SORT1 cannot be predicted by their relative binding affinity to SORT1.

Example 4: Stability of S-60-15 and stress test analysis.

The purpose of the following Example was to determine the stability of S-60-15, which is the affinity matured version of S-60 described in the previous Examples, under different stress test conditions.

The stability of a therapeutic antibody is important for clinical effectiveness. During production, storage and in vivo administration, therapeutic antibodies are at risk of degradation in various ways. The factors responsible for such degradation remain poorly understood. Given that S-60-15 demonstrated high binding affinity to SORT1 (see Example 1), effectively reduced cell surface SORT1 levels and increased extracellular secretion of PGRN (see Example 2), and also significantly blocked PGRN binding to SORT1 (see Example 1). Example 3), and unexpectedly passed these functional tests with better results than S-60-8, which showed higher affinity for SORT1, S-60-15 was further analyzed for stability.

pH and temperature stress testing.

To measure the stability of amino acid residues in S-60-15, samples of S-60-15 were exposed to stress conditions of pH and temperature to simulate the stress conditions that may occur during production, storage, and in vivo administration. Briefly, individual S-60-15 samples were stress tested at pH 3.5 or pH 5.0 and at 40°C or 50°C. Samples were tested at different time points - 0, 1, 3 and 5 days for two pH conditions, and 0, 7, 14 and 30 days for high temperature stress conditions - by treatment with DTT (dithiothreitol) and IAA (indolylacetic acid) followed by trypsin digestion . The hydrolyzed samples were analyzed by liquid chromatography with mass spectroscopic detection using a Waters ACQUITY UPLC coupled to a Xevo G2-XS QTOF mass spectrometer using a BEH C18 column. The results are shown in table. 8.

Table 8

Stoichiometric percentage table (% of each peptide with modification). Deamid (deamidation); Oxid (oxidation); CAM (carbamidomethyl)

S-60-15 Peptide Modification Without stress pH5 (10 pH5 dl pH5 (13 pH5 d5 pH3.5 dO pH3 _: 5 dl pH3.5 (13 pH3.5 d5 T4G d7 T40 dl4 T40 (130 T50 d7 T50 dl4 T50 d30 NQFSLK (SEQ ID NO:87) Deamid 1.3 1.5 1.5 1.5 0.9 1.5 1.4 1.5 1.4 1.3 1.3 1.3 1.3 1.3 1.4 QGYYGMDVWGQGTTVTVSSASTK (SEQIDNO:88) Oxide M(l) 62 6.8 6.2 6.6 8.8 6.2 6.8 6.6 6.4 5.7 5.1 5.6 5.1 5.4 1.3 QGYYGMDVWGQGTTVT VSSASTK (SEQ ID NO:88) Deamid Q(i) θ,5 0.6 0.6 0.6 0.7 0.6 0.6 0.6 0.6 0.8 1.0 1.0 1.4 2.3 3.6 DYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSWTV PSSSLGIK (SEQ ID NO:89) Deamid N(l) Oxide W(l) 3.3 3.3 3.3 3.3 3.8 3.4 3.4 3.4 3.2 3.2 3.1 0.0 3.2 3.1 3.2 DYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSWTV PSSSLGIK (SEQ ID NO:89) Deamid N(l) 0.3 0.4 0.4 0.4 0.4 0.4 0.4 0.3 0.3 0.4 0.5 0.8 0.7 1.0 1.5 DTLMISR (SEQ ID NO:90) Oxide M(l) 3.1 3.6 3.4 3.5 3.5 3.6 3.1 3.4 3.3 4.0 4.4 5.8 5.2 7.2 12.2 TPEVTCVWDVSQEDPEV QFNWYVDGVEVHNAK (SEQIDNO:91) Deamid N(l) 1.4 1.1 1.2 1.3 1.7 1.5 1.2 1.4 1.3 0.2 0.3 1.3 1.1 0.2 5.2 WSVLTVLHQDWLNGK (SEQIDNO:92) Deamid N(l) 7.8 7.2 6.7 6.9 6.5 7.0 6.9 7.0 6.8 8.1 8.4 9.2 9.3 12.6 12.0 EPQVYTLPPSQEEMTK (SEQIDNO:93) Oxid. M(l) 0.3 0.3 0.0 0.0 1.5 0.2 0.4 0.4 0.5 0.8 0.0 0.0 0.8 1.3 1.1

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NQVSLTCLVK (SEQ ID NO:94) Deamid N(l) CAM €(1) 1.9 2.1 2.0 2.0 1.7 2.0 2.0 2.0 1.9 1.9 2.1 1.9 2.1 2.5 2.0 GFYPSDIAVEWESNGQPENNYK (SEQIDNO:95) Deamid N(l) V 4.9 5.1 5.0 6.4 5.0 5.0 5.2 4.9 7.0 8.9 9.6 H,1 19.0 45.9 WQEGNVFSCSVMHEALH NHYTQK (SEQ ID NO:96) Deamid N(l) CAM €(1) 3.8 4.0 4.0 4.0 4.4 3.9 3.0 4.0 3.9 3.7 3.7 3.9 3.9 4.4 6.7 WQEGNVFSCSVMHEALH NHYTQK (SEQ ID NO:96) CAM €(1), Oxid. M(l) 1.6 1.8 1.8 1.8 1.9 1.8 1.8 1.9 1.8 1.8 1.9 2.6 2.2 2.6 5.1 DIVMTQSPLSLPVTPGEPASISCR (SEQIDNO:97) CAM €(1), Oxid. M(l) 0.7 0.8 0.8 0.8 0.8 0.8 0.7 0.8 0.7 0.8 0.8 1.1 0.8 0.8 2.0 SSQSLLR (SEQ ID NO:98) Deamid QCD 6.8 6.1 5.2 5.4 6.5 6.0 5.4 6.7 5.8 7.3 6.9 7.0 6.8 7.2 6.6 SNGYNYLDWYLQKPGQSP QLLIYLGSNR (SEQ ID NO:99) Deamid N(l) 11.2 9.3 9.4 9.3 12.4 9.8 9.7 9.8 9.6 15.7 2O _? 3 21.9 30.7 45.4 65.6 AEAEDVGVYYCMQQQEAP LTFGGGTK (SEQ ID NO: 100) CAM C(l), Oxide. M(l) 0.5 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.5 0.6 1.1 0.8 1.1 2.1 SGTASWCLNNEYPR (SEQ ID NOT 01) Deamid N(l) 2.6 2.8 2.9 2.8 3.1 2.8 2.7 2.8 2.7 2.6 2.7 2.5 2.6 3.0 3.4 VDNALQSGNSQESVTEQDSK (SEQIDNO:102) Deamid N(l) 0.3 0.4 0.4 0.4 0.4 0.3 0.4 0.4 0.4 0.5 0.5 0.5 0.6 0.8 1.3

Amino acids potentially subject to post-translational modifications (PTMs) under stress were identified and corresponding peptide fragments obtained from stressed samples were monitored kinetically. The results confirmed that two asparagine-glycine sites, one in the heavy chain Fc region (SEQ ID NO: 11) and one in the light chain Fab region (SEQ ID NO: 15) of S60-15, were most susceptible to PTM, demonstrating 45.9% and 65.6% deamidation, respectively, after storage at 50°C for 30 days. No other post-translational modification or cleavage site showed significant post-translational modification or degradation in stress tests.

Deamidation of the asparagine site (N33).

Any deamidation in the Fc region of the S-60-15 heavy chain should not affect the binding affinity of S-60-15 to SORT1, since this region does not interact directly with SORT1. However, the asparagine site (N33) in the Fab region of S-60-15 is located in HVR-L1 (SEQ ID NO: 8), a light chain complementarity determining region that is involved in antigen binding. Thus, deamidation of the asparagine site (N33) in the Fab region of S-60-15 may influence the binding affinity of S-60-15 to SORT1. Upon deamidation, the asparagine site (N33) in the Fab region of S-60-15 undergoes an Asn substitution with Asp/u30-Asp.

Site-directed mutagenesis was used to test the effects of deamidation of the asparagine site (N33) in the S-60-15 Fab region and potential amino acid substitution to reduce the likelihood of degradation and deamidation and the resulting production consequences. Seventeen different point mutations at position N33 of HVR-L1 S-60-15 huIgG1 were generated and sequenced using standard procedures. The Glm3 or Glm(f) allotype with L234A/L235A/P331S mutations (LALAPS motif) was used as background (Jefferis R and Lefranc M-P, Mabs, 2009 Jul-Aug; 1(4): 332-338). LALAPS point mutations are intended to improve overall safety by minimizing effector functions such as Fc-gamma receptor binding, complement activation, and antibody-mediated cellular cytotoxicity (ADCC). Reduced Fc gamma receptor binding, complement activation, and ADCC as a result of LALAPS mutations have been observed in various experiments (data not shown). Biolayer interferometry (BLI) data were collected at 5 Hz at 25°C on a Pall ForteBio Octet RED96 instrument. Data analysis was performed using the data analysis application ForteBio Data Analysis Software, version 9.0. Standard kinetic buffer (PBS, 0.1 BSA, 0.02 Tween-20, pH 7.2) was used for analysis and for reagent preparation. For all assays, probe tips were equilibrated in buffer prior to analysis.

For the primary screen for dissociation rate, mutants S-60-15 LALAPS N33 and N33X (1

- 99 046146 μg/ml, loading time 300 s) was captured on Anti-Human IgG Fc Capture Dip and Read Biosensors (Pall ForteBio, Menlo Park, CA). 20 nM histidine-tagged human SORT1 (R&D Systems, Minneapolis, MN) was then bound to surface-captured anti-SORT1 (binding time 200 s, dissociation time 1200 s). The resulting BLI signal was obtained as the difference from the response of the reference sensor (1 μg/ml S60-15+0 nM SORT1). A no-ligand control (0 μg/mL IgG+20 nM SORT1) demonstrated no measurable nonspecific antigen binding to the probe tip surface. Full and local fitting using a 1:1 interaction model was used to obtain association and dissociation rate constants (k _a and k _d , respectively) for each N33 mutant. Affinity constants (K _D ) were calculated as the ratio kd/ka. Various amino acid substitutions were made at position N33 and tested for binding to SORT1 by dissociation constant screening (under non-stress conditions) as indicated in Table. 9.

Table 9

Calculated ka, kd and K _D for binding of S-60-15 N33X mutants to SORT1

S-60-15 variants k _a (M c) ¹ k _d (c ¹ ) K _D (nM) S-60-15 [N33 (wild type (wt))] 5.12E+0 5 1.41E-04 270 S-60-15.1 [N33T] 4.7E+05 1.67E-04 344 S-60-15.2 [N33S] 5.63E+0 5 2.47E-04 440 S-60-15.3 [N33G] 4.98E+0 5 4.11E-04 825 S-60-15.4 [N33R] 5.24E+0 5 4.60E-04 880 S-60-15.5 [N33D] 5.90E+0 5 6.10E-04 1000 S-60-15.6 [N33H] 5.48E+0 5 4.60E-04 823 S-60-15.7 [N33K] 5.63E+0 5 5.47E-04 965 S-60-15.8 [N33Q] 6.09E+0 5 6.53E-04 905 S-60-15.9 [N33Y] 5.11E+0 5 6.75E-04 1285 S-60-15.10 [N33E] 6.76E+0 5 4.30E-03 2007 S-60-15.11 [N33W] 6.01E+0 5 5.79E-04 960 S-60-15.12 [N33F] 2.80E+0 8.16E-04 2910 5 S-60-15.13 [N33I] 4.91E+0 5 2.28E-04 460 S-60-15.14 [N33V] 4.45E+0 5 3.06E-04 690 S-60-15.15 [N33A] 4.93E+0 5 3.15E-04 625 S-60-15.16 [N33M] 4.95E+0 5 4.23E-04 850 S-60-15.17 [N33L] 4.12E+0 5 4.25E-04 1030

Note: ka, kd, and K _D represent averages across multiple experiments.

Screening of this panel of #3X mutants revealed that antibody S-60-15.5 (N33D), which mimics deamidation of N33, resulted in a significant reduction in binding, with a _KD of 1000 pM compared to 270 pM for wild-type N33 (wt ) (Table 9). In contrast, antibody S-60-15.1 (N33T) had the best affinity for retaining S-60-15 with SORT1, showing the smallest difference in K _D with

- 100 046146 wild type in the panel (Table 9).

In addition, a full kinetic analysis was performed using multiple concentrations of SORT1 for S-60-15.1 [N33T] in huIgG1 and huIgG1 LALAPS format in an independent experiment. Kinetic data are presented in table. 10. Kinetic data demonstrate that S60-15.1[N33T] is a high affinity antibody in both huIgG1 and huIgG1 LALAPS formats.

Table 10

Binding kinetic data, SORT1 for S-60-15.1 [N33T] in huIgG1 and huIgG 1 LALAP S format

Antibody designation k _a (M s) ¹ Me ¹ ) K _D (pm) S-60-15.1 [N33T] 5.99E+05 1.79E-04 298 S-60-15.1 [N33T] LALAPS 4.79E+05 1.27E-04 260

Cellular binding and functionality.

S-60-15.1 [N33T], in both huIgG1 and huIgG1 LALAPS formats, and S-6015.5 [N33D] were then tested for cell binding and functionality as described below.

Affinity matured antibodies S-60, S-60-15.1 [N33T], and S-60-15.5 [N33D] were tested in parallel for binding to human SORT1 expressed on HEK293T cells as described above. The results are presented in table. 11. S-60-15.1 [N33T] in huIgG1 or huIgG1 LALAPS format showed higher affinity than S-60 (Table 11), with a lower Kd and higher Bmax. The Fc variant did not appear to affect affinity, with only minor differences in KD and Bmax between S-60-15.1[N33T]huIgG1 and huIgG1 LALAPS. The fully deamidated variant S-60-15.5 [N33D] did not show any significant binding (Table 11, N.B. (no binding)) to cells expressing human SORT1, suggesting that amidation of the N33 residue is essential for target binding.

Table 11

Cell binding affinity values measured by FACS

Antibody designation ADnM) B _max (MFI) S60 1.02 758381 S-60-15.5 [N33D] N.B. N.B. S-60-15.1 [N33T] 0.3651 791996 S-60-15.1 [N33T] LALAPS 0.4231 792791

U251 cells, which endogenously express SORT1 and secrete PGRN, were incubated with the anti-SORT1 antibody S-60, S-60-15.1 [N33T] or S-60-15.5 [N33D] at different concentrations, and cell surface SORT1 levels were measured by FACS. as described in example 2. The control was untreated cells. A saturating concentration of 150 nM IgG antibody was used to quantify the percentage reduction in sortilin expression. A saturating concentration of 50 nM IgG antibody was used to quantify PGRN secretion. The results are shown in table. 12. S-60-15.1 [N33T] in huIgG1 or huIgG1 LALAPS format resulted in a significant decrease in SORT1 expression and an increase in PGRN secretion. Both the level of reduction in SORT1 expression and the level of increase in PGRN secretion were significantly improved for S-60-15.1[N33T] in huIgG1 or huIgG1 LALAPS format compared to S-60. S-60-15.1 [N33T] in huIgG1 format showed only a slight decrease in EC ₅₀ and percentage reduction in SORT1 expression, having a non-significant fold increase in PGRN secretion compared to S-60-15.1 [N33T] in huIgG1 LALAPS format. In contrast, the fully deamidated variant S-60-15.5 [N33D] had lower performance than S60 in each of these functional assays; it did not show a significant reduction in SORT1 expression and had only minimal changes in secreted PGRN levels.

- 101 046146

Table 12

Reduction of Cell Surface SORT1 Expression (DR) by S60 Antibody Variants

Antibody designation EC ₅₀ (hM) %DR at 150 nM antibody (IgG) Secretion of PGRN, fold compared to control at 50 nM antibody (IgG) S-60 680.9 62.5 1.75 S-60-15.5 [N33D] n.d. (undefined) 3.4 1.22 S-60-15.1 [N33T] 72.58 69.3 2.29 S-60-15.1 [N33T] LALAPS 103.6 70.3 1.97

The binding of biotinylated PGRN to SORT1 expressed on HEK293T cells was measured by FACS in the absence or presence of anti-SORT1 antibody S-60, S-60-15.1 [N33T] and S-6015.5 [N33D], as described in Example 3. Results are shown. in table 13. A saturating antibody concentration of 150 nM IgG was used to quantify the percentage of blocking of progranulin (PGRN) binding to SORT1.

Table 13

Blocking PGRN binding to SORT1 expressed on HEK293T cells

Antibody designation EC ₅₀ (hM) Blocking percentage at 150 nM antibody (IgG) S60 0.751 95.9 S-60-15.5 [N33D] 0.588 27.5 S-60-15.1 [N33T] 0.325 96.5 S-60-15.1 [N33T] LALAPS 0.405 96.9

The S-60-15.1 [N33T] variants, both with and without LALAPS, showed the lowest EC ₅₀ values and the highest level of blocking of PGRN binding to SORT1 compared to S-60-15.5 and also compared to S-60. Thus, S-60-15.1 [N33T] had better results than S-60 and S-60-15.5 in blocking PGRN binding to SORT1.

In general, S-60-15 was found to undergo significant deamidation at the N33 position under storage stress conditions (see Table 8), which increases the likelihood of degradation and leads to production implications. The previously described anti-SORT1 antibody S-60 and variants S-601, S-60-2, S-60-3, S-60-4, S-60-7 and S-60-8 also contain an asparagine at position 33 in VL. In particular, the position of the N33 deamidation site in the HVR-L1 site is likely to influence the binding of S-60-15 to SORT1. S-60-15.1 [N33T], one of several S-60-15 variants tested, retained significant binding affinity for SORT1 compared to S-60-15 (see Table 9) and showed significantly increased binding affinity for SORT1 by compared to S-60 (see Table 11; compare with S60-15 WT in Table 1). This retention of high binding affinity was unexpected for an antibody having a non-preferred substitution in HVR-L1, because residues of HVR-L1 can contact the antigen. Also unexpectedly, the S-60-15.1 [N33T] variant retained an increased degree of reduction in SORT1 expression, increased PGRN secretion, and increased blocking of progranulin (PGRN) binding to SORT1 compared to S-60 (see Tables 12 and 13). These results were also unexpected because a non-preferred substitution in the HVR sequence was expected to similarly negatively affect the functional characteristics of the antibody. Instead, S-60-15.1 [N33T] retains the desirable properties of reducing cell surface SORT1 expression, increasing PGRN secretion, and blocking PGRN binding to SORT1, in addition to being more stable than previously described antibodies due to the removal of sensitive manufacturing conditions of the remainder (manufacturing liability) in regulation 33 VL.

Example 5 Pharmacokinetics and pharmacodynamics of anti-sortilin antibody in non-human primates.

In this example, the pharmacokinetics (PK) and pharmacodynamics (PD) of intravenously (IV) administered anti-sortilin antibody S-60-15.1 [N33T]LALAPS were determined in non-human primates.

Materials and methods.

Single dose pharmacokinetic and pharmacodynamic studies.

For single-dose pharmacokinetic studies, cynomolgus monkeys were administered anti-sortilin antibody as a single intravenous (IV) dose of 5 mg/kg, 20 mg/kg, 60 mg/kg, or 200 mg/kg on Day 0 (n=3

- 102 046146 animals per dose). Blood and CSF were then collected from the animals at various time points to determine plasma and cerebrospinal fluid (CSF) anti-sortilin antibody concentrations required for anti-sortilin antibody pharmacokinetic measurements. Progranulin (PGRN) concentrations and sortilin (SORT1) levels on white blood cells (WBCs) required for pharmacodynamic measurements were also determined.

Anti-sortilin antibody concentrations were assessed using an ELISA assay with anti-sortilin antibody-specific anti-idiotypic antibodies. PGRN concentrations were assessed using a commercially available ELISA kit. SORT1 levels on leukocytes were analyzed using ELISA and normalized to protein concentration.

Results.

In table 14 shows the mean plasma Cmax, mean AUC (area under the curve, AUC), and t _1/2 for each of the anti-sortilin antibody doses tested.

Table 14

Cmax, mean AUC and t _1/2 for the indicated doses of anti-sortilin antibody (n=3 for each dose)

Antibody dose Average _Cmax (µg/ml) Average AUC (µg x h/ml) t%, hours 5 mg/kg 156 2,870 4.7 20 mg/kg 697 26,500 13.3 60 mg/kg 2,570 118,000 42 200 mg/kg 7,910 366,000 73.6

As shown in FIG. 1A, SORT1 expression levels in peripheral blood leukocytes were reduced following administration of any tested doses of anti-sortilin antibody to non-human primates. Higher doses of anti-sortilin antibody (60 mg/kg, 200 mg/kg) resulted in both an earlier and longer-lasting decrease in SORT1 levels in peripheral blood leukocytes compared with lower doses of anti-sortilin antibody (5 mg/kg, 20 mg/kg).

Plasma PGRN levels increased in non-human primates that received a single intravenous injection of anti-sortilin antibody in a time- and dose-dependent manner (Fig. 1B). Specifically, plasma PGRN levels increased 3- to 4-fold at Cmax compared to baseline levels for all anti-sortilin antibody doses tested. Plasma PGRN levels remained elevated for longer periods of time at higher doses of antibody. Additionally, increased plasma PGRN levels correlated with decreased SORT1 expression levels in peripheral blood leukocytes.

CSF levels of PGRN were also increased in non-human primates that received a single intravenous injection of anti-sortilin antibody. As shown in FIG. 1C, CSF PGRN levels increased 2- to 3-fold above baseline in animals receiving 20 mg/kg, 60 mg/kg, or 200 mg/kg. As with plasma PGRN levels, CSF PGRN levels remained elevated over time in the higher antibody dose groups.

In table 15 shows the mean CSF C _max , mean AUC, and t _1/2 for each of the anti-sortilin antibody doses tested in non-human primates. Concentrations of anti-sortilin antibody in CSF averaged about 0.1% of the amount observed in plasma.

Table 15

Pharmacokinetic parameters of anti-sortilin antibody in CSF and estimated __________ half-life in non-human primates__________

[1] Level doses Stach (MCG/ML) AUC _all (h*µg/ml) CL (ml/h/kg) th/2, hours (days) 5 mg/kg 20 184 20692 32.3 (1.34) 20 mg/kg 2243 35717 745 23.8(1) 60 mg/kg 6842 113573 623 38.3 (1.6) 200 mg/kg 4595 349187 1037 72.4 (3.02)

Pharmacokinetic and pharmacodynamic studies with repeated doses.

Additional pharmacokinetic and pharmacodynamic studies were conducted in non-human primates receiving anti-sortilin antibody on a repeat-dose schedule. In these studies, animals (2 males and 2 females) were administered anti-sortilin antibody at a dose of 60 mg/kg once a week for four weeks. SORT1 expression levels in peripheral blood leukocytes were then determined at various time points. In addition, we also determined

- 103 046146 plasma and CSF anti-sortilin antibody levels.

As shown in FIG. 2A, SORT1 levels in peripheral blood leukocytes remained reduced throughout the studies. Plasma PGRN levels increased at peak values 5-6 times higher than baseline levels (Fig. 2B). A decrease in plasma PGRN was observed after the fourth and final administration of anti-sortilin antibody; however, plasma PGRN levels remained elevated 2-fold over baseline. Additionally, CSF PGRN levels were elevated 3- to 4-fold from baseline (Fig. 2C).

Systemic exposure of anti-sortilin antibody, as assessed by mean Cmax and AUC _0-168 , was 2100 mcg/mL and 114,000 mcg/mLh on Day 1, and 3,020 mcg/mL and 174,000 mcg/mLH on Day 22. These results indicate that exposure on Day 22 was higher than on Day 1, indicating some accumulation of antibody.

The concentration of anti-sortilin antibody in the CSF of these animals ranged from 0.03% to 0.12% of that observed in plasma, which is consistent with the distribution of other antibodies in the CSF (Pestalozzi et al., (2000) J Clin Oncol 18( 11):2349-51; Petereit et al., (2009) Mult Scler 15(2):189-92).

Taken together, these results indicate that the S-60-15.1 [N33T] variant, despite its relatively short half-life, exhibits prolonged activity in vivo, reducing SORT1 levels in peripheral blood leukocytes and increasing plasma and CSF PGRN levels.

Additionally, intravenous (IV) administration of S-60-15.1 [N33T] to cynomolgus monkeys in repeated-dose toxicology studies of both 4 weeks and 26 weeks duration was well tolerated at doses up to 200 mg/kg weekly, and no adverse effects were observed. , which would preclude the possibility of conducting clinical studies in humans based on these and other toxicological studies that included assessment of cytokine release, mortality, body weight, respiratory rate and depth, and local tolerance at the injection site. Therefore, S-60-15.1 [N33T] is suitable for testing in clinical trials in people with FTD and other indications as specified herein.

The lack of adverse toxicity results with anti-sortilin antibody was surprising given that sortilin acts as a receptor for several ligands and has multiple functions in cellular transport and signaling (Nykjaer, A et al., (2012) Trends Neurosci 35: 261-270). It was expected that due to the variety of ligands and functional roles of sortilin, administration of an anti-sortilin antibody might cause undesirable effects; however, antibody S-60-15.1 [N33T] was well tolerated at doses administered.

Sequences of the VH regions of the S-60 antibody variants, alignment with S-60, and HVR positions.

S-60 (SEQ Ш NO:56)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-1 (SEQ ID NO:107)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSVRYWGWIRQPPGKGLEWIGSIYHSGSTY Y60

S-60-2 (SEQ ID NO:108)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSVRYWGWIRQPPGKGLEWIGAIYPSGST YY60

S-60-3 (SEQ ID NO:109)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST YY60

S-60-4 (SEQ ID NO:110)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-7 (SEQ ID NO: 111) QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST YY60 S-60-8 (SEQ ID NO: 112)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

- 104 046146

S-60-10 (SEQ ID NO:54)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-11 (SEQ ID NO:54)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST YY60

S-60-12 (SEQ ID NO:54)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-13 (SEQ ID NO:55)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-14 (SEQ ID NO:55)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-15 (SEQ ID NO:56)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-15.1 (SEQ ID NO:56)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-16 (SEQ ID NO:56)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-18 (SEQ ID NO:56)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-19 (SEQ ID NO:54)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60

S-60-24 (SEQ ID NO:56)

QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGST

YY60 ^^b1rs1VNIVs1NIE

S-60 HVR Hl (SEQ ID NO: 1)YSISSGYYWGS-60 HVR H2 (SEQ ID NO: 2)----------- ------------------------TIYHSGSTYY

S-60 (SEQ ID NO: 56 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT

VTVSS 122

S-60-1 (SEQ ID NO: 107 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-2 (SEQ ID NO: 108 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-3 (SEQ ID NO: 109 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT

- 105 046146

VTVSS 122

S-60-4 (SEQ ID NO: 110 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-7 (SEQ ID NO: 111 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-8 (SEQ ID NO: 112 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-10 (SEQ ID NO: 54 cont.)

NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTT VTVSS 122

S-60-11 (SEQ ID NO: 54 cont.)

NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTT VTVSS 122

S-60-12 (SEQ ID NO: 54 cont.)

NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTT VTVSS 122

S-60-13 (SEQ ID NO: 55 cont.)

NPSLESRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTT VTVSS 122

S-60-14 (SEQ ID NO: 55 cont.)

NPSLESRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTT VTVSS 122

S-60-15 (SEQ ID NO: 56 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-15.1 (SEQ ID NO: 56 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-16 (SEQ ID NO: 56 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-18 (SEQ ID NO: 56 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

S-60-19 (SEQ ID NO: 54 cont.)

NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTT VTVSS 122

S-60-24 (SEQ ID NO: 56 cont.)

NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTT VTVSS 122

Alignment

S-60 HVR H2 (SEQ ID NO: 2 cont.) NPSLKS-------------------------------------------------- ----------------S-60 HVR H3 (SEQ ID NO: 6)--------------------- --------------ARQGSIKQGYYGMDV-------- 106 046146

Sequences of the VL regions of the S-60 antibody variants, alignment with S-60, and HVR positions.

S-60 (SEQ ID NO: 79) DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60 S-60-1 (SEQ ID NO: 113)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-2 (SEQ ID NO:114)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-3 (SEQ ID NO:115)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQPPQLLIYLGSNR V60

S-60-4 (SEQ ID NO:116)

DIVMTQSPLSLPVTPGESASISCRSSQGLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-7 (SEQ ID NO:117)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-8 (SEQ ID NO:118)

- 107 046146

DIVMTQSPLSLPVTPGGPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-10 (SEQ ID NO:57)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-11 (SEQ ID NO:58)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-12 (SEQ ID NO:59)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQPPQLLIYLGSNR

V60

S-60-13 (SEQ ID NO:57)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-14 (SEQ ID NO:58)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-15 (SEQ ID NO:57)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-15.1 (SEQ Ш NO:60)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSTGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-16 (SEQ ID NO:77)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-18 (SEQ ID NO:78)

DIVMTQSPLSLPVTPGGPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-19 (SEQ ID NO:79)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

S-60-24 (SEQ ID NO:80)

DIVMTQSPLSLPVTPGESASISCRSSQGLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNR A 60

- 108 046146 βleveling

S-60 HVR LI (SEQ ID NO: 27)S-60 HVR L2 (SEQ ID NO: 29)RSSQSLLHSNGYNYLD------------------------- ------------------LGSNRA

S-60 (SEQ ID NO: 79 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-1 (SEQ ID NO: CONTINUED)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-2 (SEQ ID NO: 114 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-3 (SEQ ID NO: 115 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-4 (SEQ ID NO: 116 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-7 (SEQ ID NO: 117 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-8 (SEQ ID NO: 118 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-10 (SEQ ID NO: 57 cont.)

SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60-11 (SEQ ID NO: 58 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60-12 (SEQ ID NO: 59 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-13 (SEQ ID NO: 57 cont.)

SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60-14 (SEQ ID NO: 58 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60-15 (SEQ ID NO: 57 cont.)

SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60-15.1 (SEQ ID NO: 60 cont.)

SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60-16 (SEQ ID NO: 77 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGAYYCMQQQEAPLTFGGGTKVEIK 112

S-60-18 (SEQ ID NO: 78 cont.)

SGVPDRLSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-19 (SEQ ID NO: 79 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112

S-60-24 (SEQ ID NO: 80 cont.)

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112

S-60 Leveling

S-60 HVR L2 (SEQ ID NO: 29 cont.)8S-60 HVR L3 (SEQ ID NO: 33)------------MQQQETPLT- 109 046146

Table 16

Anti-SORTl antibody heavy chain H1 HVR sequences

Antibody(s) HVR Hl SEQ ID NO: S-60; S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S60-18; S-60-19; S-60-24 Table 17 Sequences of H2 HVR heavy chain an YSISSGYYWG [ti-SORTI anti 1 gel Antibody(s) HVRH2 SEQ ID NO: S-60; S-60-10; S-60-11; S-60-12; S-60-15 [N33 (wt)]; S-6015.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-6024 TIYHSGSTYYN PSLKS 2 S-60-13; S-60-14 TIYHSGSTYYN PSLES 3 Formula I TIYHSGSTYYN PSLXiS Xx represents K or E 4

Table 18

Anti-SORT1 antibody heavy chain H3 HVR sequences

Antibody(s) HVRH3 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-19 ARQGSIQQGYYGMD V 5 S-60; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-6015.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-24 ARQGSIKQGYYGMD V 6 Formula II ARQGSIXxQGYYGM D.V. Xx represents Q or K 7

- 110 046146

Table 19

L1 HVR light chain sequences of anti-SORTl antibodies

Antibody(s) HVR LI SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S60-14; S-60-15 [N33 (wt)]; S-60-16; S- RSSQSLLRSNGYNYLD 8 60-18 S-60-15.1 [N33T] RSSQSLLRSTGYNYLD 9 S-60-15.2 [N33S] RSSQSLLRSSGYNYLD 10 S-60-15.3 [N33G] RSSQSLLRSGGYNYLD AND S-60-15.4 [N33R] RSSQSLLRRSRGYNYLD 12 S-60-15.5 [N33D] RSSQSLLRSDGYNYLD 13 S-60-15.6 [N33H] RSSQSLLRSHGYNYLD 14 S-60-15.7 [N33K] RSSQSLLRSKGYNYLD 15 S-60-15.8 [N33Q] RSSQSLLRSQGYNYLD 16 S-60-15.9 [N33Y] RSSQSLLRSYGYNYLD 17 S-60-15.10 [N33E] RSSQSLLRSEGYNYLD 18 S-60-15.11 [N33W] RSSQSLLRSWGYNYLD 19 S-60-15.12 [N33F] RSSQSLLRSEGYNYLD 20 S-60-15.13 [N33I] RSSQSLLRSIGYNYLD 21 S-60-15.14 [N33V] RSSQSLLRSVGYNYLD 22 S-60-15.15 [N33A] RSSQSLLRSAGYNYLD 23 S-60-15.16 [N33M] RSSQSLLRSMGYNYLD 24 S-60-15.17 [N33L] RSSQSLLRSEGYNYLD 25 S-60; S-60-19 RSSQSLLHSNGYNYLD 26 S-60-24 RSSQGLLRSNGYNYLD 27 Formula III RSSQXALX2SX3GYNYLD Xi represents S or G X ₂ represents R or Η 28 Xs represents N, T, S, G, R, D, H, K, Q, Υ, E, W, F, I, V, A, M, or L

Table 20

L2 HVR light chain sequences of anti-SORT1 antibodies

Antibody(s) HVRL2 SEQ ID NO: S-60; S-60-10; S-60-11; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-6015.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60-24 LGSNRAS 29 S-60-12 LGSNRVS thirty Formula IV LGSNRXxS Xi is is A or V 31

- 111 046146

Table 21

L3 HVR light chain sequences of anti-SORTl antibodies

Antibody(s) HVRL3 SEQ ID NO: S-60-10; S-60-11; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-6015.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 MQQQEAPLT 32 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-24 S-60; S-60-12; S-60-18; S-60-19 MQQQETPLT 33 Formula V MQQQEX^L T Xi is is A or T 34

Table 22

Sequences of anti-SORT1 heavy chain framework region 1, antibodies

Antibody(s) VH ERI SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60-24 QVQLQESGPGLVKPSETLS LTCAVSG 35

Table 23

Anti-SORT1 heavy chain framework region 2 sequences. antibodies

Antibody(s) VHFR2 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-6015.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60-24 WIRQPPGK GLEWIG 36

- 112 046146

Table 24

Heavy chain framework region 3 sequences of anti-SORTl antibodies

Antibody(s) VH FR3 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-19 QVTISVDTSKNQFSLELSSVTA ADTAVYYC 37 S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-6015.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-24 RVTISVDTSKNQFSLKLSSVTA ADTAVYYC 38 Formula VI X ₁ VTISVDTSKNQFSLX ₂ LSSVT AADTAVYYC Xi represents Q or R X ₂ represents E or K 39

Table 25

Anti-SORT1 antibody heavy chain framework region 4 sequences

Antibody(s) VHFR4 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-6015.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60-24 WGQGTTV TVSS 40

- 113 046146

Table 26

Anti-SORTl light chain framework region 1 sequences

Antibody(s) VLFR1 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-15 DIVMTQ SPLSLPVTPG 41 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60- EPASISC 15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-6015.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-19 S-60-18 DIVMTQSPLSLPVTPG GPASISC 42 S-60-24 DIVMTQSPLSLPVTPG ESASISC DIVMTQSPLSLPVTPG X|X ₂ ASISC 43 Formula VII Xi represents E or G X ₂ represents P or S 44 Table 27 Sequences of the frame section 2 le! Antibody(s) S-60-10; S-60-11; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60- 15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S- which chain is anti-SORTl and VLFR2 Shtytel SEQ ID NO: 60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-6015.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-6019; S-60-24 WYLQKPGQSPQLLIY 45 S-60-12 WYLQKPGQPPQLLIY 46 WYLQKPGQXiPQLLIY Formula VIII Xi represents S 47 or P

- 114 046146

Table 28

Anti-SORTl light chain framework region 3 sequences

Antibody(s) VL FR3 SEQ ID NO: S-60-10; S-60-13; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-6015.4 [N33R]; S-60-15.5 [N33D]; S60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-6015.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-6015.16 [N33M]; S-60-15.17 [N33L] GVPDRF SSGSGSGTDFTLKISRAEAEDV GVYYC 48 S-60-11; S-60-12; S-60-14; S-60-19; S-60-24 GVPDRF SSGSGSGTDFTLKISRVEAEDV GVYYC 49 S-60-16 GVPDRF SSGSGSGTDFTLKISRVEAEDV GAYYC 50 S-60-18 GVPDRLSGSGSGTDFTLKISRVEAEDV GVYYC 51 Formula IX gvpdrx ₁ sgsgsgtdftlkisrx ₂ eaed VGX ₃ YYC Xi is F or L X ₂ is A or V X ₃ is V or A 52

Table 29

Anti-SORT1 antibody light chain framework region 4 sequences

Antibody(s) VLFR4 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-6015.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-6015.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; FGGGTKVEIK 53 S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60-24

- 115 046146

Table 30

Anti-SORTl antibody heavy chain variable region sequences

Antibody(s) HCVR SEQ ID NO: S-60-10, S-60-11, S-60-12, S-60-19 QVQLQESGPGLVKPSETLSLTCAVSGYSISSG YYWGWIRQPPGKGLEWIGTIYHSGSTYYNP SLKSQVTISVDTSKNQFSLELSSVTAADTAV YYC ARQGSIQQGY YGMD VWGQGTT VT VS S 54 S-30-13, S-60-14 QVQLQESGPGLVKPSETLSLTCAVSGYSISSG YYWGWIRQPPGKGLEWIGTIYHSGSTYYNP SLESRVTISVDTSKNQFSLKLSSVTAADTAV YYC ARQGSIQQGY YGMD VWGQGTT VT VS S 55 S-60, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-6015.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16, S-60-18, S-60-24 QVQLQESGPGLVKPSETLSLTCAVSGYSISSG YYWGWIRQPPGKGLEWIGTIYHSGSTYYNP SLKSRVTISVDTSKNQFSLKLSSVTAADTAV YYC ARQGSIKQGY YGMD VWGQGTT VT VS S 56

- 116 046146

Table 31

Anti-SORTl antibody light chain variable region sequences

Antibody(s) LCVR SEQ ID NO: S-60-10; S-60-13; S-60- 15 [N33 (wt)] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 57 S-60-11; S-60-14 DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRVEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 58 S-60-12 DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQPPQLLIYLGSNRVSGVP DRF SSGSGSGTDFTLKISRVEAED VGVYYCMQ QQETPLTFGGGTKVEIK 59 S-60-15.1 [N33T] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSTG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 60 S-60-15.2 [N33S] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSSG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 61 S-60-15.3 [N33G] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSG GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 62 S-60-15.4 [N33R] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSRG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 63 S-60-15.5 [N33D] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSD GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 64 S-60-15.6 [N33H] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSH GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP 65

- 117 046146

DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK S-60-15.7 [N33K] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSK GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SGSSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 66 S-60-15.8 [N33Q] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSQ GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SGSSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 67 S-60-15.9 [N33Y] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSY GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 68 S-60-15.10 [N33E] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSEG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 69 S-60-15.11 [N33W] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSW GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 70 S-60-15.12 [N33F] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSFG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 71 S-60-15.13 [N33I] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSIG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 72 S-60-15.14 [N33V] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSV GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 73

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S-60-15.15 [N33A] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSA GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 74 S-60-15.16 [N33M] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSM GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRAEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 75 S-60-15.17 [N33L] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSLG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIK 76 S-60-16 DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRVEAED VGAYYCMQ QQEAPLTFGGGTKVEIK 77 S-60-18 DIVMTQSPLSLPVTPGGPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRLSGSGSGTDFTLKISRVEAEDVGVYYCMQ QQETPLTFGGGTKVEIK 78 S-60, S-60-19 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRVEAED VGVYYCMQ QQETPLTFGGGTKVEIK 79 S-60-24 DIVMTQSPLSLPVTPGESASISCRSSQGLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRF SSGSGSGTDFTLKISRVEAED VGVYYCMQ QQEAPLTFGGGTKVEIK 80

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Table 32

Amino acid sequences of sortilin

Description Subsequence SEQ ID NO Human sortilin 10 20 30 40 50 MERPWGAADG LSRWPHGLGL LLLLQLLPPS TLSQDRLDAP PPPAAPLPRW 60 70 80 90 100 SGPIGVSWGL RAAAAGGAFP RGGRWRRSAP GEDEECGRVR DFVAKLANNT 110 120 130 140 150 IIQIIVFDDLRG SVSLSWVGDS TGVILVLTTF IIVPLVIMTFG QSKLYRSEDY 160 170 180 190 200 GKNFKDITDL INNTFIRTEF GMAIGPENSG KVVLTAEVSG GSRGGR1FRS 210 220 230 240 250 SDFAKNFVQT DLPFHPLTQM MYSPQNSDYL LALSTENGLW VSKNFGGKWE 260 270 280 290 300 EIHKAVCLAK WGSDNTIFFT TYANGSCKAD LGALELWRTS DLGKSFKTIG 310 320 330 340 350 VKIYSFGI.GG RFLFASVMAD KDTTRRIHVS TDQGDTWSMA QLPSVGQEQF 360 370 380 390 400 YSILAANDDM VFMHVDEPGD TGFGTIFTSD DRGIVYSKSL DRHT.YTTTGG 410 420 430 440 450 ETDFTNVTSL RGVYITSVLS EDNSIQTMIT FDQGGRWTHL RKPENSECDA 460 470 480 490 500 TAKNKNECSL H1HASYS1SQ KLNVPMAPES EPNAVG1V1A HGSVGDAISV 510 520 530 540 550 MVPDVY1SDD GGYSWTKMLE GPHYY'HLDS GG11VA1EHS SRPINVIKFS 560 570 580 590 600 TDEGQCWQTY 1’FTRDPIYFT GLASEPGARS MN1S1WGF1E SFLTSQWVSY 81

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Mouse sortilin Sortilip rat 610 620 630 640 650 TIDFKDTLER NCEEKDYTIW LAHSTDPEDY EDGCTLGYKE QFLRLRKSSV 660 670 680 690 700 CQNGRDYVVT KQPS1CLCSL EDFLCDFGYY RPENDSKCVE QPELKGHDLE 710 720 730 740 750 FCLYGREEHL TTNGYRKIPG DKCQGGVNPV REVKDLKKKC TSNFLSPEKQ 760 770 780 790 800 NSKSNSVPII LAIVGLMLVT VVAGVLIVKK YVCGGRFLVH RYSVLQQHAE 810 820 830 ANGVDGVDAL DTASHTNKSG YHDDSDEDLL E MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG PVGVSWGLRA AAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HVFDDLSGSV SLSWVGDSTG VILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGM AIGPENSGKV ILTAEVSGGS RGGR VFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLA LSTENGLWVS KNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDL GKTFKTIGVK IYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYS ILAANEDMVF MHVDEPGDTG FGTIFTSD DR GIVYSKSLDR HLYTTTGGET DFTNVTSLRG VYITSTLSED NSIQSMITFD QGGRWEHLRK PENSKCDATA KNKNECSLHI HASYSISQKL NVPMAPLSEP NAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMIEGP HYYTILDSGG IIVAIEHSNR PINVKFSTD EGQCWQSYVF TQEPIYFTGL ASEPGARSMN ISIWGFTESF ITRQWVSYTV DFKDILERNC EEDDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQ NGRDYVVAKQ PSVCPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTT NGYRKIPGDK CQGGMNPARE VKDLKK KCTS NFLNPTKQNS KSNSVPIILA IVGLMLVTW AGVLIVKKYV CGGRFLVHRY SVLQQHAEAD GVEALDSTSH AKSG YHDDSE EDLEE MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG PVGVSWGLRA AAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HV FDDLSGSV SLSWVGDSTG VILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGM AIGPENSGKV ILTAEVSGGS RCiGRVFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLA LSTENGLWVS KNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDL GKTFKTIGVK IYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYS ILAANDDMVF MHVDEPGDTG FGTIFTSDDR GIVYSKSLDR HLYTTTGGET DFTNV TSLRG VYITSTLSED NSIQSMITFD QGGRWEHLQK PENSKCDATA KNKNECSLHI HASYSISQKL NVPMAPLSEP NAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMLEGP HYYTILDSGG IIVAIEHSNR PINVKFSTD EGQCWQSYVF SQEPVYFTGL ASEPGARSMN ISIWGFTESF LTRQWVSY H DFKDILERNC EENDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQ NGRDYVVAKQ PSICPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTT NGYRKIPGDR CQGGMNPARE VKDLKKKCTS NFLNPKFCQNS KSSSVPIILA IVGLMLVTW AGVLIVKKYV CGGRFLVHRY SVLQQHAEAD GVEALDTASH AKSGYHDDSD EDLLE 82 83

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Table 33

Peptide sequences S-60-15

Description Subsequence SEQ ID NO Asp-box motif (S/T)-X-(D/N)-X-X-X-X-(W/F/Y) X represents any amino acid 84 Asp-box motif X-X-(S/T)-X-(D/N)-X-G-X-(T/S)-(W/F/Y)-X X represents any amino acid 85 Asp box motif in human sortilin (residues 200-207) SSDFAKNF 86 Peptide S-60-15 NQFSLK 87 Peptide S-60-15 QGYYGMD VWGQGTTVT VS S ASTK 88 Peptide S-60-15 DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTK 89 Peptide S-60-15 DTLMISR 90 Peptide S-60-15 TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHN A K 91 Peptide S-60-15 VVSVLTVLHQDWLNGK 92 Peptide S-60-15 EPQVYTLPPSQEEMTK 93 Peptide S-60-15 NQVSLTCLVK 94 Peptide S-60-15 GFYPSDIAVEWESNGQPENNYK 95 Peptide S-60-15 WQEGNVF SC S VMHE ALHNHYTQK 96 Peptide S-60-15 DIVMTQSPLSLPVTPGEPASISCR 97 Peptide S-60-15 SSQSLLR 98 Peptide S-60-15 SNGYNYLDWYLQKPGQSPQLLIYLGSNR 99 Peptide S-60-15 AEAEDVGVYYCMQQQEAPLTFGGGTK 100 Peptide S-60-15 SGTASVVCLLNNFYPR 101 Peptide S-60-15 VDNALQSGNSQESVTEQDSK 102

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Table 34

Amino acid sequences of the Fc domain

Description Subsequence SEQ ID NO huIgGl LALAPSFc ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVL TVLHQ DWLNGKEYKCKVSNKALPASIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK 103 huIgGl LALAPS Fc without C- terminal lysine ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVL TVLHQ DWLNGKEYKCKVSNKALPASIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPG 104

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Table 35

Amino acid sequences of the full-length heavy chain

Description S-60-10, S-60- Subsequence QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWI SEQ ID NO 105 11, S-60-12, S60-19 with__Fc LALAPS with Scone Lysine RQPPGKGLEWIGTIYHSGSTYYNPSLKSQVTISVDTSKN QF SLELS SVTAADTAVYYCARQGSIQQGYYGMDVWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVI<FNWYVDGVEVHNAI<TI<PREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S-60-10, S-6011, S-60-12, S60-19 with__Fc LALAPS without C-terminal lysine QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWI RQPPGKGLEWIGTIYHSGSTYYNPSLKSQVTISVDTSKN QF SLELS SVT AADTAVYYCARQGSIQQGYYGMD VWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVT VPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK GF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 106 S-60-13, S-6014 with Fc LALAPS with Scone Lysine QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWI RQPPGKGLEWIGTIYHSGSTYYNPSLESRVTISVDTSKN QF SLKL S SVT AADTAVYYCARQGSIQQGYYGMD VWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVI<FNWYVDGVEVHNAI<TI<PREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSL TCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 135

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S-60-13, S-6014 with Fc LALAPS without C-terminal lysine QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWI RQPPGKGLEWIGTIYHSGSTYYNPSLESRVTISVDTSKN QF SLKL S S VT AADT AVYYC ARQGSIQQGYYGMD VWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVI<FNWYVDGVEVHNAI<TI<PREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK TISKAKGQPREPQVYTLPPSRDELTKNQ VSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 136 S-60, S-60-15 [N33 (wt)], S60-15.1 [N33T], S-6015.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60- 15.5 [N33D], S-6015.6 [N33H], S-60-15.7 [N33K], S-6015.8 [N33Q], S-60-15.9 [N33Y], S-6015.10 [N33E], S-60-15.11 [ N33W], S-6015.12 [N33F], S-60-15.13 [N33I], S-6015.14 [N33V], QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWI RQPPGKGLEWIGTIYHSGSTYYNPSLKSRVTISVDTSKN QF SLKL S SVT AADT AVYYC ARQGSIKQGYYGMD VWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVI<FNWYVDGVEVHNAI<TI<PREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK TISKAKGQPREPQVYTLPPSRDELTKNQV SLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 137

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S-60-15.15 [N33A], S-6015.16 [N33M], S-60-15.17 [N33L], S-6016, S-60-18, S- 60-24 c__Fc LALAPS with C-terminal lysine S-60, S-60-15 [N33 (wt)], S60-15.1 [N33T], S-6015.2 [N33S], S-60-15.3 [N33G], S-6015.4 [N33R], S-60- 15.5 [N33D], S-6015.6 [N33H], S-60-15.7 [N33K], S-6015.8 [N33Q], S-60-15.9 [N33Y], S-6015.10 [N33E], S-60-15.11 [ N33W], S-6015.12 [N33F], S-60-15.13 [N33I], S-6015.14 [N33V], S-60-15.15 [N33A], S-60- QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWI RQPPGKGLEWIGTIYHSGSTYYNPSLKSRVTISVDTSKN QF SLKL S S VT AADT AVYYC ARQGSIKQGYYGMD VWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVI<FNWYVDGVEVHNAI<TI<PREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK TISKAKGQPREPQVYTLPPSRDELTKNQ VSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 138 15.16 [N33M], S-60-15.17 [N33L], S-6016, S-60-18, S60-24 c__Fc LALAPS without C-terminal lysine

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Table 36

Amino acid sequences of the full-length light chain

Description Subsequence SEQ ID NO S-60-10; S60-13; S-6015 [N33 (wt)] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 139 S-60-11; S60-14 DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YE KHKVYACEVTHQGLSSPVTKSFNRGEC 140 S-60-12 DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLDWYLQKPGQPPQLLIYLGSNRVSGVP DRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ QQETPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YE KHKVYACEVTHQGLSSPVTKSFNRGEC 141 S-60-15.1 [N33T] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSTG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQ 142

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L1<SGTASVVCLLNNFYPREAI<VQWI<VDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.2 [N33S] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSSG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQ L1<SGTASVVCLLNNFYPREAI<VQWI<VDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC 143 S-60-15.3 [N33G] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSG GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 144 S-60-15.4 [N33R] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSR GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 145 S-60-15.5 [N33D] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSD GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 146 S-60-15.6 [N33H] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSH GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE 119

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QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.7 [N33K] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSK GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YE KHKVYACEVTHQGLSSPVTKSFNRGEC 120 S-60-15.8 [N33Q] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSQ GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YE KHKVYACEVTHQGLSSPVTKSFNRGEC 121 S-60-15.9 [N33Y] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSY GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 122 S-60-15.10 [N33E] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSEG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQ L1<SGTASVVCLLNNFYPREAI<VQWI<VDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC 123 S-60-15.11 [N33W] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSW GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE 124

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QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.12 [N33F] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSFG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQ L1<SGTASVVCLLNNFYPREAI<VQWI<VDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC 125 S-60-15.13 [N33I] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSIG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQ L1<SGTASVVCLLNNFYPREAI<VQWI<VDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC 126 S-60-15.14 [N33V] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSV GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 127 S-60-15.15 [N33A] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSA GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEK HKVYACEVTHQGLSSPVTKSFNRGEC 128 S-60-15.16 [N33M] DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSM GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQ QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE 129

- 130 046146

QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSLG YNYLDWYLQKPGQSPQLLIYLGSNRASGVPD S-60-15.17 [N33L] RFSGSGSGTDFTLKISRAEAEDVGVYYCMQQ QEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQ L1<SGTASVVCLLNNFYPREAI<VQWI<VDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSN GYNYLD WYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRVEAEDVGAYYCMQ 130 S-60-16 QQEAPLTFGGGTKVEIKRT VAAP S VFIFPP SDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC DIVMTQSPLSLPVTPGGPASISCRSSQSLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRLSGSGSGTDFTLKISRVEAEDVGVYYCMQ 131 S-60-18 QQETPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP 132 S-60, S-60- Ί 0 DRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ QQETPLTFGGGTKVEIKRTVAAPSVFIFPPSDE 133 1 27 QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC DIVMTQSPLSLPVTPGESASISCRSSQGLLRSN GYNYLDWYLQKPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ S-60-24 QQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC 134

- 131 046146

List of G sequences <110> Alector LLC <120> ANTIBODIES AGAINST SORTILIN AND METHODS OF THEIR APPLICATION <130> 73502-20019.41 <140> Not assigned yet <141> Simultaneously with this document <150> US 62/868,849 <151> 2019-06- 28 <150> US 62/860,184 <151> 2019-06-11 <150> US 62/698,007 <151> 2018-07-13 <160> 146 <170> FastSEQ for Windows version 4.0 <210> 1 <211> 10 < 212> PROTEIN < 213> Artificial sequence < 220>

<223> Synthetic construct <400> 1

Tyr Ser Ile Ser Ser Gly Tyr Tyr Trp Gly

5 10 < 210> 2 < 211> 16 < 212> PROTEIN < 213> Artificial sequence < 220>

<223> Synthetic construct <400> 2

Thr Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser

5 10 15 < 210> 3 < 211> 16 < 212> PROTEIN < 213> Artificial sequence < 220>

<223> Synthetic construct <400> 3

Thr Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Glu Ser

5 10 15

- 132 046146 <210> 4 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 15 <223> Xaa = Lys or Glu <400> 4

Thr Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Xaa Ser

5 10 15 <210> 5 <211> 15 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 5

Ala Arg Gln Gly Ser Ile Gln Gln Gly Tyr Tyr Gly Met Asp Val 1 5 10 15 <210> 6 <211> 15 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 6

Ala Arg Gln Gly Ser Ile Lys Gln Gly Tyr Tyr Gly Met Asp Val 1 5 10 15 <210> 7 <211> 15 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 7 <223> Xaa = Gln or Lys <400> 7

Ala Arg Gln Gly Ser Ile Xaa Gln Gly Tyr Tyr Gly Met Asp Val 1 5 10 15 <210> 8

- 133 046146 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 8

Arg Ser Ser Gln Ser Leu Leu Arg Ser Asn Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 9 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 9

Arg Ser Ser Gln Ser Leu Leu Arg Ser Thr Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 10 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 10

Arg Ser Ser Gln Ser Leu Leu Arg Ser Ser Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 11 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 11

Arg Ser Ser Gln Ser Leu Leu Arg Ser Gly Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 12 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 12

Arg Ser Ser Gln Ser Leu Leu Arg Ser Arg Gly Tyr Asn Tyr Leu Asp

5 10 15

- 134 046146 <210> 13 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 13

Arg Ser Ser Gln Ser Leu Leu Arg Ser Asp Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 14 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 14

Arg Ser Ser Gln Ser Leu Leu Arg Ser His Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 15 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 15

Arg Ser Ser Gln Ser Leu Leu Arg Ser Lys Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 16 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 16

Arg Ser Ser Gln Ser Leu Leu Arg Ser Gln Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 17 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 17

Arg Ser Ser Gln Ser Leu Leu Arg Ser Tyr Gly Tyr Asn Tyr Leu Asp

- 135 046146

5 10 15 <210> 18 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 18

Arg Ser Ser Gln Ser Leu Leu Arg Ser Glu Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 19 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 19

Arg Ser Ser Gln Ser Leu Leu Arg Ser Trp Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 20 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 20

Arg Ser Ser Gln Ser Leu Leu Arg Ser Phe Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 21 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 21

Arg Ser Ser Gln Ser Leu Leu Arg Ser Ile Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 22 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct

- 136 046146 <400> 22

Arg Ser Ser Gln Ser Leu Leu Arg Ser Val Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 23 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 23

Arg Ser Ser Gln Ser Leu Leu Arg Ser Ala Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 24 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 24

Arg Ser Ser Gln Ser Leu Leu Arg Ser Met Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 25 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 25

Arg Ser Ser Gln Ser Leu Leu Arg Ser Leu Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 26 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 26

Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 27 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

- 137 046146 <223> Synthetic construct <400> 27

Arg Ser Ser Gln Gly Leu Leu Arg Ser Asn Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 28 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 5 <223> Xaa = Ser or Gly <220>

<221> VARIANT <222> 8 <223> Xaa = Arg or His <220>

<221> VARIANT <222> 10 <223> Xaa = Asn, Thr, Ser, Gly, Arg, Asp, His, Lys, Gln, Tyr, Glu, Trp, Phe, Ile, Val, Ala, Met, or Leu < 400>28

Arg Ser Ser Gln Xaa Leu Leu Xaa Ser Xaa Gly Tyr Asn Tyr Leu Asp

5 10 15 <210> 29 <211> 7 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 29

Leu Gly Ser Asn Arg Ala Ser

5 <210> 30 <211> 7 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 30

Leu Gly Ser Asn Arg Val Ser

5 <210> 31

- 138 046146 <211> 7 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

< 221> VARIANT < 222> 6 < 223> Xaa = Ala or Val < 400> 31

Leu Gly Ser Asn Arg Xaa Ser

5 <210> 32 <211> 9 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 32

Met Gln Gln Gln Glu Ala Pro Leu Thr

5 <210> 33 <211> 9 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 33

Met Gln Gln Gln Glu Thr Pro Leu Thr

5 <210> 34 <211> 9 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 6 <223> Xaa = Ala or Thr <400> 34

Met Gln Gln Gln Glu Xaa Pro Leu Thr

5 <210> 35 <211> 26

- 139 046146 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 35

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser 1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly

25 <210> 36 <211> 14 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 36

Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly

5 10 <210> 37 <211> 30 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 37

Gln Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 1 5 10 15

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 20 25 30 <210> 38 <211> 30 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 38

Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 1 5 10 15

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 20 25 30

Glu

Glu

Lys <210> 39 <211> 30 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct

- 140 046146 <220>

<221> VARIANT <222> 1 <223> Xaa = Gln or Arg <220>

<221> VARIANT <222> 16 <223> Xaa = Glu or Lys <400> 39

Xaa Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 1 5 10 15 Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 20 25 thirty

Xaa <210> 40 <211> 11 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 40

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

5 10 <210> 41 <211> 23 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 41

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro 1 5 10 15

Glu Pro Ala Ser Ile Ser Cys 20 <210> 42 <211> 23 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 42

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro 1 5 10 15

Gly Pro Ala Ser Ile Ser Cys 20

Gly

Gly <210> 43 <211> 23

- 141 046146 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 43

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15

Glu Ser Ala Ser Ile Ser Cys <210> 44 <211> 23 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 17 <223> Xaa = Glu or Gly <220>

<221> VARIANT <222> 18 <223> Xaa = Pro or Ser <400> 44

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15

Xaa Xaa Ala Ser Ile Ser Cys <210> 45 <211> 15 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 45

Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr 1 5 10 15 <210> 46 <211> 15 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 46

Trp Tyr Leu Gln Lys Pro Gly Gln Pro Pro Gln Leu Leu Ile Tyr 1 5 10 15

- 142 046146 <210> 47 <211> 15 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 9 <223> Xaa = Ser or Pro <400> 47

Trp Tyr Leu Gln Lys Pro Gly Gln Xaa Pro Gln Leu Leu Ile Tyr 1 5 10 15 <210> 48 <211> 32 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 48

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Lys Ile Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys 20 25 thirty

<210> 49 <211> 32 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 49

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15

Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys 20 25 30 <210> 50 <211> 32 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 50

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15

Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Ala Tyr Tyr Cys 20 25 30

- 143 046146 <210> 51 <211> 32 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 51

Gly Val Pro Asp Arg Leu Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15

Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys 20 25 30 <210> 52 <211> 32 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 6 <223> Xaa = Phe or Leu <220>

<221> VARIANT <222> 22 <223> Xaa = Ala or Val <220>

<221> VARIANT <222> 29 <223> Xaa = Val or Ala <400> 52

Gly Val Pro Asp Arg Xaa Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Lys Ile Ser Arg Xaa Glu Ala Glu Asp Val Gly Xaa Tyr Tyr Cys 20 25 thirty

<210> 53 <211> 10 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 53

Phe Gly Gly Gly Thr

5 sequence construct

Lys Val Glu Ile Lys <210> 54 <211> 122 <212> PROTEIN <213> Artificial sequence

- 144 046146 <220>

<223> Synthetic construct <400> 54

Gln Val Gln Leu Gln Glu Ser Gly 15

Thr Leu Ser Leu Thr Cys Ala Val 20

Tyr Tyr Trp Gly Trp Ile Arg Gln 3540

Ile Gly Thr Ile Tyr His Ser Gly 5055

Lys Ser Gln Val Thr Ile Ser Val 6570

Leu Glu Leu Ser Ser Val Thr Ala 85

Ala Arg Gln Gly Ser Ile Gln Gln 100

Gly Gln Gly Thr Thr Val Thr Val

115120

Pro Gly Leu Val Lys Pro Ser Glu 1015

Ser Gly Tyr Ser Ile Ser Ser Gly 2530

Pro Pro Gly Lys Gly Leu Glu Trp 45

Ser Thr Tyr Tyr Asn Pro Ser Leu 60

Asp Thr Ser Lys Asn Gln Phe Ser 7580

Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Gly Tyr Tyr Gly Met Asp Val Trp 105110

Ser Ser <210> 55 <211> 122 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 55

Gln Val Gln Leu Gln Glu Ser Gly 15

Thr Leu Ser Leu Thr Cys Ala Val 20

Tyr Tyr Trp Gly Trp Ile Arg Gln 3540

Ile Gly Thr Ile Tyr His Ser Gly 5055

Glu Ser Arg Val Thr Ile Ser Val 6570

Leu Lys Leu Ser Ser Val Thr Ala 85

Ala Arg Gln Gly Ser Ile Gln Gln 100

Gly Gln Gly Thr Thr Val Thr Val

115120

Pro Gly Leu Val Lys Pro Ser Glu 1015

Ser Gly Tyr Ser Ile Ser Ser Gly 2530

Pro Pro Gly Lys Gly Leu Glu Trp 45

Ser Thr Tyr Tyr Asn Pro Ser Leu 60

Asp Thr Ser Lys Asn Gln Phe Ser 7580

Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Gly Tyr Tyr Gly Met Asp Val Trp 105110

Ser Ser <210> 56 <211> 122 <212> PROTEIN <213> Artificial sequence <220>

<223>

<400> 56

Gln Val Gln Leu 1

Thr Leu Ser Leu

Synthetic construct

Gln Glu Ser Gly 5

Thr Cys Ala Val

Pro Gly Leu Val 10

Ser Gly Tyr Ser 25

Lys Pro Ser Glu 15

Ile Ser Ser Gly 30

- 145 046146

Tyr Tyr Trp Gly 35

Ile Gly Thr Ile

Lys Ser Arg Val 65

Leu Lys Leu Ser

Ala Arg Gln Gly 100

Gly Gln Gly Thr 115

Trp Ile Arg Gln Pro Pro 40

Tyr His Ser Gly Ser Thr 55

Thr Ile Ser Val Asp Thr 70

Ser Val Thr Ala Ala Asp 85 90

Ser Ile Lys Gln Gly Tyr 105

Thr Val Thr Val Ser Ser 120

Gly Lys Gly Leu Glu Trp 45

Tyr Tyr Asn Pro Ser Leu 60

Ser Lys Asn Gln Phe Ser 75 80

Thr Ala Val Tyr Tyr Cys 95

Tyr Gly Met Asp Val Trp 110 <210> 57 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 57

Asp Ile Val Met 1

Glu Pro Ala Ser

Asn Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro

100

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110 <210> 58 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 58

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110

- 146 046146 <210> 59 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 59

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Pro 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Val Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 60 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 60

Asp Ile Val Met Thr Gln Ser Pro 15

Glu Pro Ala Ser Ile Ser Cys Arg 20

Thr Gly Tyr Asn Tyr Leu Asp Trp 3540

Pro Gln Leu Leu Ile Tyr Leu Gly 5055

Asp Arg Phe Ser Gly Ser Gly Ser 6570

Ser Arg Ala Glu Ala Glu Asp Val 85

Gln Glu Ala Pro Leu Thr Phe Gly 100

Leu Ser Leu Pro Val Thr Pro Gly 10 15

Ser Ser Gln Ser Leu Leu Arg Ser 25 30

Tyr Leu Gln Lys Pro Gly Gln Ser 45

Ser Asn Arg Ala Ser Gly Val Pro 60

Gly Thr Asp Phe Thr Leu Lys Ile 7580

Gly Val Tyr Tyr Cys Met Gln Gln 9095

Gly Gly Thr Lys Val Glu Ile Lys 105110 <210> 61 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 61

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser

- 147 046146

Ser Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro

100

2530

Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln 4045

Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val 5560

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys 7075

Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln 85 9095

Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 105110

Ser

Pro

Ile 80 Gln

Lys <210> 62 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 62

Asp Ile Val Met Thr Gln Ser Pro 15

Glu Pro Ala Ser Ile Ser Cys Arg 20

Gly Gly Tyr Asn Tyr Leu Asp Trp 3540

Pro Gln Leu Leu Ile Tyr Leu Gly 5055

Asp Arg Phe Ser Gly Ser Gly Ser 6570

Ser Arg Ala Glu Ala Glu Asp Val 85

Gln Glu Ala Pro Leu Thr Phe Gly 100

Leu Ser Leu Pro Val Thr Pro Gly 10 15

Ser Ser Gln Ser Leu Leu Arg Ser 25 30

Tyr Leu Gln Lys Pro Gly Gln Ser 45

Ser Asn Arg Ala Ser Gly Val Pro 60

Gly Thr Asp Phe Thr Leu Lys Ile 7580

Gly Val Tyr Tyr Cys Met Gln Gln 9095

Gly Gly Thr Lys Val Glu Ile Lys 105110 <210> 63 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 63

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Arg Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110

- 148 046146 <210> 64 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 64

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Asp Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 65 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 65

Asp Ile Val Met 1

Glu Pro Ala Ser 20

His Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro 100

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110 <210> 66 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223>

<400> 66

Asp Ile Val Met 1

Glu Pro Ala Ser

Synthetic construct

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

- 149 046146

Lys Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro

100

Tyr Leu Asp Trp Tyr Leu 40

Ile Tyr Leu Gly Ser Asn 55

Gly Ser Gly Ser Gly Thr 70

Ala Glu Asp Val Gly Val 85 90

Leu Thr Phe Gly Gly Gly 105

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile

110

Ser

Pro

Ile 80 Gln

Lys <210> 67 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 67

Asp Ile Val Met Thr Gln Ser Pro 15

Glu Pro Ala Ser Ile Ser Cys Arg 20

Gln Gly Tyr Asn Tyr Leu Asp Trp 3540

Pro Gln Leu Leu Ile Tyr Leu Gly 5055

Asp Arg Phe Ser Gly Ser Gly Ser 6570

Ser Arg Ala Glu Ala Glu Asp Val 85

Gln Glu Ala Pro Leu Thr Phe Gly 100

Leu Ser Leu Pro Val Thr Pro Gly 10 15

Ser Ser Gln Ser Leu Leu Arg Ser 25 30

Tyr Leu Gln Lys Pro Gly Gln Ser 45

Ser Asn Arg Ala Ser Gly Val Pro 60

Gly Thr Asp Phe Thr Leu Lys Ile 7580

Gly Val Tyr Tyr Cys Met Gln Gln 9095

Gly Gly Thr Lys Val Glu Ile Lys 105110 <210> 68 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 68

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Tyr Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 69

- 150 046146 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 69

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Glu Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 70 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 70

Asp Ile Val Met 1

Glu Pro Ala Ser

Trp Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro

100

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110 <210> 71 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 71

Asp Ile Val Met 1

Glu Pro Ala Ser 20

Phe Gly Tyr Asn

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Tyr Leu Asp Trp

Leu Ser Leu Pro Val Thr Pro Gly

15

Ser Ser Gln Ser Leu Leu Arg Ser

thirty

Tyr Leu Gln Lys Pro Gly Gln Ser

- 151 046146

Pro Gln Leu 50

Asp Arg Phe 65

Ser Arg Ala

Gln Glu Ala

4045

Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 5560

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 70 7580

Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gln 85 9095

Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 72 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 72

Asp Ile Val Met Thr Gln Ser Pro 15

Glu Pro Ala Ser Ile Ser Cys Arg 20

Ile Gly Tyr Asn Tyr Leu Asp Trp 3540

Pro Gln Leu Leu Ile Tyr Leu Gly 5055

Asp Arg Phe Ser Gly Ser Gly Ser 6570

Ser Arg Ala Glu Ala Glu Asp Val 85

Gln Glu Ala Pro Leu Thr Phe Gly 100

Leu Ser Leu Pro Val Thr Pro Gly 10 15

Ser Ser Gln Ser Leu Leu Arg Ser 25 30

Tyr Leu Gln Lys Pro Gly Gln Ser 45

Ser Asn Arg Ala Ser Gly Val Pro 60

Gly Thr Asp Phe Thr Leu Lys Ile 7580

Gly Val Tyr Tyr Cys Met Gln Gln 9095

Gly Gly Thr Lys Val Glu Ile Lys 105110 <210> 73 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 73

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Val Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 74 <211> 112

- 152 046146 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 74

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Ala Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110 <210> 75 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 75

Asp Ile Val Met Thr Gln Ser Pro 15

Glu Pro Ala Ser Ile Ser Cys Arg 20

Met Gly Tyr Asn Tyr Leu Asp Trp 3540

Pro Gln Leu Leu Ile Tyr Leu Gly 5055

Asp Arg Phe Ser Gly Ser Gly Ser 6570

Ser Arg Ala Glu Ala Glu Asp Val 85

Gln Glu Ala Pro Leu Thr Phe Gly 100

Leu Ser Leu Pro Val Thr Pro Gly 10 15

Ser Ser Gln Ser Leu Leu Arg Ser 25 30

Tyr Leu Gln Lys Pro Gly Gln Ser 45

Ser Asn Arg Ala Ser Gly Val Pro 60

Gly Thr Asp Phe Thr Leu Lys Ile 7580

Gly Val Tyr Tyr Cys Met Gln Gln 9095

Gly Gly Thr Lys Val Glu Ile Lys 105110 <210> 76 <211> 112 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 76

Asp Ile Val Met 1

Glu Pro Ala Ser

Leu Gly Tyr Asn 35

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Tyr Leu Asp Trp 40

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

- 153 046146

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu

100

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys 7075

Glu Asp Val Gly Val Tyr Tyr Cys Met Gln 9095

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

105110

Pro

Ile 80 Gln

Lys <210> 77 <211> 112 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 77

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Ala Pro Leu

100 sequence construct

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln 4045

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys 7075

Glu Asp Val Gly Ala Tyr Tyr Cys Met Gln 9095

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 105110

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys <210> 78 <211> 112 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 78

Asp Ile Val Met Thr 1 5

Gly Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Leu Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Thr Pro Leu

100 sequence construct

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln 4045

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys 7075

Glu Asp Val Gly Val Tyr Tyr Cys Met Gln 9095

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 105110

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys <210> 79 <211> 112 <212> PROTEIN

- 154 046146 <213> Artificial sequence <220>

<223> Synthetic construct <400> 79

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20 2530

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100

105

110 <210> 80 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 80

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Ser Ala Ser Ile Ser Cys Arg Ser Ser Gln Gly Leu Leu Arg Ser 20 2530

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100

105

110 <210> 81 <211> 831 <212> PROTEIN <213> Homo sapiens <400> 81

Met Glu Arg Pro Trp Gly Ala Ala Asp Gly Leu Ser Arg Trp Pro His 1 5 1015

Gly Leu Gly Leu Leu Leu Leu Leu Gln Leu Leu Pro Pro Ser Thr Leu 20 2530

Ser Gln Asp Arg Leu Asp Ala Pro Pro Pro Pro Ala Ala Pro Leu Pro 35 4045

Arg Trp Ser Gly Pro Ile Gly Val Ser Trp Gly Leu Arg Ala Ala Ala 50 5560

Ala Gly Gly Ala Phe Pro Arg Gly Gly Arg Trp Arg Arg Ser Ala Pro 65 70 7580

- 155 046146

Gly Glu Asp Glu Glu 85

Ala Asn Asn Thr His 100

Ser Leu Ser Trp Val 115

Thr Phe His Val Pro 130

Tyr Arg Ser Glu Asp 145

Ile Asn Asn Thr Phe 165 Glu Asn Ser Gly Lys 180

Arg Gly Gly Arg Ile 195

Gln Thr Asp Leu Pro 210

Gln Asn Ser Asp Tyr 225

Val Ser Lys Asn Phe 245 Cys Leu Ala Lys Trp 260

Ala Asn Gly Ser Cys 275

Thr Ser Asp Leu Gly 290

Ser Phe Gly Leu Gly 305

Lys Asp Thr Thr Arg 325

Trp Ser Met Ala Gln 340

Ile Leu Ala Ala Asn 355

Gly Asp Thr Gly Phe 370

Val Tyr Ser Lys Ser 385

Glu Thr Asp Phe Thr 405 Ser Val Leu Ser Glu 420

Gln Gly Gly Arg Trp 435

Asp Ala Thr Ala Lys 450

Ser Tyr Ser Ile Ser 465

Glu Pro Asn Ala Val 485 Ala Ile Ser Val Met 500

Tyr Ser Trp Thr Lys 515

Asp Ser Gly Gly Ile 530

Asn Val Ile Lys Phe 545

Thr Phe Thr Arg Asp 565

Gly Ala Arg Ser Met

Cys Gly Arg Val Arg 90

Gln His Val Phe Asp 105

Gly Asp Ser Thr Gly 120

Leu Val Ile Met Thr

135

Tyr Gly Lys Asn Phe 150

Ile Arg Thr Glu Phe 170

Val Val Leu Thr Ala 185

Phe Arg Ser Ser Asp 200

Phe His Pro Leu Thr

215

Leu Leu Ala Leu Ser 230

Gly Gly Lys Trp Glu 250

Gly Ser Asp Asn Thr 265

Lys Ala Asp Leu Gly 280

Lys Ser Phe Lys Thr 295

Gly Arg Phe Leu Phe 310

Arg Ile His Val Ser 330

Leu Pro Ser Val Gly 345

Asp Asp Met Val Phe 360

Gly Thr Ile Phe Thr 375

Leu Asp Arg His Leu 390

Asn Val Thr Ser Leu 410

Asp Asn Ser Ile Gln 425

Thr His Leu Arg Lys 440

Asn Lys Asn Glu Cys 455

Gln Lys Leu Asn Val 470

Gly Ile Val Ile Ala 490

Val Pro Asp Val Tyr 505

Met Leu Glu Gly Pro 520

Ile Val Ala Ile Glu

535

Ser Thr Asp Glu Gly 550

Pro Ile Tyr Phe Thr 570

Asn Ile Ser Ile Trp

Asp Phe Val Ala Lys 95

Asp Leu Arg Gly Ser 110

Val Ile Leu Val Leu 125

Phe Gly Gln Ser Lys 140

Lys Asp Ile Thr Asp 155

Gly Met Ala Ile Gly 175

Glu Val Ser Gly Gly 190

Phe Ala Lys Asn Phe 205

Gln Met Met Tyr Ser 220

Thr Glu Asn Gly Leu 235

Glu Ile His Lys Ala 255 Ile Phe Phe Thr Thr 270

Ala Leu Glu Leu Trp 285

Ile Gly Val Lys Ile 300

Ala Ser Val Met Ala 315

Thr Asp Gln Gly Asp 335

Gln Glu Gln Phe Tyr 350

Met His Val Asp Glu 365

Ser Asp Asp Arg Gly 380

Tyr Thr Thr Thr Gly 395

Arg Gly Val Tyr Ile 415 Thr Met Ile Thr Phe 430

Pro Glu Asn Ser Glu 445

Ser Leu His Ile His 460

Pro Met Ala Pro Leu 475

His Gly Ser Val Gly 495

Ile Ser Asp Asp Gly 510

His Tyr Tyr Thr Ile 525

His Ser Ser Arg Pro 540

Gln Cys Trp Gln Thr 555

Gly Leu Ala Ser Glu 575

Gly Phe Thr Glu Ser

Leu

Val

Thr

Leu

Leu 160 Pro

Ser

Val

Pro

Trp 240 Val

Tyr

Arg

Tyr

Asp 320 Thr

Ser

Pro

Ile

Gly 400 Thr

Asp

Cys

Ala

Ser 480 Asp

Gly

Leu

Ile

Tyr 560 Pro

Phe

- 156 046146

580

Leu Thr Ser Gln 595

Glu Arg Asn Cys 610

Thr Asp Pro Glu 625

Gln Phe Leu Arg

Tyr Val Val Thr

660

Phe Leu Cys Asp 675

Val Glu Gln Pro 690

Gly Arg Glu Glu 705

Asp Lys Cys Gln

Lys Lys Lys Cys

740

Lys Ser Asn Ser 755

Val Thr Val Val 770

Gly Arg Phe Leu 785

Ala Asn Gly Val

Asn Lys Ser Gly

820

Trp

Glu

Asp

Leu 645 Lys

Phe

Glu

His

Gly 725 Thr

Val

Ala

Val

Asp 805 Tyr

585590

Val Ser Tyr Thr Ile Asp Phe Lys Asp Ile Leu 600605

Glu Lys Asp Tyr Thr Ile Trp Leu Ala His Ser 615620

Tyr Glu Asp Gly Cys Ile Leu Gly Tyr Lys Glu 630 635640

Arg Lys Ser Ser Val Cys Gln Asn Gly Arg Asp 650655

Gln Pro Ser Ile Cys Leu Cys Ser Leu Glu Asp 665670

Gly Tyr Tyr Arg Pro Glu Asn Asp Ser Lys Cys 680685

Leu Lys Gly His Asp Leu Glu Phe Cys Leu Tyr 695700

Leu Thr Thr Asn Gly Tyr Arg Lys Ile Pro Gly 710 715720

Gly Val Asn Pro Val Arg Glu Val Lys Asp Leu 730735

Ser Asn Phe Leu Ser Pro Glu Lys Gln Asn Ser 745750

Pro Ile Ile Leu Ala Ile Val Gly Leu Met Leu 760765

Gly Val Leu Ile Val Lys Lys Tyr Val Cys Gly 775780

His Arg Tyr Ser Val Leu Gln Gln His Ala Glu 790 795800

Gly Val Asp Ala Leu Asp Thr Ala Ser His Thr 810815

His Asp Asp Ser Asp Glu Asp Leu Leu Glu 825830 <210> 82 <211> 825 <212> PROTEIN <213> Mus musculus <400> 82

Met Glu Arg Pro 1

Gly Leu Leu Leu

Asp Arg Leu Asp 35

Ala Gly Pro Val 50

Gly Pro Val Pro 65

Asp Gln Asp Cys

Asn Thr His Gln

100

Ser Trp Val Gly 115

Gln Val Pro Leu 130

Ser Glu Asp Tyr 145

Asn Thr Phe Ile

Ser Gly Lys Val

180

Arg Gly Ala Ala 5

Leu Leu Gln Leu

Ala Pro Pro Pro 40

Gly Val Ser Trp 55

Arg Ala Gly Arg 70

Gly Arg Leu Pro 85

His Val Phe Asp

Asp Ser Thr Gly

120

Val Ile Val Ser 135

Gly Lys Asn Phe 150

Arg Thr Glu Phe 165

Ile Leu Thr Ala

Asp Gly Leu Leu 10

Leu Pro Pro Ala 25

Pro Ala Pro Pro

Gly Leu Arg Ala 60

Trp Arg Arg Gly 75

Asp Phe Ile Ala 90

Asp Leu Ser Gly 105

Val Ile Leu Val

Phe Gly Gln Ser

140

Lys Asp Ile Thr 155

Gly Met Ala Ile 170

Glu Val Ser Gly 185

Arg Trp Pro Leu 15

Ala Val Gly Gln 30

Leu Leu Arg Trp 45

Ala Ala Pro Gly

Ala Pro Ala Glu 80

Lys Leu Thr Asn 95

Ser Val Ser Leu

110

Leu Thr Thr Phe 125

Lys Leu Tyr Arg

Asn Leu Ile Asn

160

Gly Pro Glu Asn 175

Gly Ser Arg Gly 190

- 157 046146

Gly Arg Val Phe 195

Asp Leu Pro Phe 210

Ser Asp Tyr Leu 225

Lys Asn Phe Gly

Ala Lys Trp Gly 260

Gly Ser Cys Lys 275

Asp Leu Gly Lys 290

Gly Leu Gly Gly 305

Thr Thr Arg Arg

Met Ala Gln Leu

340

Ala Ala Asn Glu 355

Thr Gly Phe Gly 370

Ser Lys Ser Leu 385

Asp Phe Thr Asn

Leu Ser Glu Asp 420

Gly Arg Trp Glu 435

Thr Ala Lys Asn 450

Ser Ile Ser Gln 465

Asn Ala Val Gly

Ser Val Met Val 500

Trp Ala Lys Met 515

Gly Gly Ile Ile 530

Ile Lys Phe Ser 545

Thr Gln Glu Pro

Arg Ser Met Asn 580

Arg Gln Trp Val 595

Asn Cys Glu Glu 610

Pro Gly Asp Tyr 625

Leu Arg Leu Arg

Val Ala Lys Gln 660

Cys Asp Phe Gly 675

Gln Pro Glu Leu

Arg

His

Leu

Glu 245 Pro

Ala

Thr

Arg

Ile 325 Pro

Asp

Thr

Asp

Val 405 Asn

His

Lys

Lys

Ile 485 Pro

Leu

Val

Thr

Ile 565 Ile

Ser

Asp

Lys

Lys 645 Pro

Tyr

Lys

Ser Ser Asp Phe Ala Lys Asn Phe Val Gln 200205

Pro Leu Thr Gln Met Met Tyr Ser Pro Gln 215220

Ala Leu Ser Thr Glu Asn Gly Leu Trp Val 230235

Lys Trp Glu Glu Ile His Lys Ala Val Cys 250255

Asn Asn Ile Ile Phe Phe Thr Thr His Val 265270

Asp Leu Gly Ala Leu Glu Leu Trp Arg Thr 280285

Phe Lys Thr Ile Gly Val Lys Ile Tyr Ser 295300

Phe Leu Phe Ala Ser Val Met Ala Asp Lys 310315

His Val Ser Thr Asp Gln Gly Asp Thr Trp 330335

Ser Val Gly Gln Glu Gln Phe Tyr Ser Ile 345350

Met Val Phe Met His Val Asp Glu Pro Gly 360365

Ile Phe Thr Ser Asp Asp Arg Gly Ile Val 375380

Arg His Leu Tyr Thr Thr Thr Gly Gly Glu 390395

Thr Ser Leu Arg Gly Val Tyr Ile Thr Ser 410415

Ser Ile Gln Ser Met Ile Thr Phe Asp Gln 425430

Leu Arg Lys Pro Glu Asn Ser Lys Cys Asp 440445

Asn Glu Cys Ser Leu His Ile His Ala Ser 455460

Leu Asn Val Pro Met Ala Pro Leu Ser Glu 470475

Val Ile Ala His Gly Ser Val Gly Asp Ala 490495

Asp Val Tyr Ile Ser Asp Asp Gly Gly Tyr 505510

Glu Gly Pro His Tyr Tyr Thr Ile Leu Asp 520525

Ala Ile Glu His Ser Asn Arg Pro Ile Asn 535540

Asp Glu Gly Gln Cys Trp Gln Ser Tyr Val 550555

Tyr Phe Thr Gly Leu Ala Ser Glu Pro Gly 570575

Ser Ile Trp Gly Phe Thr Glu Ser Phe Ile 585590

Tyr Thr Val Asp Phe Lys Asp Ile Leu Glu 600605

Asp Tyr Thr Thr Trp Leu Ala His Ser Thr 615620

Asp Gly Cys Ile Leu Gly Tyr Lys Glu Gln 630635

Ser Ser Val Cys Gln Asn Gly Arg Asp Tyr 650655

Ser Val Cys Pro Cys Ser Leu Glu Asp Phe 665670

Phe Arg Pro Glu Asn Ala Ser Glu Cys Val 680685

Gly His Glu Leu Glu Phe Cys Leu Tyr Gly

Thr

Asn

Ser 240 Leu

Asn

Ser

Phe

Asp 320 Ser

Leu

Asp

Tyr

Thr 400 Thr

Gly

Ala

Tyr

Pro 480

Ile

Ser

Ser

Val

Phe 560 Ala

Thr

Arg

Asp

Phe 640 Val

Leu

Glu

Lys

- 158 046146

690 695

Glu Glu His Leu Thr Thr Asn 705 710

Cys Gln Gly Gly Met Asn Pro 725

Lys Cys Thr Ser Asn Phe Leu 740

Asn Ser Val Pro Ile Ile Leu 755

Val Val Ala Gly Val Leu Ile 770 775

Phe Leu Val His Arg Tyr Ser 785 790

Gly Val Glu Ala Leu Asp Ser 805

Asp Asp Ser Asp Glu Asp Leu 820

700

Gly Tyr Arg Lys Ile Pro Gly Asp Lys 715720

Ala Arg Glu Val Lys Asp Leu Lys Lys 730735

Asn Pro Thr Lys Gln Asn Ser Lys Ser 745750

Ala Ile Val Gly Leu Met Leu Val Thr 760765

Val Lys Lys Tyr Val Cys Gly Gly Arg 780

Val Leu Gln Gln His Ala Glu Ala Asp 795800

Thr Ser His Ala Lys Ser Gly Tyr His 810815

Leu Glu

825 <210> 83 <211> 825 <212> PROTEIN <213> Rattus norvegicus <400> 83

Met Glu Arg Pro Arg Gly Ala Ala 15

Gly Leu Leu Leu Leu Leu Gln Leu 20

Asp Arg Leu Asp Ala Pro Pro Pro 3540

Ala Gly Pro Val Gly Val Ser Trp 5055

Gly Pro Val Pro Arg Ala Gly Arg 6570

Asp Gln Asp Cys Gly Arg Leu Pro 85

Asn Thr His Gln His Val Phe Asp 100

Ser Trp Val Gly Asp Ser Thr Gly 115120

Gln Val Pro Leu Val Ile Val Ser 130135

Ser Glu Asp Tyr Gly Lys Asn Phe 145150

Asn Thr Phe Ile Arg Thr Glu Phe 165

Ser Gly Lys Val Ile Leu Thr Ala 180

Gly Arg Val Phe Arg Ser Ser Asp 195200

Asp Leu Pro Phe His Pro Leu Thr 210215

Ser Asp Tyr Leu Leu Ala Leu Ser 225230

Lys Asn Phe Gly Glu Lys Trp Glu 245

Ala Lys Trp Gly Pro Asn Asn Ile 260

Gly Ser Cys Lys Ala Asp Leu Gly 275280

Asp Leu Gly Lys Thr Phe Lys Thr 290295

Asp Gly Leu Leu Arg Trp Pro Leu 1015

Leu Pro Pro Ala Ala Val Gly Gln 2530

Pro Ala Pro Pro Leu Leu Arg Trp 45

Gly Leu Arg Ala Ala Ala Pro Gly 60

Trp Arg Arg Gly Ala Pro Ala Glu 7580

Asp Phe Ile Ala Lys Leu Thr Asn 9095

Asp Leu Ser Gly Ser Val Ser Leu 105110

Val Ile Leu Val Leu Thr Thr Phe 125

Phe Gly Gln Ser Lys Leu Tyr Arg 140

Lys Asp Ile Thr Asn Leu Ile Asn 155160

Gly Met Ala Ile Gly Pro Glu Asn 170175

Glu Val Ser Gly Gly Ser Arg Gly 185190

Phe Ala Lys Asn Phe Val Gln Thr 205

Gln Met Met Tyr Ser Pro Gln Asn 220

Thr Glu Asn Gly Leu Trp Val Ser 235240

Glu Ile His Lys Ala Val Cys Leu 250255

Ile Phe Phe Thr Thr His Val Asn

265270

Ala Leu Glu Leu Trp Arg Thr Ser 285

Ile Gly Val Lys Ile Tyr Ser Phe 300

- 159 046146

Gly Leu Gly Gly Arg 305

Thr Thr Arg Arg Ile 325

Met Ala Gln Leu Pro

340

Ala Ala Asn Asp Asp 355

Thr Gly Phe Gly Thr 370

Ser Lys Ser Leu Asp 385

Asp Phe Thr Asn Val 405

Leu Ser Glu Asp Asn 420

Gly Arg Trp Glu His 435

Thr Ala Lys Asn Lys 450

Ser Ile Ser Gln Lys 465

Asn Ala Val Gly Ile 485

Ser Val Met Val Pro 500

Trp Ala Lys Met Leu 515

Gly Gly Ile Ile Val 530

Ile Lys Phe Ser Thr 545

Ser Gln Glu Pro Val

565

Arg Ser Met Asn Ile 580

Arg Gln Trp Val Ser 595

Asn Cys Glu Glu Asn 610

Pro Gly Asp Tyr Lys 625

Leu Arg Leu Arg Lys 645

Val Ala Lys Gln Pro 660

Cys Asp Phe Gly Tyr 675

Gln Pro Glu Leu Lys 690

Glu Glu His Leu Thr 705

Cys Gln Gly Gly Met 725

Lys Cys Thr Ser Asn 740

Ser Ser Val Pro Ile 755

Val Val Ala Gly Val 770

Phe Leu Val His Arg 785

Gly Val Glu Ala Leu

Phe Leu Phe Ala Ser 310

His Val Ser Thr Asp

330

Ser Val Gly Gln Glu 345

Met Val Phe Met His 360

Ile Phe Thr Ser Asp 375

Arg His Leu Tyr Thr 390

Thr Ser Leu Arg Gly 410

Ser Ile Gln Ser Met 425

Leu Gln Lys Pro Glu 440

Asn Glu Cys Ser Leu 455

Leu Asn Val Pro Met 470

Val Ile Ala His Gly

490

Asp Val Tyr Ile Ser 505

Glu Gly Pro His Tyr 520

Ala Ile Glu His Ser 535

Asp Glu Gly Gln Cys 550

Tyr Phe Thr Gly Leu 570

Ser Ile Trp Gly Phe 585

Tyr Thr Ile Asp Phe 600

Asp Tyr Thr Thr Trp 615

Asp Gly Cys Ile Leu 630

Ser Ser Val Cys Gln 650

Ser Ile Cys Pro Cys 665

Phe Arg Pro Glu Asn 680

Gly His Glu Leu Glu 695

Thr Asn Gly Tyr Arg 710

Asn Pro Ala Arg Glu 730

Phe Leu Asn Pro Lys 745

Ile Leu Ala Ile Val 760

Leu Ile Val Lys Lys 775

Tyr Ser Val Leu Gln 790

Asp Thr Ala Ser His

Val Met Ala Asp Lys 315

Gln Gly Asp Thr Trp 335

Gln Phe Tyr Ser Ile 350

Val Asp Glu Pro Gly 365

Asp Arg Gly Ile Val 380

Thr Thr Gly Gly Glu 395

Val Tyr Ile Thr Ser 415

Ile Thr Phe Asp Gln 430

Asn Ser Lys Cys Asp 445

His Ile His Ala Ser 460

Ala Pro Leu Ser Glu 475

Ser Val Gly Asp Ala 495

Asp Asp Gly Gly Tyr 510

Tyr Thr Ile Leu Asp 525

Asn Arg Pro Ile Asn 540

Trp Gln Ser Tyr Val 555

Ala Ser Glu Pro Gly 575

Thr Glu Ser Phe Leu

590

Lys Asp Ile Leu Glu 605

Leu Ala His Ser Thr 620

Gly Tyr Lys Glu Gln 635

Asn Gly Arg Asp Tyr 655

Ser Leu Glu Asp Phe 670

Ala Ser Glu Cys Val 685

Phe Cys Leu Tyr Gly 700

Lys Ile Pro Gly Asp 715

Val Lys Asp Leu Lys 735

Lys Gln Asn Ser Lys 750

Gly Leu Met Leu Val 765

Tyr Val Cys Gly Gly 780

Gln His Ala Glu Ala 795

Ala Lys Ser Gly Tyr

Asp 320 Ser

Leu

Asp

Tyr

Thr 400 Thr

Gly

Ala

Tyr

Pro 480 Ile

Ser

Ser

Val

Phe 560 Ala

Thr

Arg

Asp

Phe 640 Val

Leu

Glu

Lys

Arg 720 Lys

Ser

Thr

Arg

Asp 800 His

- 160 046146

805 810

Asp Asp Ser Asp Glu Asp Leu Leu Glu 820 825

815 <210> 84 <211> 8 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

< 221> VARIANT < 222> 1 < 223> Xaa = Ser or Thr <220>

< 221> VARIANT < 222> 2 < 223> Xaa = any amino acid <220>

< 221> VARIANT < 222> 3 < 223> Xaa = Asp or Asn <220>

< 221> VARIANT < 222> 4, 5, 6, 7 < 223> Xaa = any amino acid <220>

< 221> VARIANT < 222> 8 < 223> Xaa = Trp, Phe, or Tyr < 400> 84

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

5 <210> 85 <211> 11 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <220>

<221> VARIANT <222> 1, 2 <223> Xaa = any amino acid <220>

<221> VARIANT <222> 3 <223> Xaa = Ser or Thr <220>

<221> VARIANT

- 161 046146 <222> 4 <223> Xaa = any amino acid <220>

<221> VARIANT <222> 5 <223> Xaa = Asp or Asn <220>

<221> VARIANT <222> 6, 8 <223> Xaa = any amino acid <220>

<221> VARIANT <222> 9 <223> Xaa = Thr or Ser <220>

<221> VARIANT <222> 10 <223> Xaa = Trp, Phe, or Tyr <220>

<221> VARIANT <222> 11 <223> Xaa = any amino acid <400> 85

Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa

5 10 <210> 86 <211> 8 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 86

Ser Ser Asp Phe Ala Lys Asn Phe

5 <210> 87 <211> 6 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 87

Asn Gln Phe Ser Leu Lys

5 <210> 88 <211> 23 <212> PROTEIN <213> Artificial sequence

- 162 046146 <220>

<223> Synthetic construct <400> 88

Gln Gly Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr 1 5 10 15

Val Ser Ser Ala Ser Thr Lys <210> 89 <211> 49 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 89

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu 1 5 10 15 Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu 20 25 thirty Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr 35 40 45

Lys <210> 90 <211> 7 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 90

Asp Thr Leu Met Ile Ser Arg

5 <210> 91 <211> 33 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 91

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro 1 5 10 15 Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 20 25 thirty

Lys <210> 92 <211> 16 <212> PROTEIN

- 163 046146 <213> Artificial sequence <220>

<223> Synthetic construct <400> 92

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

5 10 15 <210> 93 <211> 16 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 93

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr 1 5 10 15 <210> 94 <211> 10 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 94

Asn Gln Val Ser Leu Thr Cys Leu Val Lys

5 10 <210> 95 <211> 22 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 95

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 1 5 10 15

Pro Glu Asn Asn Tyr Lys 20 <210> 96 <211> 23 <212> PROTEIN <213> Artificial Sequence <220>

<223> Synthetic construct <400> 96

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 1 5 10 15

His Asn His Tyr Thr Gln Lys

Lys

Lys

Gln

Leu

- 164 046146 <210> 97 <211> 24 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 97

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg <210> 98 <211> 7 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 98

Ser Ser Gln Ser Leu Leu Arg

5 <210> 99 <211> 28 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 99

Ser Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln 1 5 10 15

Ser Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg 20 25 <210> 100 <211> 26 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 100

Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gln Gln Glu 1 5 10 15

Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys

25 <210> 101 <211> 16

- 165 046146 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 101

Ser Gly Thr Ala Ser

5 sequence construct

Val Val Cys Leu Leu

Asn Asn Phe Tyr Pro

Arg <210> 102 <211> 20 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 102

Val Asp Asn Ala Leu 1 5

Gln Asp Ser Lys sequence construct

Gln Ser Gly Asn Ser

Gln Glu Ser Val Thr

Glu <210> 103 <211> 330 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 103

Ala Ser Thr Lys Gly 1 5

Ser Thr Ser Gly Gly 20

Phe Pro Glu Pro Val 35

Gly Val His Thr Phe 50

Leu Ser Ser Val Val 65

Tyr Ile Cys Asn Val 85

Lys Val Glu Pro Lys 100

Pro Ala Pro Glu Ala

115

Lys Pro Lys Asp Thr 130

Val Val Val Asp Val 145

Tyr Val Asp Gly Val 165

Glu Gln Tyr Asn Ser 180

His Gln Asp Trp Leu 195

Lys Ala Leu Pro Ala 210 sequence construct

Pro Ser Val Phe Pro 10

Thr Ala Ala Leu Gly 25

Thr Val Ser Trp Asn 40

Pro Ala Val Leu Gln 55

Thr Val Pro Ser Ser 70

Asn His Lys Pro Ser 90

Ser Cys Asp Lys Thr 105

Ala Gly Gly Pro Ser 120

Leu Met Ile Ser Arg 135

Ser His Glu Asp Pro 150

Glu Val His Asn Ala 170

Thr Tyr Arg Val Val 185

Asn Gly Lys Glu Tyr 200

Ser Ile Glu Lys Thr 215

Leu Ala Pro Ser Ser 15

Cys Leu Val Lys Asp 30

Ser Gly Ala Leu Thr 45

Ser Ser Gly Leu Tyr 60

Ser Leu Gly Thr Gln 75

Asn Thr Lys Val Asp 95

His Thr Cys Pro Pro 110

Val Phe Leu Phe Pro 125

Thr Pro Glu Val Thr 140

Glu Val Lys Phe Asn 155

Lys Thr Lys Pro Arg 175

Ser Val Leu Thr Val 190

Lys Cys Lys Val Ser 205

Ile Ser Lys Ala Lys 220

Lys

Tyr

Ser

Ser

Thr 80 Lys

Cys

Pro

Cys

Trp 160 Glu

Leu

Asn

Gly

- 166 046146

Gln 225 Leu

Pro

Asn

Leu

Val 305 Gln

Pro Arg Glu Pro Gln 230

Thr Lys Asn Gln Val 245

Ser Asp Ile Ala Val 260

Tyr Lys Thr Thr Pro 275

Tyr Ser Lys Leu Thr 290

Phe Ser Cys Ser Val 310

Lys Ser Leu Ser Leu 325

Val Tyr Thr Leu

Ser Leu Thr Cys 250

Glu Trp Glu Ser 265

Pro Val Leu Asp 280

Val Asp Lys Ser 295

Met His Glu Ala

Ser Pro Gly Lys 330

Pro Pro Ser Arg Asp 235

Leu Val Lys Gly Phe 255

Asn Gly Gln Pro Glu 270

Ser Asp Gly Ser Phe 285

Arg Trp Gln Gln Gly 300

Leu His Asn His Tyr 315

Glu 240 Tyr

Asn

Phe

Asn

Thr 320 <210> 104 <211> 329 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 104

Ala Ser Thr Lys Gly 1 5

Ser Thr Ser Gly Gly 20

Phe Pro Glu Pro Val 35

Gly Val His Thr Phe 50

Leu Ser Ser Val Val 65

Tyr Ile Cys Asn Val 85

Lys Val Glu Pro Lys 100

Pro Ala Pro Glu Ala

115

Lys Pro Lys Asp Thr 130

Val Val Val Asp Val 145

Tyr Val Asp Gly Val 165

Glu Gln Tyr Asn Ser 180

His Gln Asp Trp Leu 195

Lys Ala Leu Pro Ala 210

Gln Pro Arg Glu Pro 225

Leu Thr Lys Asn Gln 245

Pro Ser Asp Ile Ala 260

Asn Tyr Lys Thr Thr 275 Leu Tyr Ser Lys Leu 290 sequence construct

Pro Ser Val Phe Pro 10

Thr Ala Ala Leu Gly 25

Thr Val Ser Trp Asn 40

Pro Ala Val Leu Gln 55

Thr Val Pro Ser Ser 70

Asn His Lys Pro Ser 90

Ser Cys Asp Lys Thr 105

Ala Gly Gly Pro Ser 120

Leu Met Ile Ser Arg 135

Ser His Glu Asp Pro 150

Glu Val His Asn Ala 170

Thr Tyr Arg Val Val 185

Asn Gly Lys Glu Tyr 200

Ser Ile Glu Lys Thr 215

Gln Val Tyr Thr Leu 230

Val Ser Leu Thr Cys 250

Val Glu Trp Glu Ser 265

Pro Pro Val Leu Asp 280

Thr Val Asp Lys Ser 295

Leu Ala Pro Ser Ser 15

Cys Leu Val Lys Asp 30

Ser Gly Ala Leu Thr 45

Ser Ser Gly Leu Tyr 60

Ser Leu Gly Thr Gln 75

Asn Thr Lys Val Asp 95

His Thr Cys Pro Pro 110

Val Phe Leu Phe Pro 125

Thr Pro Glu Val Thr

140

Glu Val Lys Phe Asn 155

Lys Thr Lys Pro Arg 175

Ser Val Leu Thr Val 190

Lys Cys Lys Val Ser 205

Ile Ser Lys Ala Lys 220

Pro Pro Ser Arg Asp 235

Leu Val Lys Gly Phe 255

Asn Gly Gln Pro Glu 270

Ser Asp Gly Ser Phe 285

Arg Trp Gln Gln Gly 300

Lys

Tyr

Ser

Ser

Thr 80 Lys

Cys

Pro

Cys

Trp 160 Glu

Leu

Asn

Gly

Glu 240 Tyr

Asn

Phe

Asn

- 167 046146

Val Phe Ser Cys 305

Gln Lys Ser Leu

Ser Val Met His Glu Ala

310

Ser Leu Ser Pro Gly

325

Leu His Asn His Tyr 315

Thr

320 <210> 105 <211> 452 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 105

Gln Val Gln Leu Gln 1 5

Thr Leu Ser Leu Thr 20

Tyr Tyr Trp Gly Trp 35

Ile Gly Thr Ile Tyr 50

Lys Ser Gln Val Thr 65

Leu Glu Leu Ser Ser 85

Ala Arg Gln Gly Ser 100

Gly Gln Gly Thr Thr 115 Ser Val Phe Pro Leu 130

Ala Ala Leu Gly Cys 145

Val Ser Trp Asn Ser 165

Ala Val Leu Gln Ser 180

Val Pro Ser Ser Ser 195

His Lys Pro Ser Asn 210

Cys Asp Lys Thr His 225

Gly Gly Pro Ser Val 245

Met Ile Ser Arg Thr 260

His Glu Asp Pro Glu 275

Val His Asn Ala Lys 290

Tyr Arg Val Val Ser 305

Gly Lys Glu Tyr Lys 325

Ile Glu Lys Thr Ile 340

Val Tyr Thr Leu Pro 355

Ser Leu Thr Cys Leu 370 sequence construct

Glu Ser Gly Pro Gly 10

Cys Ala Val Ser Gly 25

Ile Arg Gln Pro Pro 40

His Ser Gly Ser Thr 55

Ile Ser Val Asp Thr 70

Val Thr Ala Ala Asp 90

Ile Gln Gln Gly Tyr 105

Val Thr Val Ser Ser 120

Ala Pro Ser Ser Lys 135

Leu Val Lys Asp Tyr 150

Gly Ala Leu Thr Ser 170

Ser Gly Leu Tyr Ser 185

Leu Gly Thr Gln Thr 200

Thr Lys Val Asp Lys 215

Thr Cys Pro Pro Cys 230

Phe Leu Phe Pro Pro 250

Pro Glu Val Thr Cys 265

Val Lys Phe Asn Trp 280

Thr Lys Pro Arg Glu 295

Val Leu Thr Val Leu 310

Cys Lys Val Ser Asn 330

Ser Lys Ala Lys Gly 345

Pro Ser Arg Asp Glu 360

Val Lys Gly Phe Tyr 375

Leu Val Lys Pro Ser 15

Tyr Ser Ile Ser Ser 30

Gly Lys Gly Leu Glu 45

Tyr Tyr Asn Pro Ser 60

Ser Lys Asn Gln Phe 75

Thr Ala Val Tyr Tyr 95

Tyr Gly Met Asp Val 110

Ala Ser Thr Lys Gly 125

Ser Thr Ser Gly Gly 140

Phe Pro Glu Pro Val 155

Gly Val His Thr Phe 175

Leu Ser Ser Val Val 190

Tyr Ile Cys Asn Val 205

Lys Val Glu Pro Lys 220

Pro Ala Pro Glu Ala 235

Lys Pro Lys Asp Thr 255

Val Val Val Asp Val 270

Tyr Val Asp Gly Val 285

Glu Gln Tyr Asn Ser 300

His Gln Asp Trp Leu 315

Lys Ala Leu Pro Ala 335

Gln Pro Arg Glu Pro 350

Leu Thr Lys Asn Gln 365

Pro Ser Asp Ile Ala 380

Glu

Gly

Trp

Leu

Ser 80 Cys

Trp

Pro

Thr

Thr 160 Pro

Thr

Asn

Ser

Ala 240 Leu

Ser

Glu

Thr

Asn 320 Ser

Gln

Val

Val

- 168 046146

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390395

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

420 425430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440445

Ser Pro Gly Lys

450

Pro 400 Thr

Val

Leu <210> 106 <211> 451 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 106

Gln Val Gln Leu Gln 1 5

Thr Leu Ser Leu Thr

Tyr Tyr Trp Gly Trp 35

Ile Gly Thr Ile Tyr 50

Lys Ser Gln Val Thr 65

Leu Glu Leu Ser Ser 85

Ala Arg Gln Gly Ser 100

Gly Gln Gly Thr Thr 115

Ser Val Phe Pro Leu

130

Ala Ala Leu Gly Cys 145

Val Ser Trp Asn Ser 165

Ala Val Leu Gln Ser 180

Val Pro Ser Ser Ser 195

His Lys Pro Ser Asn 210

Cys Asp Lys Thr His 225

Gly Gly Pro Ser Val 245

Met Ile Ser Arg Thr 260

His Glu Asp Pro Glu 275

Val His Asn Ala Lys 290

Tyr Arg Val Val Ser 305

Gly Lys Glu Tyr Lys

325 sequence construct

Glu Ser Gly Pro Gly 10

Cys Ala Val Ser Gly 25

Ile Arg Gln Pro Pro 40

His Ser Gly Ser Thr 55

Ile Ser Val Asp Thr 70

Val Thr Ala Ala Asp 90

Ile Gln Gln Gly Tyr 105

Val Thr Val Ser Ser 120

Ala Pro Ser Ser Lys 135

Leu Val Lys Asp Tyr 150

Gly Ala Leu Thr Ser 170

Ser Gly Leu Tyr Ser 185

Leu Gly Thr Gln Thr 200

Thr Lys Val Asp Lys 215

Thr Cys Pro Pro Cys 230

Phe Leu Phe Pro Pro 250

Pro Glu Val Thr Cys 265

Val Lys Phe Asn Trp 280

Thr Lys Pro Arg Glu 295

Val Leu Thr Val Leu 310

Cys Lys Val Ser Asn 330

Leu Val Lys Pro Ser 15

Tyr Ser Ile Ser Ser 30

Gly Lys Gly Leu Glu 45

Tyr Tyr Asn Pro Ser 60

Ser Lys Asn Gln Phe 75

Thr Ala Val Tyr Tyr 95

Tyr Gly Met Asp Val 110

Ala Ser Thr Lys Gly 125

Ser Thr Ser Gly Gly 140

Phe Pro Glu Pro Val 155

Gly Val His Thr Phe 175

Leu Ser Ser Val Val 190

Tyr Ile Cys Asn Val 205

Lys Val Glu Pro Lys 220

Pro Ala Pro Glu Ala 235

Lys Pro Lys Asp Thr 255

Val Val Val Asp Val 270

Tyr Val Asp Gly Val 285

Glu Gln Tyr Asn Ser 300

His Gln Asp Trp Leu 315

Lys Ala Leu Pro Ala 335

Glu

Gly

Trp

Leu

Ser 80

Cys

Trp

Pro

Thr

Thr 160 Pro

Thr

Asn

Ser

Ala 240 Leu

Ser

Glu

Thr

Asn 320 Ser

- 169 046146

Ile Glu Lys Thr Ile 340

Val Tyr Thr Leu Pro 355

Ser Leu Thr Cys Leu 370

Glu Trp Glu Ser Asn 385

Pro Val Leu Asp Ser 405

Val Asp Lys Ser Arg 420

Met His Glu Ala Leu 435

Ser Pro Gly 450

Ser Lys Ala Lys Gly 345

Pro Ser Arg Asp Glu 360

Val Lys Gly Phe Tyr 375

Gly Gln Pro Glu Asn 390

Asp Gly Ser Phe Phe 410

Trp Gln Gln Gly Asn 425

His Asn His Tyr Thr 440

Gln Pro Arg Glu Pro 350

Leu Thr Lys Asn Gln 365

Pro Ser Asp Ile Ala 380

Asn Tyr Lys Thr Thr 395

Leu Tyr Ser Lys Leu 415

Val Phe Ser Cys Ser 430

Gln Lys Ser Leu Ser 445

Gln

Val

Val

Pro 400 Thr

Val

Leu <210> 107 <211> 122 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 107

Gln Val Gln Leu Gln 1 5

Thr Leu Ser Leu Thr

Arg Tyr Trp Gly Trp 35

Ile Gly Ser Ile Tyr 50

Lys Ser Arg Val Thr 65

Leu Lys Leu Ser Ser 85

Ala Arg Gln Gly Ser 100

Gly Gln Gly Thr Thr 115 sequence construct

Glu Ser Gly Pro Gly 10

Cys Ala Val Ser Gly 25

Ile Arg Gln Pro Pro 40

His Ser Gly Ser Thr 55

Ile Ser Val Asp Thr 70

Val Thr Ala Ala Asp 90

Ile Lys Gln Gly Tyr 105

Val Thr Val Ser Ser 120

Leu Val Lys Pro Ser 15

Tyr Ser Ile Ser Ser 30

Gly Lys Gly Leu Glu 45

Tyr Tyr Asn Pro Ser 60

Ser Lys Asn Gln Phe 75

Thr Ala Val Tyr Tyr 95

Tyr Gly Met Asp Val 110

Glu

Val

Trp

Leu

Ser 80 Cys

Trp <210> 108 <211> 122 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 108

Gln Val Gln Leu Gln 1 5

Thr Leu Ser Leu Thr

Arg Tyr Trp Gly Trp 35

Ile Gly Ala Ile Tyr

Lys Ser Arg Val Thr sequence construct

Glu Ser Gly Pro Gly 10

Cys Ala Val Ser Gly 25

Ile Arg Gln Pro Pro 40

Pro Ser Gly Ser Thr 55

Ile Ser Val Asp Thr

Leu Val Lys Pro Ser 15

Tyr Ser Ile Ser Ser 30

Gly Lys Gly Leu Glu 45

Tyr Tyr Asn Pro Ser 60

Ser Lys Asn Gln Phe

Glu

Val

Trp

Leu

Ser

- 170 046146

70

Leu Lys Leu Ser Ser Val 85

Ala Arg Gln Gly Ser Ile 100

Gly Gln Gly Thr Thr Val 115

7580

Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Lys Gln Gly Tyr Tyr Gly Met Asp Val Trp 105110

Thr Val Ser Ser

120 <210> 109 <211> 122 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 109

Gln Val Gln Leu Gln Glu Ser Gly 15

Thr Leu Ser Leu Thr Cys Ala Val 20

Tyr Tyr Trp Gly Trp Ile Arg Gln 3540

Ile Gly Thr Ile Tyr His Ser Gly 5055

Lys Ser Arg Val Thr Ile Ser Val 6570

Leu Lys Leu Ser Ser Val Thr Ala 85

Ala Arg Gln Gly Ser Ile Lys Gln 100

Gly Gln Gly Thr Thr Val Thr Val

115120

Pro Gly Leu Val Lys Pro Ser Glu 1015

Ser Gly Tyr Ser Ile Ser Ser Gly 2530

Pro Pro Gly Lys Gly Leu Glu Trp 45

Ser Thr Tyr Tyr Asn Pro Ser Leu 60

Asp Thr Ser Lys Asn Gln Phe Ser 7580

Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Gly Tyr Tyr Gly Met Asp Val Trp 105110

Ser Ser <210> 110 <211> 122 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 110

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 1015

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 2530

Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 4045

Ile Gly Thr Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 5560

Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln PheSer

70 7580

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr TyrCys

9095

Ala Arg Gln Gly Ser Ile Lys Gln Gly Tyr Tyr Gly Met Asp Val Trp 100 105110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115120

- 171 046146 <210> 111 <211> 122 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 111

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 1015

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 20 2530

Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 4045

Ile Gly Thr Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 50 5560

Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln PheSer

70 7580

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr TyrCys

9095

Ala Arg Gln Gly Ser Ile Lys Gln Gly Tyr Tyr Gly Met Asp Val Trp 100 105110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115120 <210> 112 <211> 122 <212> PROTEIN <213>

Artificial sequence <220>

<223> Synthetic construct <400> 112

Gln Val Gln Leu Gln Glu Ser Gly 15

Thr Leu Ser Leu Thr Cys Ala Val 20

Tyr Tyr Trp Gly Trp Ile Arg Gln 3540

Ile Gly Thr Ile Tyr His Ser Gly 5055

Lys Ser Arg Val Thr Ile Ser Val

6570

Leu Lys Leu Ser Ser Val Thr Ala 85

Ala Arg Gln Gly Ser Ile Lys Gln 100

Gly Gln Gly Thr Thr Val Thr Val

115120

Pro Gly Leu Val Lys Pro Ser Glu 1015

Ser Gly Tyr Ser Ile Ser Ser Gly 2530

Pro Pro Gly Lys Gly Leu Glu Trp 45

Ser Thr Tyr Tyr Asn Pro Ser Leu 60

Asp Thr Ser Lys Asn Gln Phe Ser 7580

Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Gly Tyr Tyr Gly Met Asp Val Trp 105110

Ser Ser <210> 113 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 113

- 172 046146

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Thr Pro Leu 100

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu His 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln 4045

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys 7075

Glu Asp Val Gly Val Tyr Tyr Cys Met Gln 9095

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 105110

Gly

Ser

Ser

Pro

Ile 80

Gln

Lys <210> 114 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223>

<400> 114

Asp Ile Val 1

Glu Pro Ala

Asn Gly Tyr 35

Pro Gln Leu 50

Asp Arg Phe 65

Ser Arg Val

Gln Glu Thr

Synthetic construct

Met Thr Gln 5

Ser Ile Ser 20

Asn Tyr Leu

Leu Ile Tyr

Ser Gly Ser 70

Glu Ala Glu 85

Pro Leu Thr 100

Ser Pro Leu

Cys Arg Ser 25

Asp Trp Tyr 40

Leu Gly Ser 55

Gly Ser Gly

Asp Val Gly

Phe Gly Gly

105

Ser Leu Pro 10

Ser Gln Ser

Leu Gln Lys

Asn Arg Ala 60

Thr Asp Phe 75

Val Tyr Tyr 90

Gly Thr Lys

Val Thr Pro 15

Leu Leu His 30

Pro Gly Gln 45

Ser Gly Val

Thr Leu Lys

Cys Met Gln 95

Val Glu Ile

110

Gly

Ser

Ser

Pro

Ile 80

Gln

Lys <210> 115 <211> 112 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 115

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Thr Pro Leu sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Val Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile

Gly

Ser

Pro

Pro

Ile 80

Gln

Lys

- 173 046146

100 105 110

116

112

PROTEIN

Artificial <210>

<211>

<212>

<213>

<220>

<223>

<400> 116

Asp Ile Val Met 1

Glu Ser Ala Ser

Asn Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Val Glu

Gln Glu Thr Pro

100

Synthetic sequence construct

Pro

Arg

Thr Gln Ser 5

Ile Ser Cys

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Leu Ser Leu Pro 10

Ser Ser Gln Gly 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110

117

112

PROTEIN

Artificial <210>

<211>

<212>

<213>

<220>

<223>

<400> 117

Asp Ile Val Met 1

Glu Pro Ala Ser

Asn Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Val Glu

Gln Glu Thr Pro

100

Synthetic sequence construct

Pro

Arg

Thr Gln Ser 5

Ile Ser Cys

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110 <210> 118 <211> 112 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 118

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

- 174 046146

Gly Pro Ala Ser 20

Asn Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Val Glu

Gln Glu Thr Pro 100

Ile Ser Cys Arg

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

15

Ser Ser Gln Ser Leu Leu Arg 25 30

Tyr Leu Gln Lys Pro Gly Gln 45

Ser Asn Arg Ala Ser Gly Val 60

Gly Thr Asp Phe Thr Leu Lys 75

Gly Val Tyr Tyr Cys Met Gln 90 95

Gly Gly Thr Lys Val Glu Ile 105 110

Ser

Ser

Pro

Ile 80 Gln

Lys <210> 119 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 119

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

His Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln

165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val

135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu

170

Ser Thr Leu Thr Leu

185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 120 <211> 219 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 120

- 175 046146

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Lys Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 115 120125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 130 135140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala LeuGln

145 150 155160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys AspSer

165 170175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210

215 <210> 121 <211> 219 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 121

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Gln Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Ala Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met GlnGln

9095

Gln Glu Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 115 120125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 130 135140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala LeuGln

145 150 155160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys AspSer

165 170175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180

185

190

- 176 046146

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

200205

Phe Asn Arg Gly Glu Cys

215 <210> 122 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 122

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile

Tyr Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 4045

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 70 7580

Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gln 9095

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 105110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 120125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 135140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 150 155160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 170175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 185190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 200205

Phe Asn Arg Gly Glu Cys 215 <210> 123 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 123

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile

Glu Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly sequence construct

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 4045

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

- 177 046146

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu

170

Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 124 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 124

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Trp Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu 170 Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 125 <211> 219 <212> PROTEIN

- 178 046146 <213> Artificial sequentially <220>

<223> Synthetic construct <400> 125

Asp Ile Val Met Thr Gln Ser Pro Leu 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser 2025

Phe Gly Tyr Asn Tyr Leu Asp Trp Tyr 3540

Pro Gln Leu Leu Ile Tyr Leu Gly Ser 5055

Asp Arg Phe Ser Gly Ser Gly Ser Gly 6570

Ser Arg Ala Glu Ala Glu Asp Val Gly 85

Gln Glu Ala Pro Leu Thr Phe Gly Gly 100105

Arg Thr Val Ala Ala Pro Ser Val Phe 115120

Gln Leu Lys Ser Gly Thr Ala Ser Val 130135

Tyr Pro Arg Glu Ala Lys Val Gln Trp

145150

Ser Gly Asn Ser Gln Glu Ser Val Thr 165

Thr Tyr Ser Leu Ser Ser Thr Leu Thr 180185

Lys His Lys Val Tyr Ala Cys Glu Val 195200

Pro Val Thr Lys Ser Phe Asn Arg Gly 210215 <210> 126 <211> 219 <212> PROTEIN <213> Artificial sequentially <220>

<223> Synthetic construct <400> 126

Asp Ile Val Met Thr Gln Ser Pro Leu 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser 2025

Ile Gly Tyr Asn Tyr Leu Asp Trp Tyr 3540

Pro Gln Leu Leu Ile Tyr Leu Gly Ser 5055

Asp Arg Phe Ser Gly Ser Gly Ser Gly 6570

Ser Arg Ala Glu Ala Glu Asp Val Gly 85

Gln Glu Ala Pro Leu Thr Phe Gly Gly 100105

Arg Thr Val Ala Ala Pro Ser Val Phe 115120

Gln Leu Lys Ser Gly Thr Ala Ser Val 130135

Leu Pro Val Thr Pro Gly 15

Gln Ser Leu Leu Arg Ser 30

Gln Lys Pro Gly Gln Ser 45

Arg Ala Ser Gly Val Pro 60

Asp Phe Thr Leu Lys Ile 75 80

Tyr Tyr Cys Met Gln Gln 95

Thr Lys Val Glu Ile Lys 110

Phe Pro Pro Ser Asp Glu 125

Cys Leu Leu Asn Asn Phe 140

Val Asp Asn Ala Leu Gln 155 160

Gln Asp Ser Lys Asp Ser 175

Ser Lys Ala Asp Tyr Glu 190

His Gln Gly Leu Ser Ser 205

Cys

Leu Pro Val Thr Pro Gly 15

Gln Ser Leu Leu Arg Ser 30

Gln Lys Pro Gly Gln Ser 45

Arg Ala Ser Gly Val Pro 60

Asp Phe Thr Leu Lys Ile 75 80

Tyr Tyr Cys Met Gln Gln 95

Thr Lys Val Glu Ile Lys 110

Phe Pro Pro Ser Asp Glu 125

Cys Leu Leu Asn Asn Phe 140

- 179 046146

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys AspSer

165 170175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210215 <210> 127 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 127

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Val Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser Leu 10

Ser Cys Arg Ser Ser Gln 25

Leu Asp Trp Tyr Leu Gln 40

Tyr Leu Gly Ser Asn Arg 55

Ser Gly Ser Gly Thr Asp 70 75

Glu Asp Val Gly Val Tyr 90

Thr Phe Gly Gly Gly Thr 105

Pro Ser Val Phe Ile Phe 120

Thr Ala Ser Val Val Cys 135

Lys Val Gln Trp Lys Val 150 155

Glu Ser Val Thr Glu Gln 170

Ser Thr Leu Thr Leu Ser 185

Ala Cys Glu Val Thr His 200

Phe Asn Arg Gly Glu Cys 215

Pro Val Thr Pro Gly 15

Ser Leu Leu Arg Ser 30

Lys Pro Gly Gln Ser 45

Ala Ser Gly Val Pro 60

Phe Thr Leu Lys Ile 80

Tyr Cys Met Gln Gln 95

Lys Val Glu Ile Lys 110

Pro Pro Ser Asp Glu 125

Leu Leu Asn Asn Phe 140

Asp Asn Ala Leu Gln 160

Asp Ser Lys Asp Ser 175

Lys Ala Asp Tyr Glu 190

Gln Gly Leu Ser Ser 205 <210> 128 <211> 219 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 128

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser

- 180 046146

Ala Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu

100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln

165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly

105

Pro Ser Val Phe Ile

120

Thr Ala Ser Val Val

135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu

170

Ser Thr Leu Thr Leu

185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile

110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 129 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 129

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Met Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu 170 Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser

- 181 046146

210

215 <210> 130 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 130

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Leu Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser Leu 10

Ser Cys Arg Ser Ser Gln 25

Leu Asp Trp Tyr Leu Gln 40

Tyr Leu Gly Ser Asn Arg 55

Ser Gly Ser Gly Thr Asp 70 75

Glu Asp Val Gly Val Tyr 90

Thr Phe Gly Gly Gly Thr 105

Pro Ser Val Phe Ile Phe 120

Thr Ala Ser Val Val Cys 135

Lys Val Gln Trp Lys Val 150 155

Glu Ser Val Thr Glu Gln 170

Ser Thr Leu Thr Leu Ser 185

Ala Cys Glu Val Thr His 200

Phe Asn Arg Gly Glu Cys 215

Pro Val Thr Pro Gly 15

Ser Leu Leu Arg Ser 30

Lys Pro Gly Gln Ser 45

Ala Ser Gly Val Pro 60

Phe Thr Leu Lys Ile 80

Tyr Cys Met Gln Gln 95

Lys Val Glu Ile Lys 110

Pro Pro Ser Asp Glu 125

Leu Leu Asn Asn Phe 140

Asp Asn Ala Leu Gln 160

Asp Ser Lys Asp Ser 175

Lys Ala Asp Tyr Glu 190

Gln Gly Leu Ser Ser 205 <210> 131 <211> 219 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 131

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 1015

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 20 2530

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 4045

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 5560

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu LysIle

70 7580

Ser Arg Val Glu Ala Glu Asp Val Gly Ala Tyr Tyr Cys Met GlnGln

9095

- 182 046146

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val

135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu 170

Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 132 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 132

Asp Ile Val Met Thr 1 5

Gly Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Leu Ser Gly 65 Ser Arg Val Glu Ala 85

Gln Glu Thr Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu 170 Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys <210> 133 <211> 219 <212> PROTEIN <213> Artificial <220>

subsequence

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser

- 183 046146 <223> Synthetic <400> 133

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Thr Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu 170

Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu His 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 134 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 134

Asp Ile Val Met Thr 1 5

Glu Ser Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu

Leu Pro Val Thr Pro 15

Gln Gly Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln

160

Ser

- 184 046146

165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

170175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

185190

Ala Cys Glu Val Thr His Gln Gly Leu Ser

200205

Phe Asn Arg Gly Glu Cys

215

Glu

Ser <210> 135 <211> 452 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 135

Gln Val Gln Leu Gln

5

Thr Leu Ser Leu Thr

Tyr Tyr Trp Gly Trp 35

Ile Gly Thr Ile Tyr 50

Glu Ser Arg Val Thr 65

Leu Lys Leu Ser Ser 85

Ala Arg Gln Gly Ser 100

Gly Gln Gly Thr Thr 115

Ser Val Phe Pro Leu

130

Ala Ala Leu Gly Cys 145

Val Ser Trp Asn Ser 165

Ala Val Leu Gln Ser 180

Val Pro Ser Ser Ser 195

His Lys Pro Ser Asn 210

Cys Asp Lys Thr His 225

Gly Gly Pro Ser Val

245

Met Ile Ser Arg Thr 260

His Glu Asp Pro Glu 275

Val His Asn Ala Lys 290

Tyr Arg Val Val Ser 305

Gly Lys Glu Tyr Lys

325

Ile Glu Lys Thr Ile 340

Val Tyr Thr Leu Pro sequence construct

Glu

Cys

Ile

His

Ile 70

Val

Ile

Val

Ala

Leu 150 Gly

Ser

Leu

Thr

Thr 230 Ph

Pro

Val

Thr

Val 310 Cys

Ser

Pro

Ser Gly Pro

Ala Val Ser 25

Arg Gln Pro 40

Ser Gly Ser 55

Ser Val Asp

Thr Ala Ala

Gln Gln Gly 105

Thr Val Ser 120

Pro Ser Ser 135

Val Lys Asp

Ala Leu Thr

Gly Leu Tyr 185

Gly Thr Gln 200

Lys Val Asp 215

Cys Pro Pro

Leu Phe Pro

Glu Val Thr

265

Lys Phe Asn 280

Lys Pro Arg 295

Leu Thr Val

Lys Val Ser

Lys Ala Lys

345

Ser Arg Asp

Gly Leu Val 10

Gly Tyr Ser

Pro Gly Lys

Thr Tyr Tyr 60

Thr Ser Lys 75

Asp Thr Ala 90

Tyr Tyr Gly

Ser Ala Ser

Lys Ser Thr

140

Tyr Phe Pro 155

Ser Gly Val 170

Ser Leu Ser

Thr Tyr Ile

Lys Lys Val

220

Cys Pro Ala 235

Pro Lys Pro 250

Cys Val Val

Trp Tyr Val

Glu Glu Gln

300

Leu His Gln 315

Asn Lys Ala 330

Gly Gln Pro

Glu Leu Thr

Lys Pro Ser 15

Ile Ser Ser 30

Gly Leu Glu 45

Asn Pro Ser

Asn Gln Phe

Val Tyr Tyr 95

Met Asp Val 110

Thr Lys Gly 125

Ser Gly Gly

Glu Pro Val

His Thr Phe 175

Ser Val Val 190

Cys Asn Val 205

Glu Pro Lys

Pro Glu Ala

Lys Asp Thr 255

Val Asp Val 270

Asp Gly Val 285

Tyr Asn Ser

Asp Trp Leu

Leu Pro Ala

335

Arg Glu Pro 350

Lys Asn Gln

Glu

Gly

Trp

Leu

Ser 80

Cys

Trp

Pro

Thr

Thr 160 Pro

Thr

Asn

Ser

Ala 240 Leu

Ser

Glu

Thr

Asn 320 Ser

Gln

Val

- 185 046146

355 360365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrPro

385 390 395400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys LeuThr

405 410415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440445

Ser Pro Gly Lys

450 <210> 136 <211> 451 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 136

Gln Val Gln Leu Gln 1 5

Thr Leu Ser Leu Thr

Tyr Tyr Trp Gly Trp 35

Ile Gly Thr Ile Tyr 50

Glu Ser Arg Val Thr 65

Leu Lys Leu Ser Ser 85

Ala Arg Gln Gly Ser 100

Gly Gln Gly Thr Thr 115

Ser Val Phe Pro Leu

130

Ala Ala Leu Gly Cys 145

Val Ser Trp Asn Ser

165

Ala Val Leu Gln Ser 180

Val Pro Ser Ser Ser 195

His Lys Pro Ser Asn 210

Cys Asp Lys Thr His 225

Gly Gly Pro Ser Val

245

Met Ile Ser Arg Thr 260

His Glu Asp Pro Glu 275

Val His Asn Ala Lys 290

Tyr Arg Val Val Ser sequence construct

Glu Ser Gly Pro Gly Leu 10

Cys Ala Val Ser Gly Tyr 25

Ile Arg Gln Pro Pro Gly 40

His Ser Gly Ser Thr Tyr 55

Ile Ser Val Asp Thr Ser 70 75

Val Thr Ala Ala Asp Thr 90

Ile Gln Gln Gly Tyr Tyr 105

Val Thr Val Ser Ser Ala 120

Ala Pro Ser Ser Lys Ser 135

Leu Val Lys Asp Tyr Phe 150 155

Gly Ala Leu Thr Ser Gly 170

Ser Gly Leu Tyr Ser Leu 185

Leu Gly Thr Gln Thr Tyr 200

Thr Lys Val Asp Lys Lys 215

Thr Cys Pro Pro Cys Pro 230 235

Phe Leu Phe Pro Pro Lys 250

Pro Glu Val Thr Cys Val 265

Val Lys Phe Asn Trp Tyr 280

Thr Lys Pro Arg Glu Glu 295

Val Leu Thr Val Leu His

Val Lys Pro Ser Glu 15

Ser Ile Ser Ser Gly 30

Lys Gly Leu Glu Trp 45

Tyr Asn Pro Ser Leu 60

Lys Asn Gln Phe Ser 80

Ala Val Tyr Tyr Cys 95

Gly Met Asp Val Trp 110

Ser Thr Lys Gly Pro 125

Thr Ser Gly Gly Thr 140

Pro Glu Pro Val Thr

160

Val His Thr Phe Pro 175

Ser Ser Val Val Thr 190

Ile Cys Asn Val Asn 205

Val Glu Pro Lys Ser 220

Ala Pro Glu Ala Ala

240

Pro Lys Asp Thr Leu 255

Val Val Asp Val Ser 270

Val Asp Gly Val Glu 285

Gln Tyr Asn Ser Thr 300

Gln Asp Trp Leu Asn

- 186 046146

305

Gly Lys Glu Tyr Lys 325

Ile Glu Lys Thr Ile 340

Val Tyr Thr Leu Pro 355

Ser Leu Thr Cys Leu 370

Glu Trp Glu Ser Asn 385

Pro Val Leu Asp Ser 405

Val Asp Lys Ser Arg 420

Met His Glu Ala Leu 435

Ser Pro Gly 450

310 315 Cys Lys Val Ser Asn Lys Ala 330 Ser Lys Ala Lys Gly Gln Pro 345 Pro Ser Arg Asp Glu Leu Thr 360 Val Lys 375 Gly Phe Tyr Pro Ser 380 Gly Gln 390 Pro Glu Asn Asn Tyr 395 Asp Gly Ser Phe Phe Leu Tyr 410 Trp Gln Gln Gly Asn Val Phe 425 His Asn His Tyr Thr Gln Lys 440

320

Leu Pro Ala Ser 335

Arg Glu Pro Gln 350

Lys Asn Gln Val 365

Asp Ile Ala Val

Lys Thr Thr Pro

400

Ser Lys Leu Thr 415

Ser Cys Ser Val 430

Ser Leu Ser Leu 445 <210> 137 <211> 452 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 137

Gln Val Gln Leu Gln 1 5

Thr Leu Ser Leu Thr 20

Tyr Tyr Trp Gly Trp 35

Ile Gly Thr Ile Tyr 50

Lys Ser Arg Val Thr 65

Leu Lys Leu Ser Ser 85

Ala Arg Gln Gly Ser 100

Gly Gln Gly Thr Thr 115

Ser Val Phe Pro Leu 130

Ala Ala Leu Gly Cys 145

Val Ser Trp Asn Ser 165

Ala Val Leu Gln Ser 180

Val Pro Ser Ser Ser

195

His Lys Pro Ser Asn 210

Cys Asp Lys Thr His 225

Gly Gly Pro Ser Val

245

Met Ile Ser Arg Thr sequence construct

Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1015

Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly 2530

Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 4045

His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu 5560

Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 70 7580

Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Ile Lys Gln Gly Tyr Tyr Gly Met Asp Val Trp 105110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 120125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 135140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 150 155160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 170175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 185190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 200205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 215220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 230 235240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 250255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser

- 187 046146

260

His Glu Asp Pro Glu 275

Val His Asn Ala Lys 290

Tyr Arg Val Val Ser 305

Gly Lys Glu Tyr Lys

325

Ile Glu Lys Thr Ile 340

Val Tyr Thr Leu Pro 355

Ser Leu Thr Cys Leu 370

Glu Trp Glu Ser Asn 385

Pro Val Leu Asp Ser

405

Val Asp Lys Ser Arg 420

Met His Glu Ala Leu 435

Ser Pro Gly Lys 450

265

Val Lys Phe Asn Trp Tyr 280

Thr Lys Pro Arg Glu Glu 295

Val Leu Thr Val Leu His 310 315

Cys Lys Val Ser Asn Lys 330

Ser Lys Ala Lys Gly Gln 345

Pro Ser Arg Asp Glu Leu 360

Val Lys Gly Phe Tyr Pro 375

Gly Gln Pro Glu Asn Asn 390 395

Asp Gly Ser Phe Phe Leu 410

Trp Gln Gln Gly Asn Val 425

His Asn His Tyr Thr Gln 440

270

Val Asp Gly Val Glu 285

Gln Tyr Asn Ser Thr 300

Gln Asp Trp Leu Asn

320

Ala Leu Pro Ala Ser

335

Pro Arg Glu Pro Gln 350

Thr Lys Asn Gln Val 365

Ser Asp Ile Ala Val 380

Tyr Lys Thr Thr Pro

400

Tyr Ser Lys Leu Thr 415

Phe Ser Cys Ser Val 430

Lys Ser Leu Ser Leu 445 <210> 138 <211> 451 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 138

Gln Val Gln Leu Gln Glu Ser Gly 15

Thr Leu Ser Leu Thr Cys Ala Val 20

Tyr Tyr Trp Gly Trp Ile Arg Gln 3540

Ile Gly Thr Ile Tyr His Ser Gly 5055

Lys Ser Arg Val Thr Ile Ser Val

6570

Leu Lys Leu Ser Ser Val Thr Ala 85

Ala Arg Gln Gly Ser Ile Lys Gln 100

Gly Gln Gly Thr Thr Val Thr Val 115120

Ser Val Phe Pro Leu Ala Pro Ser 130135

Ala Ala Leu Gly Cys Leu Val Lys 145150

Val Ser Trp Asn Ser Gly Ala Leu 165

Ala Val Leu Gln Ser Ser Gly Leu 180

Val Pro Ser Ser Ser Leu Gly Thr

195200

His Lys Pro Ser Asn Thr Lys Val

Pro Gly Leu Val Lys Pro Ser Glu 1015

Ser Gly Tyr Ser Ile Ser Ser Gly 2530

Pro Pro Gly Lys Gly Leu Glu Trp 45

Ser Thr Tyr Tyr Asn Pro Ser Leu 60

Asp Thr Ser Lys Asn Gln Phe Ser 7580

Ala Asp Thr Ala Val Tyr Tyr Cys 9095

Gly Tyr Tyr Gly Met Asp Val Trp 105110

Ser Ser Ala Ser Thr Lys Gly Pro 125

Ser Lys Ser Thr Ser Gly Gly Thr 140

Asp Tyr Phe Pro Glu Pro Val Thr 155160

Thr Ser Gly Val His Thr Phe Pro 170175

Tyr Ser Leu Ser Ser Val Val Thr 185190

Gln Thr Tyr Ile Cys Asn Val Asn 205

Asp Lys Lys Val Glu Pro Lys Ser

- 188 046146

210

Cys Asp Lys Thr His 225

Gly Gly Pro Ser Val

245

Met Ile Ser Arg Thr 260

His Glu Asp Pro Glu 275

Val His Asn Ala Lys 290

Tyr Arg Val Val Ser 305

Gly Lys Glu Tyr Lys

325

Ile Glu Lys Thr Ile 340

Val Tyr Thr Leu Pro 355

Ser Leu Thr Cys Leu 370

Glu Trp Glu Ser Asn 385

Pro Val Leu Asp Ser

405

Val Asp Lys Ser Arg 420

Met His Glu Ala Leu 435

Ser Pro Gly

450

215

Thr Cys Pro Pro Cys 230

Phe Leu Phe Pro Pro

250

Pro Glu Val Thr Cys 265

Val Lys Phe Asn Trp 280

Thr Lys Pro Arg Glu 295

Val Leu Thr Val Leu 310

Cys Lys Val Ser Asn 330

Ser Lys Ala Lys Gly 345

Pro Ser Arg Asp Glu 360

Val Lys Gly Phe Tyr 375

Gly Gln Pro Glu Asn 390

Asp Gly Ser Phe Phe 410

Trp Gln Gln Gly Asn 425

His Asn His Tyr Thr 440

220

Pro Ala Pro Glu Ala 235

Lys Pro Lys Asp Thr 255

Val Val Val Asp Val 270

Tyr Val Asp Gly Val 285

Glu Gln Tyr Asn Ser 300

His Gln Asp Trp Leu 315

Lys Ala Leu Pro Ala

335

Gln Pro Arg Glu Pro 350

Leu Thr Lys Asn Gln 365

Pro Ser Asp Ile Ala 380

Asn Tyr Lys Thr Thr 395

Leu Tyr Ser Lys Leu 415

Val Phe Ser Cys Ser 430

Gln Lys Ser Leu Ser 445

Ala 240 Leu

Ser

Glu

Thr

Asn 320 Ser

Gln

Val

Val

Pro 400 Thr

Val

Leu <210> 139 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 139

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln

160

Ser

- 189 046146

165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

170175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

185190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

200205

Phe Asn Arg Gly Glu Cys

215 <210> 140 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 140

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Asn Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 4045

Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 5560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 70 7580

Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gln 9095

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 105110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 120125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 135140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 150 155160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 170175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 185190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 200205

Phe Asn Arg Gly Glu Cys 215 <210> 141 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 141

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile

Asn Gly Tyr Asn Tyr 35 sequence construct

Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1015

Ser Cys Arg Ser Ser Gln Ser Leu Leu Arg Ser 2530

Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Pro 4045

- 190 046146

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Val Glu Ala 85

Gln Glu Thr Pro Leu

100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln

165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu

170

Ser Thr Leu Thr Leu

185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Arg Val Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp

175

Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser <210> 142 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 142

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Thr Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser 10

Ser Cys Arg Ser Ser 25

Leu Asp Trp Tyr Leu 40

Tyr Leu Gly Ser Asn 55

Ser Gly Ser Gly Thr 70

Glu Asp Val Gly Val 90

Thr Phe Gly Gly Gly 105

Pro Ser Val Phe Ile 120

Thr Ala Ser Val Val 135

Lys Val Gln Trp Lys 150

Glu Ser Val Thr Glu 170 Ser Thr Leu Thr Leu 185

Ala Cys Glu Val Thr 200

Phe Asn Arg Gly Glu 215

Leu Pro Val Thr Pro 15

Gln Ser Leu Leu Arg 30

Gln Lys Pro Gly Gln 45

Arg Ala Ser Gly Val 60

Asp Phe Thr Leu Lys 75

Tyr Tyr Cys Met Gln 95

Thr Lys Val Glu Ile 110

Phe Pro Pro Ser Asp 125

Cys Leu Leu Asn Asn 140

Val Asp Asn Ala Leu 155

Gln Asp Ser Lys Asp 175 Ser Lys Ala Asp Tyr 190

His Gln Gly Leu Ser 205

Cys

Gly

Ser

Ser

Pro

Ile 80 Gln

Lys

Glu

Phe

Gln 160 Ser

Glu

Ser

- 191 046146 <210> 143 <211> 219 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 143

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 Glu Pro Ala Ser 5 Ile Ser Cys Arg Ser 10 Ser Gln Ser Leu Leu 15 Arg Ser Ser Gly Tyr 20 Asn Tyr Leu Asp Trp 25 Tyr Leu Gln Lys Pro thirty Gly Gln Ser Pro Gln 35 Leu Leu Ile Tyr Leu 40 Gly Ser Asn Arg Ala 45 Ser Gly Val Pro Asp 50 Arg Phe Ser Gly Ser 55 Gly Ser Gly Thr Asp 60 Phe Thr Leu Lys Ile 65 Ser Arg Ala Glu Ala 70 Glu Asp Val Gly Val 75 Tyr Tyr Cys Met Gln 80 Gln Gln Glu Ala Pro 85 Leu Thr Phe Gly Gly 90 Gly Thr Lys Val Glu 95 Ile Lys Arg Thr Val 100 Ala Ala Pro Ser Val 105 Phe Ile Phe Pro Pro 110 Ser Asp Glu Gln Leu 115 Lys Ser Gly Thr Ala 120 Ser Val Val Cys Leu 125 Leu Asn Asn Phe Tyr 130 Pro Arg Glu Ala Lys 135 Val Gln Trp Lys Val 140 Asp Asn Ala Leu Gln 145 Ser Gly Asn Ser Gln 150 Glu Ser Val Thr Glu 155 Gln Asp Ser Lys Asp 160 Ser Thr Tyr Ser Leu 165 Ser Ser Thr Leu Thr 170 Leu Ser Lys Ala Asp 175 Tyr Glu Lys His Lys 180 Val Tyr Ala Cys Glu 185 Val Thr His Gln Gly 190 Leu Ser Ser Pro Val 210 195 Thr Lys Ser Phe Asn 215 200 Arg Gly Glu Cys 205

144

219

PROTEIN

Artificial <210>

<211>

<212>

<213>

<220>

<223>

<400> 144

Asp Ile Val Met 1

Glu Pro Ala Ser

Gly Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro

100

Arg Thr Val Ala

Synthetic sequence construct

Pro

Arg

Thr Gln Ser 5

Ile Ser Cys

Tyr Leu Asp Trp

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Ala Pro Ser Val

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Phe Ile Phe Pro

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110

Pro Ser Asp Glu

- 192 046146

115 120125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 130 135140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala LeuGln

145 150 155160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys AspSer

165 170175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210215 <210> 145 <211> 219 <212> PROTEIN <213> Artificial <220>

<223> Synthetic <400> 145

Asp Ile Val Met Thr 1 5

Glu Pro Ala Ser Ile 20

Arg Gly Tyr Asn Tyr 35

Pro Gln Leu Leu Ile 50

Asp Arg Phe Ser Gly 65

Ser Arg Ala Glu Ala 85

Gln Glu Ala Pro Leu 100

Arg Thr Val Ala Ala 115

Gln Leu Lys Ser Gly 130

Tyr Pro Arg Glu Ala 145

Ser Gly Asn Ser Gln 165

Thr Tyr Ser Leu Ser 180

Lys His Lys Val Tyr 195

Pro Val Thr Lys Ser 210 sequence construct

Gln Ser Pro Leu Ser Leu 10

Ser Cys Arg Ser Ser Gln 25

Leu Asp Trp Tyr Leu Gln 40

Tyr Leu Gly Ser Asn Arg 55

Ser Gly Ser Gly Thr Asp 70 75

Glu Asp Val Gly Val Tyr 90

Thr Phe Gly Gly Gly Thr 105

Pro Ser Val Phe Ile Phe 120

Thr Ala Ser Val Val Cys 135

Lys Val Gln Trp Lys Val 150 155

Glu Ser Val Thr Glu Gln 170

Ser Thr Leu Thr Leu Ser 185

Ala Cys Glu Val Thr His 200

Phe Asn Arg Gly Glu Cys 215

Pro Val Thr Pro Gly 15

Ser Leu Leu Arg Ser 30

Lys Pro Gly Gln Ser 45

Ala Ser Gly Val Pro 60

Phe Thr Leu Lys Ile 80

Tyr Cys Met Gln Gln 95

Lys Val Glu Ile Lys 110

Pro Pro Ser Asp Glu 125

Leu Leu Asn Asn Phe 140

Asp Asn Ala Leu Gln 160

Asp Ser Lys Asp Ser 175

Lys Ala Asp Tyr Glu 190

Gln Gly Leu Ser Ser 205 <210> 146 <211> 219 <212> PROTEIN <213> Artificial sequence <220>

<223> Synthetic construct <400> 146

- 193 046146

Asp Ile Val Met 1

Glu Pro Ala Ser

Asp Gly Tyr Asn 35

Pro Gln Leu Leu 50

Asp Arg Phe Ser 65

Ser Arg Ala Glu

Gln Glu Ala Pro

100

Arg Thr Val Ala 115

Gln Leu Lys Ser 130

Tyr Pro Arg Glu 145

Ser Gly Asn Ser

Thr Tyr Ser Leu

180

Lys His Lys Val 195

Pro Val Thr Lys 210

Thr Gln Ser Pro 5

Ile Ser Cys Arg

Tyr Leu Asp Trp 40

Ile Tyr Leu Gly 55

Gly Ser Gly Ser 70

Ala Glu Asp Val 85

Leu Thr Phe Gly

Ala Pro Ser Val

120

Gly Thr Ala Ser 135

Ala Lys Val Gln 150

Gln Glu Ser Val 165

Ser Ser Thr Leu

Tyr Ala Cys Glu

200

Ser Phe Asn Arg 215

Leu Ser Leu Pro 10

Ser Ser Gln Ser 25

Tyr Leu Gln Lys

Ser Asn Arg Ala 60

Gly Thr Asp Phe 75

Gly Val Tyr Tyr 90

Gly Gly Thr Lys 105

Phe Ile Phe Pro

Val Val Cys Leu

140

Trp Lys Val Asp 155

Thr Glu Gln Asp 170

Thr Leu Ser Lys 185

Val Thr His Gln

Gly Glu Cys

Val Thr Pro Gly 15

Leu Leu Arg Ser 30

Pro Gly Gln Ser 45

Ser Gly Val Pro

Thr Leu Lys Ile 80

Cys Met Gln Gln 95

Val Glu Ile Lys 110

Pro Ser Asp Glu 125

Leu Asn Asn Phe

Asn Ala Leu Gln

160

Ser Lys Asp Ser 175

Ala Asp Tyr Glu 190

Gly Leu Ser Ser 205

Claims (54)

1. An antibody that binds to the protein sortilin, where the antibody contains a heavy chain variable region and a light chain variable region, where: (a) the heavy chain variable region contains HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVRH2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVRH3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variable region contains HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32); or (b) the heavy chain variable region contains HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVRH2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVRH3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variable region contains HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32). 2. Antibody according to claim 1, where the antibody: (a) reduces cell surface levels of sortilin more than the level of reduction caused by an anti-sortilin antibody containing a heavy chain variable region containing the sequence of SEQ ID NO: 56 and a light chain variable region containing the sequence of SEQ ID NO: 79; (b) increases extracellular levels of progranulin more than the level of increase caused by an anti-sortilin antibody containing a heavy chain variable region containing the sequence of SEQ ID NO: 56, and a light chain variable region containing the sequence of SEQ ID NO: 79; (c) inhibits the interaction between sortilin and progranulin more than the level of inhibition caused by an anti-sortilin antibody containing a heavy chain variable region containing the sequence of SEQ ID NO: 56 and a light chain variable region containing the sequence of SEQ ID NO: 79; or - 194 046146 (d) provides any combination of (a)-(c). 3. The antibody of claim 1 or 2, wherein the antibody has a dissociation constant (K _D ) with human sortilin that is greater than 1-fold to about 2.1-fold lower than that of an anti-sortilin antibody containing the severe variable region chain containing the sequence SEQ ID NO: 56, and a light chain variable region containing the sequence SEQ ID NO: 79, where K _D is determined by fluorescence-activated cell sorting (FACS). 4. An antibody according to any one of claims 1 to 3, wherein the antibody has a dissociation constant (K _D ) with human sortilin that ranges from about 5.0E-10 M to about 1.0E-9 M, where KD is determined by FACS method, or approximately 250-500 pM, where KD is determined by biolayer interferometry. 5. An antibody according to any one of claims 1 to 4, wherein the heavy chain variable region comprises HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVRH2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVRH3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variable region contains HVR-L1 containing the amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32). 6. Antibody according to any one of claims 1-4, wherein the heavy chain variable region contains HVR-H1 containing the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), HVRH2 containing the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and HVRH3 containing the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variable region comprises HVR-L1 containing the amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), HVR-L2 containing the amino acid sequence LGSNRAS (SEQ ID NO: 29), and HVR-L3 containing the amino acid sequence MQQQEAPLT (SEQ ID NO: 32). 7. An antibody that binds to the protein sortilin, wherein said antibody contains: (a) a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 57; or (b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 60. 8. The antibody according to any one of claims 1 to 5 and 7, wherein said antibody comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 60. 9. The antibody according to any one of claims 1 to 5 and 7, wherein said antibody comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence of SEQ ID NO: 57. 10. An antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 137 and a light chain containing the amino acid sequence of SEQ ID NO: 142. 11. An antibody that binds to the protein sortilin, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 138 and a light chain containing the amino acid sequence of SEQ ID NO: 142. 12. An antibody according to any one of claims 1 to 9, wherein the antibody belongs to the IgG class, the IgM class or the IgA class. 13. The antibody according to claim 12, where the antibody belongs to the IgG class and has an isotype of IgG1, IgG2, IgG3 or IgG4. 14. Antibody according to claim 13, where: (a) the antibody is of the IgG1 or IgG2 isotype and the Fc region contains an amino acid substitution at position P331S, where the residue position numbering follows the EU numbering system; (b) the antibody is of the IgG1 isotype and the Fc region contains amino acid substitutions at positions L234A, L235A and P331S, where the residue position numbering follows the EU numbering system; (c) the antibody is of the IgG1, IgG2 or IgG4 isotype and the Fc region contains an amino acid substitution at position N297A, where the residue position numbering follows the EU numbering system; or (d) the Fc region contains amino acid substitutions at positions S267E and L328F, where the residue position numbering follows the EU numbering system. 15. The antibody according to claim 14, where the antibody belongs to the IgG1 isotype and the Fc region contains amino acid substitutions at positions L234A, L235A and P331S, - 195 046146 where the numbering of the position of the residues corresponds to the EU numbering system. 16. Antibody according to any one of claims 1 to 15, wherein the sortilin protein is human sortilin protein. 17. The antibody according to claim 16, wherein the human sortilin protein contains the amino acid sequence of SEQ ID NO: 81. 18. An antibody according to any one of claims 1 to 16, wherein the sortilin protein is a wild-type protein, a naturally occurring variant, or a disease-associated variant. 19. An antibody according to any one of claims 1 to 18, wherein the antibody is a human antibody or a monoclonal antibody. 20. A conjugate containing an antibody according to any one of claims 1 to 19, where the antibody is conjugated to a cytotoxic agent. 21. A bispecific antibody comprising an antibody according to any one of claims 1 to 19, wherein the bispecific antibody binds to the sortilin protein and to the second antigen. 22. A multivalent antibody comprising an antibody according to any one of claims 1 to 19, wherein the multivalent antibody binds to sortilin protein and at least a second antigen. 23. The bispecific antibody of claim 21 or the multivalent antibody of claim 22, wherein said second antigen is an antigen that facilitates transport across the blood-brain barrier. 24. The bispecific antibody of claim 23, wherein the second antigen is selected from the group consisting of transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor (LPR)-related proteins 1 and 2 -1 and 2), diphtheria toxin receptor, CRM197, llama single domain antibody, TMEM 30(A), protein transduction domain, TAT, Syn-B, penetratin, polyarginine peptide, angiopep peptide, basigin, Glut1, CD98hc and ANG1005. 25. An antibody fragment according to any one of claims 1 to 9, wherein the antibody fragment binds to the human sortilin protein. 26. The antibody fragment of claim 25, wherein the antibody fragment is a Fab-, Fab'-, Fab'-SH-, F(ab')2-, Fv- or scFv-fragment. 27. An isolated nucleic acid that encodes a protein that binds to sortilin, where the nucleic acid contains a nucleic acid sequence encoding an antibody according to any one of claims 1 to 19, a conjugate antibody according to claim 20, a bispecific antibody according to claims 21 and 23 -24, a multivalent antibody according to any one of claims 22 to 24, or an antibody fragment according to claim 25 or 26. 28. An expression vector containing the nucleic acid according to claim 27. 29. An isolated host cell containing the vector of claim 28. 30. A method for producing an antibody that binds to sortilin, wherein the method comprises culturing a host cell according to claim 29 such that the antibody is produced. 31. The method of claim 30, further comprising isolating an antibody produced by the host cell. 32. An isolated antibody that binds to sortilin, produced by the method of claim 30 or 31. 33. A pharmaceutical composition containing an antibody according to any of claims 1-19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23-24, a multivalent antibody according to any of claims 22-24 or an antibody fragment according to claim 25 or 26. 34. A method of preventing, reducing the risk of, or treating an individual with a disease, disorder, or impairment selected from the group consisting of frontotemporal dementia, Alzheimer's disease, vascular dementia, epileptic seizures, retinal dystrophy, amyotrophic lateral sclerosis, traumatic brain injury, damage spinal cord, dementia, stroke, Parkinson's disease, retinal degeneration, arthritis and osteoarthritis, where the method includes administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1-19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23- 24, a multivalent antibody according to any one of claims 22-24 or an antibody fragment according to claim 25 or 26, or a pharmaceutical composition according to claim 33. 35. The method of claim 34, wherein the disease, disorder or lesion is selected from the group consisting of frontotemporal dementia, Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis. 36. A method of inhibiting one or more of neuroinflammation, axonopathy characterized by short axonal sprouting and pathological branching, microglial activation, and inflammatory response in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1 to 19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23-24, a multivalent antibody according to any of claims 22-24 or an antibody fragment according to claim 25 or 26, or a pharmaceutical composition according to claim 33. 37. A method of enhancing one or more of wound healing, autophagy, and clearance of aggregated proteins in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1 to 19, a conjugate according to claim 20, a bispecific an - 196 046146 antibody according to claims 21 and 23-24, a polyvalent antibody according to any of claims 22-24 or an antibody fragment according to claim 25 or 26 or a pharmaceutical composition according to claim 33. 38. A method of preventing, reducing the risk of, or treating arthritis in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1-19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23-24, a multivalent an antibody according to any one of claims 22-24 or an antibody fragment according to claim 25 or 26 or a pharmaceutical composition according to claim 33. 39. A method of reducing the expression of one or more pro-inflammatory mediators in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1-19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23-24, a multivalent an antibody according to any one of claims 22-24 or an antibody fragment according to claim 25 or 26 or a pharmaceutical composition according to claim 33. 40. The method of claim 39, wherein the one or more pro-inflammatory mediators is selected from the group consisting of IL-6, IL12p70, IL12p40, JL-ίβ, TNF-α, CXCL1, CCL2, CCL3, CCL4 and CCL5. 41. A method of inhibiting the interaction between sortilin expressed on a cell and progranulin, including bringing into contact a cell expressing sortilin and an anti-sortilin antibody according to any one of claims 1 to 19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23 -24, a multivalent antibody according to any one of claims 22-24 or an antibody fragment according to claim 25 or 26 or a pharmaceutical composition according to claim 33. 42. The method according to claim 41, where the cell is in in vitro conditions. 43. The method according to claim 41, where the cell is in in vivo conditions. 44. The method according to any one of claims 41-43, further comprising reducing the level of sortilin expressed on the cell surface. 45. The method according to any one of claims 41-44, wherein the extracellular levels of progranulin are increased. 46. A method of increasing progranulin levels in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1-19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23-24, a multivalent antibody according to any one of 2224 or an antibody fragment according to claim 25 or 26 or a pharmaceutical composition according to claim 33. 47. The method according to claim 46, wherein plasma levels of progranulin are increased. 48. The method according to claim 46, wherein the levels of progranulin in the cerebrospinal fluid are increased. 49. A method of reducing sortilin levels in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of an antibody according to any one of claims 1-19, a conjugate according to claim 20, a bispecific antibody according to claims 21 and 23-24, a multivalent antibody according to any one of claims 22-24 or an antibody fragment according to claim 25 or 26 or a pharmaceutical composition according to claim 33. 50. The method according to claim 49, wherein the levels of sortilin in peripheral blood leukocytes are reduced. 51. The method according to any one of claims 34-40 and 46-50, wherein the individual has one or more mutations in the gene encoding progranulin. 52. The method of claim 51, wherein the individual is heterozygous for one or more loss-of-function mutations in the gene encoding progranulin. 53. The method of claim 51 or 52, wherein the individual has or is at risk of developing frontotemporal dementia, Alzheimer's disease or amyotrophic lateral sclerosis. 54. The method according to any one of claims 34-53, where the method includes the use of two or more antibodies against sortilin.