Use of antibodies against sequence similarity family 19 member A5 in the treatment of neuropathic pain

Abstract

The present disclosure provides a method for treating neuropathic pain (including central neuropathic pain or peripheral neuropathic pain) in a subject, the method comprising administering an antibody that specifically binds to human FAM19A5 and a composition comprising such an antibody. The present disclosure also provides a method for treating neuropathic pain including the symptoms of neuropathic pain and / or the root cause of neuropathic pain by administering an antibody that specifically binds to human FAM19A5. In a specific aspect, the method comprises administering an antibody that specifically binds to human FAM19A5, for example, with a KD of 10 nM or less as measured by methods known in the art, such as ELISA, to soluble human FAM19A5, or with a KD of 10 nM or less. D Binds to membrane-bound human FAM19A5, or both.

Description

[0001] Reference to electronically submitted sequence listings

[0002] The contents of the electronically submitted Sequence Listing in the ASCII text file submitted with this application (Name: 3763_009PC01_SeqListing_ST25.txt; Size: 214,313 bytes; Creation Date: April 22, 2019) are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure provides methods for treating neuropathic pain in a subject (eg, a human) using antibodies or antigen-binding fragments thereof that specifically bind to family of sequence similarity 19, member A5 (FAM19A5), or compositions comprising such antibodies or antigen-binding fragments thereof. Background Art

[0004] Neuropathic pain is a chronic pain caused by damage or disease of the somatosensory system. Colloca L et al., Nat Rev Dis Primers 3:17002 (2017). The somatosensory system allows the perception of touch, pressure, pain, temperature, position, movement and vibration. Somatosensory nerves appear in the skin, muscles, joints and fascia and include thermoreceptors, mechanoreceptors, chemoreceptors, itch receptors and nociceptors, which send signals to the spinal cord and ultimately to the brain for further processing. Damage or disease of the somatosensory nervous system can cause changes and disruptions in the transmission of sensory signals to the spinal cord and brain, leading to chronic pain.

[0005] Neuropathic pain places a heavy burden on patients, society, and healthcare systems. It is estimated that approximately 7-10% of the general population suffers from neuropathic pain. Cruccu G. and Truini A., Pain Ther 6:35-42 (2017). Due to various factors (e.g., an aging population, increasing obesity rates, and increased survival rates of cancer patients receiving interventions that may cause neuropathic pain), the prevalence of neuropathic pain is likely to increase further in the future. Moulin D et al., Pain Res Manag 19:328-335 (2014).

[0006] Because the underlying causes are not always fully understood, the management of neuropathic pain is often complex and remains a challenge. Finnerup NB et al., Lancet Neurol 14:162-173 (2015). Current methods focus solely on treating the symptoms of neuropathic pain with pharmacological agents such as tricyclic antidepressants (e.g., amitriptyline, serotonin-norepinephrine reuptake inhibitors (SNRIs), calcium channel α-2-δ ligands gabapentin and pregabalin) and opioids. Cruccu G and Truini A., Pain Ther 6:35-42 (2017). Such drugs typically have serious side effects and / or limited efficacy for many individuals. Therefore, there is a need for more effective treatment options for neuropathic pain. Summary of the Invention

[0007] Disclosed herein is a method for treating neuropathic pain (including central neuropathic pain or peripheral neuropathic pain) in a subject in need thereof. In some embodiments, the method comprises administering to the subject an antagonist of the family of sequence similarity 19, member A5 (FAM19A5) protein.

[0008] In some embodiments, the neuropathic pain is associated with physical injury, infection, diabetes, cancer therapy, alcoholism, amputation, muscle weakness in the back, legs, buttocks, or face, trigeminal neuralgia, multiple sclerosis, shingles, spinal surgery, or any combination thereof. In some embodiments, the neuropathic pain comprises carpal tunnel syndrome, central pain syndrome, degenerative disc disease, diabetic neuropathy, phantom limb pain, postherpetic neuralgia (shingles), pudendal neuralgia, sciatica, low back pain, trigeminal neuralgia, or any combination thereof.

[0009] In some embodiments, the neuropathic pain is caused by nerve compression. In some embodiments, the diabetic neuropathy is diabetic peripheral neuropathy. In some embodiments, the neuropathic pain is sciatica.

[0010] Also provided herein is a method of increasing the threshold or latency of an external stimulus in a subject in need thereof, comprising administering to the subject an antagonist of an anti-FAM19A5 protein. In some embodiments, the external stimulus is a mechanical stimulus. In other embodiments, the external stimulus is a thermal stimulus.

[0011] The present disclosure also provides a method of increasing sensory nerve conduction velocity in a subject in need thereof, comprising administering to the subject an antagonist against the FAM19A5 protein.

[0012] In some embodiments, the FAM19A5 antagonist is an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA, or a vector comprising the same that specifically targets FAM19A5. In other embodiments, the FAM19A5 antagonist is an anti-FAM19A5 antibody, a polynucleotide encoding an anti-FAM19A5 antibody, a vector comprising the polynucleotide thereof, a cell comprising the polynucleotide thereof, or any combination thereof.

[0013] In some embodiments, the FAM19A5 antagonist is an anti-FAM19A5 antibody.

[0014] In some embodiments, the anti-FAM19A5 antibody exhibits a property selected from the group consisting of: (a) binding to soluble human FAM19A5 with a KD of 10 nM or less as measured by enzyme-linked immunosorbent assay (ELISA); (b) binding to membrane-bound human FAM19A5 with a KD of 10 nM or less as measured by ELISA; or (c) both (a) and (b).

[0015] In some embodiments, the anti-FAM19A5 antibody cross-competes for binding to a human FAM19A5 epitope with a reference antibody comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 17, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 18, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 19, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 29, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 30, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 31; (ii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 15, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: NO:27, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:28; (iii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:11, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:12, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:13, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:23, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:24, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:25; or (iv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:20, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:21, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:22, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:32, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:33, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:34.

[0016] In some embodiments, the anti-FAM19A5 antibody binds to the same FAM19A5 epitope as a reference antibody comprising a heavy chain CDR1, CDR2, and CDR3, and a light chain CDR1, CDR2, and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 17, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 18, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 19, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 29, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 30, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 31; (ii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 15, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: NO:27, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:28; (iii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:11, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:12, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:13, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:23, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:24, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:25; or (iv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:20, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:21, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:22, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:32, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:33, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:34.

[0017] In some embodiments, the anti-FAM19A5 antibody binds to at least one FAM19A5 epitope that is SEQ ID NO: 6 or SEQ ID NO: 9. In other embodiments, the anti-FAM19A5 antibody binds only to a FAM19A5 epitope that is SEQ ID NO: 6 or SEQ ID NO: 9. In certain embodiments, the anti-FAM19A5 antibody further binds to an additional FAM19A5 epitope. In some embodiments, the additional FAM19A5 epitope is selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and any combination thereof.

[0018] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3. In certain embodiments, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 19, SEQ ID NO: 16, SEQ ID NO: 13, or SEQ ID NO: 22. In other embodiments, the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 14, SEQ ID NO: 11, or SEQ ID NO: 20. In some embodiments, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 18, SEQ ID NO: 15, SEQ ID NO: 12, or SEQ ID NO: 21.

[0019] In some embodiments, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 26, SEQ ID NO: 23, or SEQ ID NO: 32. In certain embodiments, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 27, SEQ ID NO: 24, or SEQ ID NO: 33. In some embodiments, the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 31, SEQ ID NO: 28, SEQ ID NO: 25, or SEQ ID NO: 34.

[0020] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein (i) the heavy chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: 17, 18, and 19, respectively, and the light chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: 29, 30, and 31, respectively; (ii) the heavy chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: 14, 15, and 16, respectively, and the light chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: 26, 27, and 28, respectively; (iii) the heavy chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: 11, 12, and 13, respectively, and the light chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: 23, 24, and 25, respectively; or (iv) the heavy chain CDR1, CDR2, and CDR3 comprise SEQ ID NOs: ID NOs: 20, 21 and 22, and the light chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 32, 33 and 34, respectively.

[0021] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 37, SEQ ID NO: 36, SEQ ID NO: 35, or SEQ ID NO: 38 and / or a light chain variable domain comprising SEQ ID NO: 41, SEQ ID NO: 40, SEQ ID NO: 39, or SEQ ID NO: 42. In certain embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 37 and a light chain variable domain comprising SEQ ID NO: 41. In other embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 36 and a light chain variable domain comprising SEQ ID NO: 40. In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 35 and a light chain variable domain comprising SEQ ID NO: 39. In certain embodiments, an anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO:38 and a light chain variable domain comprising SEQ ID NO:42.

[0022] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 37, 36, 35, or 38. In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 41, 40, 39, or 42. In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 37, 36, 35, or 38; and wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 41, 40, 39, or 42.

[0023] In some embodiments, the anti-FAM19A5 antibody comprises Fab, Fab', F(ab')2, Fv, or single-chain Fv (scFv).

[0024] In some embodiments, the anti-FAM19A5 antibody is selected from the group consisting of IgG1, IgG2, IgG3, IgG4, variants thereof, and any combination thereof. In certain embodiments, the anti-FAM19A5 antibody is an IgG2 antibody, an IgG4 antibody, or a combination thereof. In other embodiments, the anti-FAM19A5 antibody comprises an IgG2 / IgG4 isotype antibody.

[0025] In some embodiments, the anti-FAM19A5 antibody further comprises a constant region without Fc function. In some embodiments, the anti-FAM19A5 antibody is a chimeric antibody, a human antibody, or a humanized antibody.

[0026] In some embodiments, the FAM19A5 antagonist is linked to an agent to form an immunoconjugate. In some embodiments, the FAM19A5 antagonist is formulated with a pharmaceutically acceptable carrier. In some embodiments, the FAM19A5 antagonist is administered intravenously, orally, parenterally, intrathecally, intracerebroventricularly, intrapulmonaryly, intramuscularly, subcutaneously, intraperitoneally, intravitreally, or intraventricularly.

[0027] In some embodiments, the subject is a human. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1A 、 1B 1C and 1D show binding analysis of individual scFv clones to FAM19A5 protein. Absorbance was measured at 405 nM. Clone number is shown on the X-axis. Figure 1A 、 1B A and 1C show the analysis of 96 clones from 3rd, 4th or 5th level biopanning of the first, second and third chickens, respectively.

[0029] Figure 2 Shown is a schematic diagram for subcloning anti-FAM19A5 antibody (scFv) into a mammalian expression vector.

[0030] Figure 3 The SDS-PAGE results of the chimeric anti-FAM19A5-IgG2 / 4 monoclonal antibody (1-65) are shown. The left panel shows reducing SDS-PAGE, and the right panel shows non-reducing SDS-PAGE.

[0031] Figure 4A It was shown that the chimeric anti-FAM19A5-IgG2 / 4 monoclonal antibody (1-65) specifically binds to human FAM19A5.

[0032] Figure 4B It was shown that the chimeric anti-FAM19A5-IgG2 / 4 monoclonal antibody (1-65) is specific for the FAM19A5 protein but does not bind to other proteins in the FAM19A subfamily.

[0033] Figure 5A and 5BIt is shown that administering anti-FAM19A5 antibodies alleviates the mechanical hyperalgesia of rats induced by chronic contraction injury (CCI). The impact of mechanical hyperalgesia is shown as a threshold value, under which animals withdraw their paws in response to mechanical stimulation. Before threshold measurement, animals have received one of the following: (i) control antibodies (human IgG) (" G2 "), (ii) anti-FAM19A5 antibodies (" G3 ") or (iii) pregabalin (" G4 "). Initial (healthy) animals are used as " false controls " (that is, without CCI induction and without antibody administration) (" G1 "). Figure 5A Shown are threshold data on day 6 after CCI induction. Figure 5B Threshold data on day 21 after CCI induction are shown. Figure 5A and 5B The bar graphs show the data as mean ± SD. The tables below the graphs show the data from individual mice in each group.

[0034] Figure 6A and 6B It is shown that administering anti-FAM19A5 antibodies improves the motor function of rats induced by CCI. As described in the Examples, the effect on motor function is shown as the latency (seconds) to fall from the rotarod. Before threshold measurement, CCI-induced animals had received one of the following: (i) control antibody (human IgG) ("G2"), (ii) anti-FAM19A5 antibody ("G3") or (iii) pregabalin ("G4"). Initial (healthy) animals were used as "sham controls" (i.e., without CCI induction and without antibody administration) ("G1"). Figure 6A Data are shown for day 6 after CCI induction. Figure 6B Data from day 21 after CCI induction are shown. The bar graph shows latency data as mean ± SD. The table below the graph shows data from individual mice in each group.

[0035] Figure 7 An alignment of the FAM19A5 amino acid sequences of different species (i.e., human, mouse, rat, and chicken) is shown. Fragments F1-F6 used for epitope mapping analysis are shown. The signal peptide is underlined.

[0036] Figure 8 The amino acid sequence of epitopes F1-F6 (conjugated to BS) and their location on the human FAM19A5 polypeptide are shown. The top amino acid sequence shown is wild-type FAM19A5 isoform 2 (without a signal peptide). The second amino acid sequence shown is the same sequence, but the cysteine ​​residues have been mutated to serine to reduce nonspecific activity during peptide synthesis. The sizes of the different epitope fragments are shown in brackets.

[0037] Figure 9 The Western blot results for the epitope mapping of monoclonal antibody clone 1-65 and epitope fragments F1 to F6 (lanes 3-8, respectively) are shown. FAM19A5-Cκ (lane 1), PSA-Cκ (lane 2), peptide NDV-BSA (lane 9) and BSA (lane 10) are used as controls. The respective sizes of the different antigens used are shown on the right side of the blot. The amount of the antigen used in each well is 300 ng. The primary antibody used for Western blot is 1-65-scFv-rabbit-Fc-SSS (2 μg / mL), and the secondary antibody used for the experiment is anti-rabbit IgG (Fc specificity)-HRP (1:4000).

[0038] Figure 10 It is shown that the administration of anti-FAM19A5 antibodies improves the mechanical allodynia of diabetic peripheral neuropathy (DPN) rats. The effect of mechanical allodynia is shown as threshold value (g), at which animals retract their paws (" paw retraction threshold value ") in response to mechanical stimulation. About three weeks after inducing diabetic peripheral neuropathy in rats, animals receive control antibodies (NHI, open squares, n=9) or anti-FAM19A5 antibodies (3-2) (filled squares, n=9) every week. Initial (i.e., healthy, without DPN induction) animals are used as controls (open circles, n=5). Data are expressed as mean ± SD. " * " represents a statistically significant difference (p < 0.05) compared to animals receiving control antibodies.

[0039] Figure 11A and 11B It is shown that administration of anti-FAM19A5 antibodies improves thermal hyperalgesia in diabetic peripheral neuropathy (DPN) rats. The effect on thermal hyperalgesia is shown as the duration before the animals withdraw their paws in response to a thermal stimulus ("paw withdrawal latency"). DPN rats received a control antibody (NHI, squares) or anti-FAM19A5 antibody (3-2) (triangles). Figure 11A Thermal hyperalgesia data are shown 4 weeks after antibody administration. Figure 11B Shown are thermal hyperalgesia data 8 weeks after antibody administration. Initial (ie, healthy, no DPN induction) animals were used as controls. Each symbol represents an individual animal. Mean ± SD is also shown.

[0040] Figure 128 weeks after antibody administration, the administration of anti-FAM19A5 antibodies improved sensory nerve conduction velocity (m / s) in diabetic peripheral neuropathy (DPN) rats. DPN rats received a control antibody (NHI, squares) or anti-FAM19A5 antibody (3-2) (triangles). Initial (i.e., healthy, no DPN induction) animals were used as controls. Each symbol represents an individual animal. Mean ± SD is also shown. DETAILED DESCRIPTION

[0041] Disclosed herein is a method for treating neuropathic pain in a subject in need thereof, comprising administering to the subject an antagonist (e.g., an isolated monoclonal antibody or antigen-binding portion thereof) that specifically binds to human family of sequence similarity 19, member A5 (FAM19A5) protein.

[0042] To facilitate understanding of the disclosure provided herein, various terms and phrases are defined. Additional definitions are set forth throughout the detailed description.

[0043] I. Definition

[0044] Throughout this disclosure, the term "a" or "an" entity refers to one or more of that entity; for example, "antibody" is understood to refer to one or more antibodies. Thus, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein.

[0045] Furthermore, “and / or” as used herein should be taken as a specific disclosure of each of the two specified features or components, with or without the other. Thus, the term “and / or” as used herein in phrases such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0046] It should be understood that wherever aspects are described with the word "comprising," similar aspects described with the term "consisting of" and / or "consisting essentially of" are also provided.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art relevant to the present disclosure. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press provide a general dictionary of many terms used in the present disclosure.

[0048] Units, prefixes and symbols are expressed in the form accepted by the International System of Units (SI). Numerical ranges include numbers that define ranges. Unless otherwise indicated, amino acid sequences are written from left to right in the direction of amino to carboxyl groups. The headings provided herein are not limitations on the various aspects of the disclosure, which can be obtained by reference to the specification as a whole. Therefore, the terms defined immediately below are more completely defined by reference to the entire specification.

[0049] The term "about" is used herein to mean approximately, roughly, roughly, or in the region of... When the term "about" is used in conjunction with a numerical range, it modifies the range by extending the boundaries above and below the numerical values ​​recited. In general, the term "about" can modify a numerical value above and below the recited value by, for example, 10% variance, either upward or downward (higher or lower).

[0050] The term "neuropathic pain" refers to pain at any level affecting the central nervous system (CNS) and / or peripheral nervous system due to injury, damage, and / or inappropriate function. The term "neuropathic pain" includes any and all types of neuropathic pain, regardless of the cause of the neuropathic pain and any and all symptoms.

[0051] Neuropathic pain includes central neuropathic pain and peripheral neuropathic pain. As used herein, the term "central neuropathic pain" refers to pain caused by an obstacle, congenital defect, or damage to the central nervous system (i.e., brain or spinal cord). As used herein, the term "peripheral neuropathic pain" refers to pain caused by damage or infection of the peripheral sensory nerves.

[0052] Symptoms of neuropathic pain can include persistent / chronic pain, spontaneous pain, and allodynia (e.g., a painful response to a stimulus that is not normally painful), hyperalgesia (e.g., an exaggerated response to a painful stimulus that normally causes only mild discomfort, such as a pinprick), hyperesthesia (e.g., an excessive physical sensitivity to stimuli, especially of the skin), or hyperalgesia (e.g., where brief discomfort becomes prolonged, severe pain). In some embodiments, symptoms can be persistent and persist after the primary cause, if present, has been resolved. Merck Manual, Neuropathic Pain, available at merckmanuals.com / professional / neurologic-disorders / pain / neuropathic-pain; Campbell JN and Meyer R A Neuron 52(1):77–92 (2006).

[0053] In some embodiments, types of neuropathic pain may include: (1) neuralgia, (2) deafferentation pain syndrome, (3) complex regional pain syndrome (CRPS), and (4) neuropathy (central or peripheral).

[0054] In certain embodiments, neuropathic pain is neuralgia, which refers to pain radiating along one or more specific nerves (e.g., cranial nerves), usually without any demonstrable pathological changes in the neural structure. Neuralgia includes but is not limited to trigeminal neuralgia (TN), atypical trigeminal neuralgia (ATN), occipital neuralgia, glossopharyngeal neuralgia, postherpetic neuralgia (caused by herpes zoster or herpes), peripheral nerve injury pain, sciatica, lower back pain and atypical facial pain. Chemical irritation, chronic kidney disease, diabetes, inflammation, trauma (including surgery), compression of nerves by nearby structures (e.g., tumors), certain drugs (e.g., cisplatin, paclitaxel or vincristine), porphyria (blood disorder) and infection (e.g., herpes zoster), HIV / AIDS, Lyme disease or syphilis) may all lead to neuralgia.

[0055] In some embodiments, neuropathic pain is a deafferentation pain syndrome, which can be caused by a loss of sensory input from a part of the body (e.g., caused by interruption of peripheral sensory fibers or nerves from the central nervous system). Deafferentation pain syndromes include, but are not limited to, brain injury or spinal cord injury, post-stroke pain, phantom limb pain, paraplegia, brachial plexus avulsion, and lumbar radiculopathy.

[0056] In some embodiments, neuropathic pain is " Complex Regional Pain Syndrome " (CRPS), which is a chronic pain condition that most often affects the arms or legs. In some embodiments, CRPS develops after injury, surgery, stroke, or heart attack. In certain embodiments, CRPS disease is CRPS type I (CRPS-I) (also known as reflex sympathetic dystrophy syndrome). Individuals without confirmed nerve damage are typically classified as CRPS-I types. In other embodiments, CRPS is CRPS type II (CRPS-II) (also known as causalgia), which is associated with confirmed nerve damage.

[0057] In certain embodiments, neuropathic pain is neuropathy, which refers to the pain caused by the function or pathological change (for example, disease or injury) of nerve.Neuropathy is usually characterized by sensory or motor neuron abnormality clinically.In certain embodiments, neuropathy is central neuropathy (for example, the function or pathological change of the central nervous system).In other embodiments, neuropathy is peripheral neuropathy (for example, the function or pathological change of one or more peripheral nerves, including motor nerves, sensory nerves, autonomic nerves or its combination).In certain embodiments, peripheral neuropathy relates to the function or pathological change of a single nerve or nerve group (that is, mononeuropathy).In certain embodiments, peripheral neuropathy relates to the function or pathological change (i.e. polyneuropathy) affecting multiple nerves (local or systemic).In certain embodiments, peripheral neuropathy is roughly the same (i.e. symmetrical polyneuropathy) on both sides of the body.In certain embodiments, peripheral neuropathy affects different areas of health (for example, multiple mononeuritis, multifocal mononeuropathy or multiple mononeuropathy).

[0058] As used herein, "mononeuropathy" is a peripheral neuropathy that relates to the loss of movement or sensation in the area caused by the damage or destruction of a single peripheral nerve or a single nerve group.Mononeuropathy is most often caused by damage or trauma in a local area, for example, it causes long-term pressure / compression on a single nerve. However, some systemic disorders (for example, mononeuritis multiplex) can also cause mononeuropathy. In certain embodiments, the damage or trauma in a local area causes the destruction of the myelin sheath (covering) or part of the nerve cells (axons) of the nerve, which can slow down or prevent the conduction of impulses through the nerve. In certain embodiments, mononeuropathy can affect any part of the body. The example of mononeuropathy pain includes but is not limited to sciatic nerve dysfunction, common peroneal nerve dysfunction, radial nerve dysfunction, ulnar nerve dysfunction, cranial mononeuropathy VI, cranial mononeuropathy VII, cranial mononeuropathy III (compression type), cranial mononeuropathy III (diabetic type), axillary nerve dysfunction, carpal tunnel syndrome, femoral nerve dysfunction, tibial nerve dysfunction, Bell's palsy, thoracic outlet syndrome, carpal tunnel syndrome and sixth (abducens) nerve palsy. Finnerup NB et al., Pain 157(8):1599-1606 (2016); National Institute of Neurological Disorders and Stroke, Peripheral Neuropathy Fact Sheet, available at ninds.nih.gov / disorders / peripheralneuropathy / detail_peripheralneuropathy.htm. In some embodiments, the mononeuropathic pain condition is sciatica.

[0059] As used herein, "polyneuropathy" is a peripheral neuropathy that involves loss of movement or sensation in an area caused by damage or destruction of multiple peripheral nerves. Polyneuropathic pain includes, but is not limited to, post-polio syndrome, post-mastectomy syndrome, diabetic neuropathy, alcoholic neuropathy, amyloid, toxins, AIDS, hypothyroidism, uremia, vitamin deficiency, pain caused by chemotherapy, 2', 3'-dideoxycytidine (ddC) treatment, Guillain-Barré syndrome, or Fabry disease. Finnerup NB et al., Pain 157(8): 1599-1606(2016); National Institute of Neurological Disorders and Stroke, PeripheralNeuropathy Fact Sheet, available at ninds.nih.gov / disorders / peripheralneuropathy / detail_peripheralneuropathy.htm. In some embodiments, polyneuropathy is diabetic peripheral neuropathy. In certain embodiments, diabetic peripheral neuropathy is caused by high blood sugar levels (glucose) and / or high fat levels (eg, triglycerides) in the blood of a diabetic subject, which can cause damage to the patient's peripheral nerves.

[0060] In some embodiments, the peripheral neuropathy disclosed herein can be classified according to the part of the nerve cell (e.g., axon, myelin sheath, or cell body) that is damaged or affected. In some embodiments, the peripheral neuropathy is a distal axonopathy caused by a metabolic or toxic disorder of the axon. In some embodiments, the metabolic disorder includes diabetes, renal failure, deficiency syndromes such as malnutrition and alcoholism. In some embodiments, the metabolic disorder is diabetes, and the distal axonopathy is diabetic neuropathy.

[0061] In some embodiments, peripheral neuropathy is a myelinopathy caused by a primary attack on the myelin sheath or myelinated Schwann cells, resulting in acute failure of impulse conduction. The most common cause is acute inflammatory demyelinating polyneuropathy (AIDP; also known as Guillain-Barré syndrome), but other causes include chronic inflammatory demyelinating syndrome (CIDP), inherited metabolic disorders (e.g., leukodystrophy), or toxins.

[0062] In some embodiments, peripheral neuropathy is a neuropathy caused by the destruction of neurons of the peripheral nervous system (PNS). In some embodiments, the neuropathy is caused by motor neuron disease, sensory neuropathy (e.g., herpes nodularis), toxins, or autonomic dysfunction. In some embodiments, the neuropathy is caused by a neurotoxin (such as the chemotherapeutic agent vincristine).

[0063] Neuropathic pain can be caused by or associated with a variety of etiologies (e.g., physical injury (e.g., trauma or repetitive stress), disease or disorder, exposure to a toxic agent, or a combination thereof). In some embodiments, neuropathic pain is caused by or associated with a traumatic injury or damage, such as, for example, a nerve compression injury (e.g., nerve crush, nerve stretch, nerve entrapment, or incomplete nerve transection); spinal cord injury (e.g., to half of the spinal cord); injury or damage to a peripheral nerve (e.g., motor nerves, sensory nerves, or autonomic nerves, or a combination thereof), amputation; contusion; inflammation (e.g., inflammation of the spinal cord); or surgery. In some embodiments, neuropathic pain is caused by or associated with repetitive stress, including, for example, repetitive, clumsy, and / or forced activities that require prolonged movement of any joint group. Without being bound by any one theory, the resulting irritation can cause inflammation and swelling of ligaments, tendons, and muscles, constricting the narrow passages through which nerves pass (e.g., ulnar neuropathy and carpal tunnel syndrome, which is a neuropathy caused by a restricted or compressed nerve at the elbow or wrist). In some embodiments, neuropathic pain is caused by or is associated with a disease or disorder including, for example, an ischemic event (e.g., a stroke or heart attack), multiple sclerosis, metabolic and / or endocrine diseases or disorders (e.g., diabetes, metabolic diseases, and acromegaly, a condition caused by overproduction of growth hormone and characterized by abnormal enlargement of parts of the skeleton (including joints), leading to nerve entrapment and pain), small vessel disease (e.g., vasculitis, i.e., inflammation of blood vessels) that results in decreased oxygen supply to peripheral nerves (leading to nerve tissue damage), autoimmune diseases (e.g., Sjögren's syndrome, lupus, rheumatoid arthritis, and acute inflammatory demyelinating neuropathy, also known as Guillain-Barré syndrome), renal disorders, cancer or tumors (e.g., neoplastic tumors, neuromas, paraneoplastic syndromes, and other conditions resulting from cancer treatment). In some embodiments, the neuropathic pain is caused by or is associated with an infectious agent (e.g., tick-borne infection, varicella zoster, Epstein-Barr virus, West Nile virus, cytomegalovirus, and herpes simplex virus), acquired immunodeficiency syndrome (AIDS), or infection caused by bacteria (such as Lyme disease, diphtheria, and leprosy), inflammatory disorders, peripheral nerve disorders (e.g., neuromas), hereditary or emerging hereditary disorders (e.g., Charcot-Marie-Tooth disorder includes extreme weakness and wasting of the muscles of the lower leg and foot, gait abnormalities, loss of tendon reflexes, and numbness of the lower limbs), mononeuropathy, or polyneuropathy. In some embodiments, the neuropathic pain is caused by or is associated with an infectious agent (e.g., tick-borne infection, varicella zoster, Epstein-Barr virus, West Nile virus, cytomegalovirus, herpes simplex virus, AIDS).In some embodiments, neuropathic pain is caused by or associated with exposure to toxic agents, including, for example, drugs, alcohol, heavy metals (e.g., lead, arsenic, mercury), industrial agents (e.g., solvents, fumes from glue), or nitrous oxide.

[0064] The term "neuropathic pain associated with a disease or disorder" refers to neuropathic pain that accompanies, is caused by, or results from a disease or disorder (eg, those disclosed herein).

[0065] As used herein, the terms "treat," "treating," and "treatment" refer to any type of intervention or process performed on a subject, or the administration of an active agent to a subject, with the purpose of reversing, alleviating, ameliorating, inhibiting, slowing, or preventing the progression, development, severity, or recurrence of symptoms, complications, conditions, or biochemical markers associated with a disease. Treatment can be for a subject with a disease or a subject without the disease (e.g., for prevention).

[0066] As used herein, "administering" refers to the physical introduction of a therapeutic agent or a composition comprising a therapeutic agent into a subject using any of the various methods and delivery systems known to those skilled in the art. Preferred routes of administration for antibodies described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, intraspinal, intravitreal, or other parenteral routes of administration, such as by injection or infusion. As used herein, the phrase "parenteral administration" refers to modes of administration other than enteral and topical administration, typically by injection, and includes but is not limited to intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardial, intradermal, intratracheal, subcutaneous, subcutaneous, intravitreal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. Alternatively, antibodies as described herein can be administered by non-parenteral routes, such as topical, epidermal, or mucosal routes of administration, such as intranasal, oral, vaginal, rectal, sublingual, or topical administration. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0067] As used herein, the term "therapeutically effective amount" refers to an amount of a drug, alone or in combination with another therapeutic agent, that effectively "treats" a disease or disorder in a subject or reduces the risk, potential, likelihood, or incidence of a disease or disorder (e.g., neuropathic pain). A "therapeutically effective amount" includes an amount of a drug or therapeutic agent that provides some improvement or benefit to a subject suffering from or at risk of a disease or disorder (e.g., neuropathic pain disclosed herein). Thus, a "therapeutically effective" amount refers to an amount that reduces the risk, potential, likelihood, or incidence of a disease or disorder, or alleviates, mitigates, and / or reduces at least one indicator (e.g., neuropathic pain), and / or alleviates at least one clinical symptom of a disease or disorder.

[0068] As used herein, the term "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0069] The term "family of sequence similarity 19 member A5" or "FAM19A5" refers to a protein that belongs to the TAFA family of five highly homologous proteins (also known as the FAM19 family) and is primarily expressed in the brain and spinal cord. FAM19A5 is also known as TAFA5 or chemokine-like protein TAFA-5.

[0070] In humans, the gene encoding FAM19A5 is located on chromosome 22. There are multiple human FAM19A5 (UniProt: Q7Z5A7) isoforms, which are thought to be produced by alternative splicing: isoform 1 (UniProt: Q7Z5A7-1), which consists of 132 amino acids; isoform 2 (UniProt: Q7Z5A7-2), which consists of 125 amino acids; and isoform 3 (UniProt: Q7Z5A7-3), which consists of 53 amino acids. The human FAM19A5 protein is thought to exist in membrane-bound and soluble (secreted) forms. Isoform 1 is thought to be membrane-bound with one transmembrane region. Isoform 2, reported as a secreted protein (soluble) in Tang TY et al., Genomics 83(4):727-34 (2004), contains a signal peptide at amino acid positions 1-25. Isoform 1 is thought to be a membrane protein. Below are the amino acid sequences of the three known human FAM19A5 isoforms.

[0071] (I) Isoform 1 (UniProt: Q7Z5A7-1, transmembrane protein): This isoform has been selected as the canonical sequence.

[0072] MAPSPRTGSR QDATALPSMS STFWAFMILA SLLIAYCSQL AAGTCEIVTL DRDSSQPRRTIARQTARCAC RKGQIAGTTR ARPACVDARI IKTKQWCDML PCLEGEGCDL LINRSGWTCT QPGGRIKTTTVS(SEQ ID NO:1)

[0073] (II) Isoform 2 (UniProt: Q7Z5A7-2, soluble protein):

[0074] MQLLKALWAL AGAALCCFLV LVIHAQFLKE GQLAAGTCEI VTLDRDSSQP RRTIARQTARCACRKGQIAG TTRARPACVD ARIIKTKQWC DMLPCLEGEG CDLLINRSGW TCTQPGGRIK TTTVS(SEQID NO:2)

[0075] (III) Isoform 3 (UniProt: Q7Z5A7-3):

[0076] MYHHREWPAR IIKTKQWCDM LPCLEGEGCD LLINRSGWTC TQPGGRIKTT TVS(SEQ ID NO:3)

[0077] The term "FAM19A5" includes any variant or isoform of FAM19A5 naturally expressed by cells. Thus, the antibodies described herein may cross-react with different isoforms in the same species (e.g., different isoforms of human FAM19A5), or cross-react with FAM19A5 from species other than humans (e.g., mouse FAM19A5). Alternatively, the antibody may be specific for human FAM19A5 and may not exhibit any cross-reactivity with other species. FAM19A5 or any of its variants and isoforms may be isolated or recombinantly produced from cells or tissues that naturally express them. The polynucleotide encoding human FAM19A5 has GenBank accession number BC039396 and the following sequence:

[0078] Table 1 Polynucleotide sequence of human FAM19A5

[0079]

[0080] The term "antagonist against FAM19A5 protein" refers to all antagonists that inhibit the expression of FAM19A5 protein. Such antagonists can be peptides, nucleic acids, or compounds. More specifically, the antagonist can be an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA (peptide nucleic acid) targeting FAM19A5, or a vector containing the same. In some embodiments, the antagonist can be an antibody or antigen-binding portion thereof that specifically binds to the FAM19A5 protein.

[0081] The terms "antibody" and "antibodies" are terms of the art and can be used interchangeably herein and refer to molecules with an antigen binding site that specifically binds to an antigen. As used herein, the terms include complete antibodies and any antigen binding fragment (i.e., "antigen binding portion") or a single chain thereof. In some embodiments, an "antibody" refers to a glycoprotein comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds, or an antigen binding portion thereof. In other embodiments, an "antibody" refers to a single-chain antibody comprising a single variable domain, such as a VHH domain. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. In some naturally occurring antibodies, the heavy chain constant region is composed of three domains, CH1, CH2, and CH3. In some naturally occurring antibodies, each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is composed of one domain, CL.

[0082] The VH and VL regions can be further subdivided into hypervariable regions, known as complementarity determining regions (CDRs), interspersed with more conserved regions, known as framework regions. Each VH and VL is composed of three CDRs and four FRs, arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of antibodies can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0083] The term "Kabat numbering" and similar terms are generally recognized in the art and refer to a system in which the amino acid residues in the heavy chain and light chain variable region of an antibody or its antigen-binding portion thereof are numbered. In some aspects, the CDR of an antibody can be determined according to the Kabat numbering system (see, e.g., Kabat EA&Wu TT (1971) Ann NY Acad Sci 190:382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No.91-3242). Using the Kabat numbering system, the CDR in the antibody heavy chain molecule is generally present in amino acid positions 31 to 35, which optionally can include one or two other amino acids after 35 (referred to as 35A and 35B in the Kabat numbering scheme) (CDR1), amino acid positions 50 to 65 (CDR2) and amino acid positions 95 to 102 (CDR3). Using the Kabat numbering system, the CDRs within an antibody light chain molecule are typically present at amino acid positions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), and amino acid positions 89 to 97 (CDR3). In specific embodiments, the CDRs of the antibodies described herein are determined according to the Kabat numbering scheme.

[0084] The phrases "amino acid position numbering as in Kabat," "Kabat position," and grammatical variants thereof refer to the numbering system for heavy chain variable domains or light chain variable domains in the antibody compilation in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening or insertion of the FW or CDR of the variable domain. For example, a heavy chain variable domain may include a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and inserted residues after heavy chain FW residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). See Table 1B.

[0085] Table 1B

[0086]

[0087]

[0088] For a given antibody, the Kabat numbering of the residues can be determined by aligning the homology regions of the antibody's sequence with the sequence of the "standard" Kabat numbering. Chothia refers to the position of the structural loops (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987)). When numbering using the Kabat numbering convention, the end of the Chothia CDR-H1 loop varies between H32 and H34, depending on the length of the loop (this is because the Kabat numbering scheme places the insertion point at H35A and H35B; if both 35A and 35B are absent, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable region represents a compromise between the Kabat CDRs and the Chothia structural loops and is used by Oxford Molecular's AbM antibody modeling software.

[0089] IMGT (ImMunoGeneTics) also provides a numbering system for immunoglobulin variable regions (including CDRs). See, for example, Lefranc, MP et al., Dev.Comp.Immunol.27:55-77 (2003), which is incorporated herein by reference. The IMGT numbering system is based on the alignment of more than 5,000 sequences, structural data, and characterization of hypervariable loops, and allows easy comparison of variable regions and CDR regions across all species. According to the IMGT numbering scheme, VH-CDR1 is located at positions 26 to 35, VH-CDR2 is located at positions 51 to 57, VH-CDR3 is located at positions 93 to 102, VL-CDR1 is located at positions 27 to 32, VL-CDR2 is located at positions 50 to 52, and VL-CDR3 is located at positions 89 to 97.

[0090] For all heavy chain constant region amino acid positions discussed in this disclosure, numbering is according to the EU index first described in Edelman et al., 1969, Proc. Natl. Acad. Sci. USA 63(1):78-85, which described the amino acid sequence of the first human IgG1 sequenced myeloma protein EU. The EU index of Edelman et al. was also described in Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed., United States Public Health Service, National Institutes of Health, Bethesda. Thus, the phrases "EU index as described in Kabat" or "EU index of Kabat" and "position according to the EU index as described in Kabat" and grammatical variants thereof refer to the residue numbering system for the human IgG1 EU antibody based on Edelman et al., as described in Kabat 1991.

[0091] The numbering system for variable domains (heavy and light chains) and light chain constant region amino acid sequences is described in Kabat 1991.

[0092] The antibody can be any type of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any species (e.g., IgD, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., human IgG1, IgG2, IgG3, and IgG4, and mouse IgG1, IgG2a, IgG2b, and IgG3). Immunoglobulins, such as IgG1, exist in several allotypes that differ from each other in at most a few amino acids. The antibodies disclosed herein can be from any known isotype, class, subclass, or allotype. In certain embodiments, the antibodies described herein are IgG1, IgG2, IgG3, or IgG4 subclasses, or any hybrid thereof. In certain embodiments, the antibodies are human IgG1 subclass, human IgG2 subclass, or human IgG4 subclass.

[0093] "Antibody" includes, for example, naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human and non-human antibodies; fully synthetic antibodies; single-chain antibodies; monospecific antibodies; multispecific antibodies (including bispecific antibodies); tetrameric antibodies comprising two heavy chain and two light chain molecules; antibody light chain monomers; antibody heavy chain monomers; antibody light chain dimers, antibody heavy chain dimers; antibody light chain-antibody heavy chain pairs; intrabodies; heterotypic conjugate antibodies; monovalent antibodies; single-chain antibodies; camelized antibodies; affybodies; anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies) and single-domain antibodies (sdAbs), which include binding molecules composed of a single monomeric variable antibody domain (e.g., VH domain or VL domain) that is fully capable of binding antigen. Harmen MM and Haard HJ Appl Microbiol Biotechnol. 77(1):13-22 (2007)).

[0094] As used herein, the terms "antigen-binding portion" and "antigen-binding fragment" of an antibody are interchangeable and refer to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., human FAM19A5). Such a "fragment" may have a length of, for example, between about 8 and about 1500 amino acids, suitably between about 8 and about 745 amino acids, suitably between about 8 and about 300 amino acids, such as about 8 to about 200 amino acids, or about 10 to about 50 or 100 amino acids. It has been shown that the antigen-binding function of an antibody can be achieved by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody, e.g., an anti-FAM19A5 antibody described herein, include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, and a disulfide-linked Fv (SDFv); (v) a dAb fragment consisting of a VH domain (Ward et al., (1989) Nature 341:544-546); and (vi) an isolated complementarity determining region (CDR) or (vii) a combination of two or more isolated CDRs, optionally linked by a synthetic linker. In addition, although the two domains VL and VH of the Fv fragment are encoded by different genes, they can be connected using recombinant methods through synthetic linkers, enabling them to be formed into a single protein chain, wherein the VL and VH regions are paired to form a monovalent molecule (referred to as single-chain Fv (ScFv); see, e.g., Bird et al., (1988) Science 242:423-426; and Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-chain antibodies are also intended to be included in the "antigen-binding portion" of the term antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the practicality of the fragments is screened in the same manner as complete antibodies. The antigen-binding portion can be produced by recombinant DNA technology, or by enzymatic or chemical cleavage of complete immunoglobulins.

[0095] As used herein, the terms "variable region" and "variable domain" are used interchangeably and are common in the art. A variable region generally refers to a portion of an antibody, typically a portion of a light chain or a heavy chain, typically the amino-terminal portion of about 110 to 120 amino acids in a mature heavy chain and about 90 to 115 amino acids in a mature light chain, which differ greatly in sequence between antibodies and are used for the binding and specificity of a particular antibody to its specific antigen. The variability of the sequence is concentrated in those regions that are referred to as complementary determining regions (CDRs), while the more highly conserved regions in the variable domains are referred to as framework regions (FRs).

[0096] Without wishing to be bound by any particular mechanism or theory, the CDRs of light and heavy chains are considered to be primarily responsible for the interaction and specificity of the antibody with the antigen. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region includes rodent or mouse CDRs and human framework regions (FRs). In a specific embodiment, the variable region is a primate (e.g., non-human primate) variable region. In certain embodiments, the variable region includes rodent or mouse CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0097] As used herein, the term "heavy chain" (HC) when used in reference to antibodies can refer to any of the different types, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the constant domain, which gives rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes, respectively, including the subclasses of IgG, such as IgG1, IgG2, IgG3, and IgG4.

[0098] As used herein, the term "light chain" (LC) when referring to an antibody can refer to any of the different types, such as kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domain. Light chain amino acid sequences are well known in the art. In a specific embodiment, the light chain is a human light chain.

[0099] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0100] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0101] As used herein, the terms "constant region" and "constant domain" are interchangeable and have their ordinary meaning in the art. The constant region is the portion of an antibody, such as the carboxyl terminal portion of a light chain and / or a heavy chain, which is not directly involved in binding the antibody to an antigen but may exhibit various effector functions, such as interactions with Fc receptors. The constant region of an immunoglobulin molecule generally has a more conserved amino acid sequence than the immunoglobulin variable domain.

[0102] "Fc region" (fragment crystallizable region) or "Fc domain" or "Fc" refers to the C-terminal region of an antibody's heavy chain, which mediates the binding of the immunoglobulin to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells) or binding to the first component (C1q) of the classical complement system. Thus, the Fc region includes the constant region of the antibody, excluding the first constant region immunoglobulin domain (e.g., CH1 or CL). In IgG, IgA, and IgD antibody isotypes, the Fc region contains two identical protein fragments derived from the second (CH2) and third (CH3) constant domains of the antibody's two heavy chains; IgM and IgE Fc regions contain three heavy chain constant domains (CH domains 2-4) in each polypeptide chain. For IgG, the Fc region includes immunoglobulin domains Cγ2 and Cγ3 and the hinge between Cγ1 and Cγ2. Although the boundaries of the Fc region of an immunoglobulin heavy chain may vary, the human IgG heavy chain Fc region is generally defined as extending from an amino acid residue at position C226 or P230 (or an amino acid between these two amino acids) to the carboxyl terminus of the heavy chain, with numbering being according to the EU index as in Kabat. The CH2 domain of the human IgG Fc region extends from approximately amino acid 231 to approximately amino acid 340, while the CH3 domain is located on the C-terminal side of the Cm domain in the Fc region, i.e., it extends from approximately amino acid 341 to approximately amino acid 447 of IgG. As used herein, the Fc region can be a native sequence Fc, including any allotypic variant, or a variant Fc (e.g., a non-naturally occurring Fc). Fc can also refer to this region in the context of an isolated or Fc-containing protein polypeptide, such as a "binding protein comprising an Fc region," also referred to as an "Fc fusion protein" (e.g., an antibody or immunoadhesin).

[0103] A "native sequence Fc region" or "native sequence Fc" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include native sequence human IgG1 Fc region; native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region, as well as naturally occurring variants thereof. Native sequence Fc includes various allotypes of Fcs (see, e.g., Jefferis et al. (2009) mAbs 1:1; Vidarsson G et al. Front Immunol. 5:520 (published online October 20, 2014)).

[0104] "Fc receptor" or "FcR" is a receptor that binds to the Fc region of an immunoglobulin. The FcRs that bind to IgG antibodies include receptors of the FcγR family, including allelic variants and alternatively spliced ​​forms of these receptors. The FcγR family consists of three activating (FcγRI, FcγRIII, and FcγRIV in mice; FcγRIA, FcγRIIA, and FcγRIIIA in humans) and one inhibitory (FcγRIIB) receptor. Human IgG1 binds to most human Fc receptors and elicits the strongest Fc effector functions. In terms of the type of activating Fc receptor it binds, it is considered equivalent to mouse IgG2a. In contrast, human IgG4 elicits the least Fc effector functions. Vidarsson G et al., Front Immunol. 5:520 (published online October 20, 2014).

[0105] The constant region can be operated by, for example, recombinant technology to eliminate one or more effector functions. "Effecter function" refers to the interaction of an antibody Fc region with an Fc receptor or ligand, or the biochemical events resulting therefrom. Exemplary "effector functions" include C1q binding, complement dependent cytotoxicity (CDC), Fc receptor binding, FcγR-mediated effector functions such as ADCC and antibody-dependent cell-mediated phagocytosis (ADCP), and down-regulation of cell surface receptors (e.g., B cell receptor; BCR). Such effector functions generally require that the Fc region be combined with a binding domain (e.g., an antibody variable domain). Therefore, the term "constant region without Fc function" includes a constant region that has reduced or does not have one or more effector functions mediated by the Fc region.

[0106] The effector functions of antibodies can be reduced or avoided by different methods. By using antibody fragments lacking the Fc region (e.g., such as Fab, F(ab')2, single-chain Fv (scFv), or sdAbs consisting of monomeric VH or VL domains), the effector functions of antibodies can be reduced or avoided. Alternatively, so-called glycosylated antibodies can be produced by removing sugars attached to specific residues in the Fc region to reduce the effector functions of antibodies while retaining other valuable properties of the Fc region (e.g., extended half-life and heterodimerization). Glycosylated antibodies can be produced by, for example, deleting or changing the residues to which sugars are attached, enzymatically removing sugars, producing antibodies in cells cultured in the presence of glycosylation inhibitors, or by expressing antibodies in cells that cannot glycosylate proteins (e.g., bacterial host cells). See, for example, U.S. Patent Publication No. 20120100140. Another method is to use an Fc region from an IgG subclass with reduced effector functions, for example, IgG2 and IgG4 antibodies are characterized in that they have lower levels of Fc effector functions than IgG1 and IgG3. The residues closest to the hinge region in the CH2 domain of the Fc portion are responsible for the effector function of the antibody because it contains largely overlapping binding sites for C1q (complement) and IgG-Fc receptors (FcγR) on effector cells of the innate immune system. Vidarsson G et al., Front Immunol. 5: 520 (published online on October 20, 2014). Therefore, antibodies with reduced or no Fc effector function can be prepared by: generating, for example, a chimeric Fc region comprising a CH2 domain from an IgG antibody of the IgG4 isotype and a CH3 domain from an IgG antibody of the IgG1 isotype; or a chimeric Fc region comprising a hinge region from IgG2 and a CH2 region from IgG4 (see, e.g., Lau C et al., J. Immunol. 191: 4769-4777 (2013)); or an Fc region with a mutation that results in altered Fc effector function (e.g., reduced or no Fc function). Such Fc regions with mutations are known in the art. See, for example, U.S. Patent Publication No. 20120100140 and the U.S. and PCT applications cited therein, and An et al., mAbs 1:6, 572-579 (2009); the disclosures of which are incorporated herein by reference in their entirety.

[0107] "Hinge", "hinge domain" or "hinge region" or "antibody hinge region" refers to the domain of the heavy chain constant region that connects the CH1 domain to the CH2 domain, and includes the upper, middle and lower parts of the hinge (Roux et al., J. Immunol. 1998 161: 4083). The hinge provides varying degrees of flexibility between the binding region and the effector region of the antibody, and also provides a site for intermolecular disulfide bonding between the two heavy chain constant regions. As used herein, for all IgG isotypes, the hinge begins at Glu216 and ends at Gly 237 (Roux et al., 1998 J Immunol 161: 4083). The sequences of wild-type IgG1, IgG2, IgG3 and IgG4 hinges are known in the art. See, e.g., Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; Vidarsson G et al., Front Immunol. 5:520 (published online Oct. 20, 2014).

[0108] The term "CH1 domain" refers to the heavy chain constant region that connects the variable domain to the hinge in the heavy chain constant domain. As used herein, the CH1 domain starts at A118 and ends at V215. The term "CH1 domain" includes wild-type CH1 domains and naturally occurring variants thereof (e.g., allotypes). The CH1 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known in the art. See, e.g., Kabat EA et al., (1991) supra and Vidarsson G et al., Front Immunol. 5: 520 (published online on October 20, 2014). Exemplary CH1 domains include CH1 domains having mutations that change the biological activity (e.g., half-life) of the antibody, such as described in U.S. Patent Publication No. 20120100140 and the U.S. Patents cited therein and disclosed and PCT disclosed.

[0109] The term "CH2 domain" refers to the heavy chain constant region that connects the hinge to the CH3 domain in the heavy chain constant domain. As used herein, the CH2 domain begins at P238 and ends at K340. The term "CH2 domain" includes wild-type CH2 domains and naturally occurring variants thereof (e.g., allotypes). The CH2 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known in the art. See, for example, Kabat EA et al., (1991) supra and Vidarsson G et al., Front Immunol. 5: 520 (published online on October 20, 2014). Exemplary CH2 domains include CH2 domains with mutations that alter the biological activity of the antibody (e.g., half-life and / or reduced Fc effector function), such as those described in U.S. Patent Publication No. 20120100140 and the U.S. Patents and publications cited therein and PCT publications.

[0110] The term "CH3 domain" refers to the heavy chain constant region that is the C-terminal end of the CH2 domain in the heavy chain constant domain. As used herein, the CH3 domain begins at G341 and ends at K447. The term "CH3 domain" includes wild-type CH3 domains and naturally occurring variants thereof (e.g., allotypes). The CH3 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known in the art. See, for example, Kabat EA et al., (1991) supra and Vidarsson G et al., Front Immunol. 5: 520 (published online on October 20, 2014). Exemplary CH3 domains include CH3 domains with mutations that alter the biological activity (e.g., half-life) of the antibody, such as those described in U.S. Patent Publication No. 20120100140 and the U.S. Patents and publications cited therein, and PCT publications.

[0111] As used herein, "isotype" refers to the antibody class encoded by the heavy chain constant region genes (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibodies).

[0112] "Isotype" refers to naturally occurring variants within a particular isotype group that differ in a few amino acids (see, e.g., Jefferis et al., (2009) mAbs 1:1). The antibodies described herein can be of any allotype. IgG1, IgG2, IgG3, and IgG4 allotypes are known in the art. See, e.g., Kabat EA et al., (1991) supra; Vidarsson G et al., Front Immunol. 5:520 (published online October 20, 2014); and Lefranc MP, mAbs 1:4, 1-7 (2009).

[0113] The phrases "an antibody that recognizes an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody that specifically binds to an antigen."

[0114] As used herein, an "isolated antibody" is intended to be substantially free of other antibodies with different antigenic specificities (e.g., an isolated antibody that specifically binds to FAM19A5 is substantially free of antibodies that specifically bind to antigens other than FAM19A5). However, an isolated antibody that specifically binds to an epitope of FAM19A5 may have cross-reactivity with other FAM19A5 proteins from different species.

[0115] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, "binding affinity" as used herein refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be represented by a dissociation constant (KD). Affinity can be measured and / or expressed in a variety of ways known in the art, including, but not limited to, the equilibrium dissociation constant (KD) and the equilibrium association constant (KA). KD is calculated by the quotient of koff / kon and is expressed as a molar concentration (M), while KA is calculated by the quotient of kon / koff. kon refers to, for example, the association rate constant of an antibody to an antigen, and koff refers to, for example, the dissociation of an antibody from an antigen. kon and koff can be determined by techniques known to those of ordinary skill in the art, such as immunoassays (e.g., enzyme-linked immunosorbent assay (ELISA)), or kinetic exclusion assay (KinExA).

[0116] As used herein, the terms "specifically binds," "specifically recognizes," "specifically binds," "selectively binds," and "selectively binds" are analogous terms in the context of antibodies and refer to molecules (e.g., antibodies) that bind to an antigen (e.g., an epitope or an immune complex), as such binding is understood by those skilled in the art. For example, a molecule that specifically binds to an antigen can typically be detected, for example, by immunoassays, In certain embodiments, the molecule specifically binds to the antigen with a KA that is at least 2 log, 2.5 log, 3 log, 4 log, or greater than the KA when the molecule is bound to another antigen.

[0117] Antibodies are typically bound to their cognate antigens with a high affinity as reflected by a dissociation constant (KD) of 10-5 to 10-11 M or less. Any KD greater than about 10-4 M is generally considered to be non-specific binding. As used herein, "specifically bound" to an antigen refers to an antibody that is bound to an antigen and substantially the same antigen with high affinity, meaning that when determined by, for example, immunoassays (e.g., ELISA) or surface plasmon resonance (SPR) technology in a BIACORE 2000 instrument using a predetermined antigen, there is 10-7 M or less, preferably 10-8 M or less, even more preferably 10-9 M or less, and most preferably between 10-8 M and 10-10 M or less, but not bound to an unrelated antigen with high affinity.

[0118] As used herein, the term "antigen" refers to any natural or synthetic immunogenic substance, such as a protein, peptide or hapten. The antigen may be FAM19A5 or a fragment thereof.

[0119] As used herein, " epi-position " is the term in this area, and refers to the local region of the antigen that antibody can specifically combine.Epi-position can be the continuous amino acid (linear or continuous epi-position) of such as polypeptide, or epi-position can be such as from two or more discontinuous regions (conformation, non-linear, discontinuous or discontinuous epi-position) of one or more polypeptide.The epi-position formed by continuous amino acid is usually but not always retained when being exposed to denaturing solvent, and the epi-position formed by tertiary folding is usually lost when being treated with denaturing solvent.Epi-position generally includes at least 3,4,5,6,7,8,9,10,11,12,13,14,15 or 20 amino acid of unique spatial conformation.The method for determining the epi-position that given antibody is combined (i.e. epi-position mapping) is well-known in the art, and includes such as immunoblotting and immunoprecipitation assay, wherein test is from (for example, from FMAM19A5) overlapping or adjacent peptide and the reactivity of given antibody (for example, anti-FAM19A5 antibody). Methods for determining the spatial conformation of epitopes include techniques in the art and those described herein, e.g., x-ray crystallography, two-dimensional nuclear magnetic resonance, and HDX-MS (see, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, G.E. Morris, Ed. (1996)).

[0120] In certain embodiments, the epitope to which the antibody is bound can be determined by, for example, NMR spectroscopy, X-ray crystallography, ELISA assays, hydrogen / deuterium exchange in combination with mass spectrometry (e.g., liquid chromatography electrospray mass spectrometry), array-based oligopeptide scanning assays and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography, any method known in the art can be used to complete crystallization (e.g., Giege R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem 189: 1-23; Chayen NE (1997) Structure 5: 1269-1274; McPherson A (1976) J Biol Chem 251: 6300-6303). Antibody: antigen crystals can be studied using well-known X-ray diffraction techniques and can be refined using computer software such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.; see, e.g., Meth Enzymol (1985) volumes 114 & 115, eds Wyckoff HW et al.; US 2004 / 0014194), and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49 (Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56 (Pt 10): 1316-1323). Mutagenesis mapping studies can be performed using any method known to those skilled in the art. See, eg, Champe M et al. (1995) J Biol Chem 270: 1388-1394 and Cunningham BC & Wells JA (1989) Science 244: 1081-1085 for a description of mutagenesis techniques including alanine scanning mutagenesis.

[0121] The term "epitope mapping" refers to the process of identifying molecular determinants for antibody-antigen recognition.

[0122] The term "binding to the same epitope" referring to two or more antibodies refers to the same segment of amino acid residues as determined by a given method. The technology for determining whether an antibody is bound to the "same epitope on FAM19A5" as an antibody as described herein includes, for example, epitope mapping methods, such as X-ray analysis (which provides atomic resolution of the epitope) and hydrogen / deuterium exchange mass spectrometry (HDX-MS) of crystals to antigen: antibody complexes. Other methods monitor the binding of antibodies to antigen fragments or mutant variants of antigens, in which the absence of binding caused by the modification of amino acid residues in the antigen sequence is generally considered to be an indication of epitope composition. In addition, a computational combination method for epitope mapping can also be used. These methods rely on the ability of the antibody of interest to affinity separate specific short peptides from a combined phage display peptide library. Antibodies with identical VH and VL or identical CDR1, 2, and 3 sequences are expected to bind to the same epitope.

[0123] An antibody that "competes with another antibody for binding to a target" refers to an antibody that inhibits (partially or completely) another antibody from binding to a target. Whether two antibodies compete with each other for binding to a target, i.e., whether and to what extent an antibody inhibits another antibody from binding to a target, can be determined using known competition experiments. In certain embodiments, an antibody competes with another antibody and inhibits another antibody from binding to a target by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. Depending on which antibody is a "blocking antibody" (i.e., a cold antibody that is first incubated with the target), the level of inhibition or competition can be different. Competition assays can be performed as described, for example, in Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi: 10.1101 / pdb.prot4277 or in Chapter 11 of "Using Antibodies" by Ed Harlow and David Lane, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999. Competing antibodies bind to the same epitope, overlapping epitopes, or adjacent epitopes (e.g., as demonstrated by steric hindrance).

[0124] Other competitive binding assays include: solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)); solid phase direct label assay, solid phase direct label sandwich assay (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid phase direct label RIA using 1-125 label (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid phase direct biotin-avidin EIA (Cheung et al., Virology 176:546 (1990)); and directly labeled RIA (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)).

[0125] "Bispecific" or "bifunctional antibodies" are artificial hybrid antibodies with two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods, including fusion of hybridomas or linking of Fab' fragments. See, for example, Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0126] As used herein, the term "monoclonal antibody" refers to an antibody that displays a single binding specificity and affinity for a specific epitope, or a composition of antibodies in which all antibodies display a single binding specificity and affinity for a specific epitope. Thus, the term "human monoclonal antibody" refers to an antibody or antibody composition that displays a single binding specificity and has variable and optional constant regions derived from human germline immunoglobulin sequences. In some embodiments, human monoclonal antibodies are produced by hybridomas comprising B cells fused to immortalized cells obtained from transgenic non-human animals (e.g., transgenic mice) having a genome comprising human heavy and light chain transgenes.

[0127] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, generated or isolated by recombinant means, such as (a) antibodies isolated from transgenic or transchromosomal animals (e.g., mice) for human immunoglobulin genes or hybridomas prepared therefrom, (b) antibodies isolated from host cells converted to express antibodies, such as antibodies isolated from transfectomas, (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, generated or isolated by any other means involving splicing human immunoglobulin gene sequences to DNA sequences. Such recombinant human antibodies comprise variable and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but include subsequent rearrangements and mutations that occur, for example, during antibody maturation. As known in the art (see, e.g., Lonberg (2005) Nature Biotech. 23(9): 1117-1125), the variable region contains antigen-binding domains encoded by various genes that rearrange to form antibodies specific for foreign antigens. In addition to rearrangement, the variable region can be further modified by multiple single amino acid changes (called somatic mutations or hypermutations) to increase the affinity of the antibody for foreign antigens. The constant region will further change in response to the antigen (i.e., isotype switching). Thus, the rearranged and somatically mutated nucleic acid molecules encoding the light and heavy chain immunoglobulin polypeptides that respond to the antigen cannot have sequence identity with the original nucleic acid molecule, but will be substantially identical or similar (i.e., have at least 80% identity).

[0128] "HuMAb" refers to an antibody with a variable region, wherein the framework region and the CDR region are all derived from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. Antibodies as described herein may include amino acid residues that are not encoded by human germline immunoglobulin sequences (for example, mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which the CDR sequences derived from the germline of another mammalian species such as a mouse have been transplanted onto human framework sequences. The terms "human" antibody and "fully human" antibody are used synonymously.

[0129] "Humanized" antibody refers to an antibody in which some, most or all of the amino acids outside the CDR domains of a non-human antibody are replaced by the corresponding amino acids derived from human immunoglobulin. In some embodiments of the humanized form of an antibody, some, most or all of the amino acids outside the CDR domains have been replaced by amino acids from human immunoglobulin, while some, most or all of the amino acids in one or more CDR regions are unchanged. Amino acid whose small addition, deletion, insertion, substitution or modification is permitted as long as they do not destroy the ability of the antibody to bind to a specific antigen. "Humanized" antibody retains an antigenic specificity similar to the original antibody.

[0130] A "chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0131] As used herein, the term "cross-reactivity" refers to the ability of the antibodies described herein to bind to FAM19A5 from different species. For example, an antibody described herein that binds to human FAM19A5 may also bind to another species of FAM19A5 (e.g., mouse FAM19A5). As used herein, cross-reactivity can be measured by detecting specific reactivity with purified antigens in a binding assay (e.g., SPR, ELISA), or by binding to cells that physiologically express FAM19A5, or otherwise interacting with the cell function. Methods for determining cross-reactivity include standard binding assays as described herein, for example, by using Biacore TM Biacore 2000 SPR instrument (Biacore AB, Uppsala, Sweden) TM Surface plasmon resonance (SPR) analysis, or flow cytometry techniques.

[0132] As used herein, the term "naturally occurring" as applied to an object means that the object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including a virus) is naturally occurring, as is a polypeptide or polynucleotide sequence that can be isolated from a source in nature and has not been intentionally modified by humans in a laboratory.

[0133] "Polypeptide" refers to a chain of at least two consecutively linked amino acid residues, with no upper limit to the length of the chain. One or more amino acid residues in a protein may contain modifications such as, but not limited to, glycosylation, phosphorylation, or disulfide bond formation. A "protein" may comprise one or more polypeptides.

[0134] As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. The nucleic acid molecule can be single-stranded or double-stranded and can be a cDNA.

[0135] As used herein, the term "vector" means a nucleic acid molecule capable of transporting another nucleic acid connected thereto. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop in which an additional DNA segment can be connected. Another type of vector is a viral vector, in which an additional DNA fragment can be connected to a viral genome. Some vectors can replicate autonomously in the host cell in which they are introduced (e.g., bacterial vectors and episomal mammalian vectors with bacterial replication origins). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of the host cell after being introduced into the host cell, and thus replicate together with the host genome. In addition, some vectors can guide the expression of the gene effectively connected thereto. Such vectors are generally referred to as "recombinant expression vectors" (or simply "expression vectors") in this article, and expression vectors useful in recombinant DNA technology are typically in the form of plasmids. In this specification, "plasmid" and "vector" can be used interchangeably because plasmids are the most commonly used vector forms. However, other forms of expression vectors are also included, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses), which perform equivalent functions.

[0136] As used herein, the term "recombinant host cell" (or simply "host cell") means a cell that contains a nucleic acid that is not naturally present in the cell, and may be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms refer not only to the specific subject cell, but also to the progeny of such a cell. Because mutations or environmental influences may cause certain modifications to occur in subsequent generations, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0137] As used herein, the term "linked" refers to the association of two or more molecules. The link can be covalent or non-covalent. The link can also be genetic (i.e., recombinant fusion). Such a link can be achieved using a variety of techniques recognized in the art, such as chemical conjugation and recombinant protein production.

[0138] II. Methods for Treating Neuropathic Pain

[0139] Disclosed herein is a method for treating neuropathic pain in a subject in need thereof, comprising administering to the subject an antagonist (e.g., an isolated monoclonal antibody or its antigen-binding fragment) that specifically binds to FAM19A5. In some embodiments, neuropathic pain is central neuropathic pain, that is, pain caused by damage or impairment (e.g., brain injury and spinal cord injury) affecting any level of the CNS including the central somatosensory nervous system, or pain associated with disease or disorder such as stroke, multiple sclerosis or dorsolateral medullary infarction or the pain caused thereby. In some embodiments, central neuropathic pain can be spontaneous or stimulus-induced. In some embodiments, central neuropathic pain can include dynamic mechanical allodynia and cold allodynia. Symptoms of central neuropathic pain include, for example, common sensations such as burning, tingling, shooting sensation, squeezing sensation, cold pain, paresthesia and dysesthesia (e.g., tingling, numbness, cold and oppression). The distribution range of central neuropathic pain includes from small area to large area, for example in the periorbital area, or covers half the body in the case of stroke or covers the lower body in the case of spinal cord injury, or involves one side of the face and the opposite side of the body or limbs. The central neuropathic pain caused by spinal cord injury includes "plane" pain and "below the plane" pain, "plane" pain is the pain perceived in segmental mode at the injury plane, and "below the plane" pain is the pain felt below the injury plane. In certain embodiments, the method slows down, reverses, alleviates, improves, inhibits or slows down or prevents central neuropathic pain, symptoms associated with pain, potential causes of pain or their combination.

[0140] In certain embodiments, neuropathic pain is peripheral neuropathic pain, a pain caused by damage or impairment (e.g., damage to motor nerves, sensory nerves, autonomic nerves, or a combination thereof) of any level affecting the peripheral nervous system, or pain caused by a disease or disorder or pain associated with a disease or disorder. Damage or impairment of motor nerves is associated with symptoms such as muscle weakness (e.g., muscle weakness in the back, legs, buttocks, or face), painful cramps and fasciculations (uncontrolled muscle twitching visible under the skin), muscle atrophy (severe contraction of muscle size), and reduced reflexes. Damage or impairment of sensory nerves results in a variety of symptoms, including hypersensitivity of pain receptors in the pain and skin, leading to allodynia (e.g., severe pain caused by a stimulus that is usually painless).

[0141] In some embodiments, the method treats one or more types of neuropathic pain, comprising administering a FAM19A5 antagonist (e.g., an anti-FAM19A5 antibody or antigen-binding fragment thereof) to a subject in need thereof. In some embodiments, the neuropathic pain that can be treated with the methods disclosed herein is neuralgia, including but not limited to trigeminal neuralgia (TN) (e.g., pain in the area of ​​the trigeminal nerve in the face or mouth), atypical trigeminal neuralgia (ATN), occipital neuralgia, postherpetic neuralgia (e.g., pain unilaterally distributed in one or more spinal dermatomes or the trigeminal ophthalmic branch), peripheral nerve injury pain (e.g., pain in the innervation area of ​​a diseased nerve, typically outside of trauma, surgery, or compression), glossopharyngeal neuralgia (e.g., irritation of the ninth cranial nerve that causes extreme pain in the throat, tongue, and behind the ear), sciatica, low back pain, and atypical facial pain. In some embodiments, the neuralgia is caused by or is associated with chemical irritation, inflammation, trauma (including surgery), nerve compression, such as by a nearby structure (e.g., a tumor), or infection. In some embodiments, neuropathic pain is afferent blockade pain syndrome, which includes but is not limited to damage to the brain or spinal cord, post-stroke pain, phantom limb pain, paraplegia, brachial plexus avulsion, lumbar radiculopathy. In some embodiments, neuropathic pain is complex regional pain syndrome (CRPS), including CRPS1 and CRPS2. CRPS includes but is not limited to, and in some embodiments, the symptoms associated with CRPS may include severe pain, changes in nails, bones and skin; and increased sensitivity of the affected limb to touch. In some embodiments, neuropathic pain is neuropathy (e.g., central or peripheral). Non-limiting examples of neuropathic pain include, for example, single neuropathic pain (mononeuropathy) and multiple neuropathic pain (polyneuropathy).

[0142] In some embodiments, the neuropathic pain is caused by or is associated with a physical injury, including, for example, (1) traumatic injury or damage, including nerve compression (e.g., nerve crush, nerve stretch, nerve entrapment, or incomplete nerve transection); (2) spinal cord injury (e.g., hemisection of the spinal cord); (3) injury or damage to a peripheral nerve (e.g., motor nerve, sensory nerve, or autonomic nerve, or a combination thereof); (4) amputation; contusion; inflammation (e.g., inflammation of the spinal cord); or surgery; and (5) repetitive stress, including, for example, repetitive, clumsy, and / or forced activities requiring prolonged movement of any joint group (e.g., ulnar neuropathy and carpal tunnel syndrome). In some embodiments, the method treats neuropathic pain caused by or associated with exposure to a toxic agent.

[0143] In some embodiments, the neuropathic pain is caused by or is associated with one or more diseases or disorders including, for example, (1) an ischemic event (e.g., a stroke or heart attack), (2) multiple sclerosis, (3) metabolic and / or endocrine diseases or disorders (e.g., diabetes, metabolic diseases, and acromegaly, a condition caused by overproduction of growth hormone and characterized by abnormal enlargement of skeletal parts, including joints, leading to nerve entrapment and pain), (4) small vessel disease, which results in reduced oxygen supply to peripheral nerves, thereby causing damage to nerve tissue (e.g., vasculitis, i.e., inflammation of blood vessels), (5) autoimmune diseases (e.g., Sjögren's syndrome, lupus, rheumatoid arthritis, and acute inflammatory demyelinating neuropathy, also known as Guillain-Barré syndrome), (6) renal disorders, (7) cancer or tumor (e.g., neoplastic tumors, neuromas, paraneoplastic syndromes, and toxicity from chemotherapeutic agents and radiation in cancer treatment), (8) infection (e.g., infection caused by viruses such as varicella zoster (shingles), Epstein-Barr virus, West Nile virus, cytomegalovirus, and herpes simplex virus, AIDS, or infection caused by bacteria such as Lyme disease, diphtheria, and leprosy), (9) inflammatory disorders, (10) peripheral nerve disorders (e.g., neuromas), (11) hereditary or de novo hereditary disorders (e.g., Charcot-Marie-Tooth disorder), (12) mononeuropathies, (13) polyneuropathies, or combinations thereof. In some embodiments, the neuropathic pain is caused by or associated with diabetes (type I or type II). In some embodiments, the neuropathic pain is diabetic peripheral neuropathy.

[0144] In some embodiments, neuropathic pain is caused by or associated with exposure to infectious agents (including, for example, tick-borne infections, varicella zoster, Epstein-Barr virus, West Nile virus, cytomegalovirus, herpes simplex virus, AIDS), or exposure to toxic agents (e.g., drugs, alcohol, heavy metals (e.g., lead, arsenic, mercury), or exposure to industrial agents (e.g., solvents, fumes from glues), and nitrous oxide).

[0145] In certain embodiments, neuropathic pain is caused by or is associated with physical injury, infection, diabetes, cancer therapy, alcoholism, amputation, multiple sclerosis, herpes zoster, spinal surgery, sciatica (pain along the sciatic nerve), lower back pain, neuralgia such as trigeminal neuralgia (for example, pain in the trigeminal region of the face or mouth), neuropathic pain such as painful polyneuropathy (for example, foot pain, which can extend to include the calf, thigh and hand), or a combination thereof. In certain embodiments, neuropathic pain is trigeminal neuralgia. In certain embodiments, neuropathic pain is associated with muscle weakness in the back, legs, buttocks, or face. In certain embodiments, neuropathic pain is caused by compression of nerves (for example, nerves of the legs, feet, buttocks, or nerves of the facial muscles). In certain embodiments, neuropathic pain includes sciatic nerve injury. In certain embodiments, neuropathic pain is sciatica.

[0146] In certain embodiments, the method of the present disclosure can reverse, alleviate, improve, suppress, slow down or prevent one or more symptoms associated with neuropathic pain. Therefore, in one aspect, the present disclosure provides a method for improving the hyperalgesia of a subject in need thereof, including administering an antagonist for FAM19A5 to the subject. As used herein, the term "hyperalgesia" refers to an increased or aggravated reaction to painful stimuli (e.g., acupuncture or hot plate). In certain embodiments, hyperalgesia is directed to mechanical stimuli, such as acupuncture (mechanical hyperalgesia). In other embodiments, hyperalgesia is directed to thermal stimuli, such as hot plate (thermal hyperalgesia). In certain embodiments, a subject in need thereof suffers from chronic contractile injury (e.g., sciatica). In certain embodiments, a subject in need thereof suffers from diabetic peripheral neuropathy.

[0147] In some embodiments, administering a FAM19A5 antagonist to a subject in need thereof allows the subject to have a higher threshold to mechanical stimulation compared to a reference control (e.g., a neuropathic pain subject who has not received a FAM19A5 antagonist). As used herein, the term "threshold to mechanical stimulation" refers to the amount of pressure (from a mechanical stimulation) before the subject reacts to the stimulation (e.g., by pulling apart). Thus, a subject with a higher threshold can withstand or resist a greater amount of mechanical stimulation than a subject with a lower threshold. In some embodiments, the methods disclosed herein can increase a subject's threshold to mechanical stimulation by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, or at least 200%, compared to a reference control (e.g., a subject's threshold before administering a FAM19A5 antagonist).

[0148] In some embodiments, administration of a FAM19A5 antagonist to a subject in need thereof increases the subject's latency to a thermal stimulus (e.g., a hot plate) (i.e., the time interval between the stimulus and the response) compared to a reference control (e.g., a subject with neuropathic pain who did not receive the FAM19A5 antagonist). In some embodiments, the methods disclosed herein can increase the subject's latency to a thermal stimulus by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, or at least 200% compared to a reference control (e.g., the subject's threshold before administration of the FAM19A5 antagonist).

[0149] In another aspect, the present disclosure provides a method for improving sensory nerve conduction velocity in a subject in need thereof. As used herein, the term "sensory nerve conduction velocity" (SNCV) refers to the rate at which electrical signals travel through peripheral nerves. Healthy nerves send electrical signals faster and with greater intensity than damaged nerves. See Chouhan S., J Clin Diagn Res 10(1):CC01-3(2016). Therefore, tests that help measure SNCV (e.g., sensory nerve conduction velocity tests) can be used to identify potential nerve damage and / or dysfunction in a subject. In some embodiments, the methods disclosed herein can increase the SNCV of a subject with neuropathic pain by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, or at least 200%, compared to a reference control (e.g., a subject's threshold value before administration of a FAM19A5 antagonist).

[0150] There are animal models for studying neuropathic pain. Non-limiting examples of such animal models include (1) the spinal nerve ligation (SNL) model, in which one or more spinal nerves leading to the foot are ligated and severed (see Kim SH and Chung JM., Pain 50:355–363 (1992)); (2) the partial sciatic nerve ligation (PSL) model, in which a portion of the sciatic nerve is tightly ligated (see Seltzer et al., Pain 43:205–218 (1990)); (3) the chronic constriction injury (CCI) model, which involves placing four loose chromic gut ligatures on the sciatic nerve, and an immune response to the sutures results in nerve swelling and nerve constriction (see, e.g., Example 6); (4) the sparing nerve injury (SNI) model, in which the common peroneal and tibial nerves are severed, sparing the sural nerve (see Decosterd I and Woolf CJ, Pain 87:149-158 (2000)); and (5) streptozotocin (STZ)-induced diabetic rats, in which injection of STZ causes pancreatic swelling and degeneration of β cells of the islets of Langerhans, thereby causing experimental diabetes in rats. (See, e.g., Example 8; Akbarzadeh A et al., Indian J. Clin. Biochem. 22(2):60-64 (2007)). These models result in hyperalgesia in animals, manifested as increased responses to mechanical and / or thermal stimuli.

[0151] Tests for mechanical hyperalgesia in animals include the von Frey test, in which von Frey monofilaments with varying bending forces are applied to the plantar surface of the foot. Following nerve injury, the threshold for paw withdrawal is significantly reduced. Li et al., Pain 85:493–502 (2000); see also Example 6. Thus, in some embodiments, the methods disclosed herein can increase the threshold for paw withdrawal in animal models of neuropathic pain (e.g., chronic constriction injury models).

[0152] Testing for thermal (heat) hyperalgesia can include using a radiant heat source (e.g., a hot plate) focused on the plantar surface of the foot and measuring the reaction time to paw withdrawal. After nerve injury, the foot withdraws faster than before the injury. Kim SH and Chung JM. Pain 50:355-363 (1992); see also Example 8. In some embodiments, the methods disclosed herein can increase the latency of paw withdrawal in an animal model of neuropathic pain (e.g., a diabetic peripheral neuropathy model).

[0153] In some embodiments, the subject treated with the methods of the present disclosure is a non-human animal, such as a rat or mouse. In some embodiments, the subject treated in the methods is a human.

[0154] In some embodiments, a FAM19A5 antagonist (e.g., an anti-FAM19A5 antibody or antigen-binding portion thereof), bispecific molecule, or immunoconjugate disclosed herein, or a composition thereof, is administered intravenously, orally, parenterally, intrathecally, intracerebroventricularly, intrapulmonary, intramuscularly, subcutaneously, intraperitoneally, intravitreally, or intracerebroventricularly.

[0155] In some embodiments, a FAM19A5 antagonist or composition thereof can be administered in combination with one or more additional agents for treating neuropathic pain. For example, non-limiting exemplary agents for treating neuropathic pain include venlafaxine ( ), antiepileptic drugs such as carbamazepine ( , FDA-approved for the relief of pain from trigeminal neuralgia), gabapentin ( , approved to treat postherpetic neuralgia (PHN): pain that lasts one to three months after shingles heals) and pregabalin ( , approved for PHN, painful diabetic neuropathic pain, and fibromyalgia), sodium channel blockers such as lidocaine, adrenergic drugs such as clonidine, phentolamine, phenoxybenzamine, reserpine, dexmedetomidine, opioids such as morphine, and antidepressants such as amitriptyline, imipramine, and duloxetine.

[0156] Dosages and administration of one or more additional therapeutic agents are known in the art, for example, as indicated by the product labeling of the corresponding agent.

[0157] III. FAM19A5 antagonists

[0158] One or more FAM19A5 antagonists can be used to treat neuropathic pain. In some embodiments, the FAM19A5 antagonist is an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA (peptide nucleic acid) or a vector comprising the same that specifically targets FAM19A5. In other embodiments, the FAM19A5 antagonist is an antibody or antigen-binding fragment thereof that specifically binds to the FAM19A5 protein, a polynucleotide encoding an anti-FAM19A5 antibody or an antigen-binding fragment thereof, or a vector comprising the polynucleotide thereof.

[0159] Antibodies useful in the methods disclosed herein include monoclonal antibodies characterized by specific functional characteristics or properties. For example, the antibodies specifically bind to human FAM19A5, including soluble FAM19A5 and membrane-bound FAM19A5. In addition to specifically binding to soluble and / or membrane-bound human FAM19A5, the antibodies described herein also (a) bind to soluble human FAM 19A5 with a KD of 10 nM or less; (b) bind to membrane-bound human FAM19A5 with a KD of 10 nM or less; or both (a) and (b).

[0160] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to the FAM19A5 antigen with high affinity (e.g., 10-7 M or less, 10-8 M or less, 10-9 M (1 nM) or less, 10-10 M (0.1 nM) or less, 10-11 M or less, or 10-12 M or less, e.g., 10-12 M to 10-7 M, 10-11 M to 10-7 M, 10-10 M or less). To 10-7M, or 10-9M to 10-7M, for example, a KD of 10-12M, 5×10-12M, 10-11M, 5×10-11M, 10-10M, 5×10-10M, 10-9M, 5×10-9M, 10-8M, 5×10-8M, 10-7M or 5×10-7M) specifically binds to soluble human FAM19A5 or membrane-bound human. Standard assays for assessing the binding ability of antibodies to human FAM19A5 of different species are known in the art, including, for example, ELISA, Western blot and RIA. Suitable assays are described in detail in the Examples. The binding kinetics (e.g., binding affinity) of the antibody can also be assessed by standard assays known in the art, such as by ELISA, BIACORE™ analysis or Assays for evaluating the effects of antibodies on the functional properties of FAM19A5 (eg, ligand binding) are described in further detail below and in the Examples.

[0161] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to soluble human FAM19A5 with a KD of 10-7 M or less, 10-8 M (10 nM) or less, 10-9 M (1 nM) or less, 10-10 M or less, 10-12 M to 10-7 M, 10-11 M to 10-7 M, 10-10 M to 10-7 M, 10-9 M to 10-7 M, or 10-8 M to 10-7 M, e.g., as determined by ELISA. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to soluble FAM19A5 with a KD of 10 nM or less, e.g., between 0.1 and 10 nM, between 0.1 and 5 nM, between 0.1 and 1 nM, between 0.5 and 10 nM, between 0.5 and 5 nM, between 0.5 and 1 nM, between 1 and 10 nM, between 1 and 5 nM, or between 5 and 10 nM. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, is expressed at about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 200 pM, 300 pM, 400 pM, The invention further comprises a novel oligonucleotide that specifically binds to soluble human FAM19A5 with a KD of pM, 500 pM, 600 pM, 700 pM, 800 pM, or 900 pM, or about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, or 9 nM, or about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM.

[0162] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to membrane-bound human with a KD of 10-7 M or less, 10-8 M (10 nM) or less, 10-9 M (1 nM) or less, 10-10 M or less, 10-12 M to 10-7 M, 10-11 M to 10-7 M, 10-10 M to 10-7 M, 10-9 M to 10-7 M, or 10-8 M to 10-7 M, e.g., as determined by ELISA. In certain embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, specifically binds to membrane-bound human FAM19A5 with a KD of 10 nM or less, e.g., between 0.1 and 10 nM, between 0.1 and 5 nM, between 0.1 and 1 nM, between 0.5 and 10 nM, between 0.5 and 5 nM, between 0.5 and 1 nM, between 1 and 10 nM, between 1 and 5 nM, or between 5 and 10 nM, as determined by ELISA. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, is expressed at about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 200 pM, 300 pM, 400 pM, The IL-17 antibodies bind to membrane-bound human FAM19A5 with a K of 100 pM, 500 pM, 600 pM, 700 pM, 800 pM, or 900 pM, or about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, or 9 nM, or about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM.

[0163] In some embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, suitable for the methods disclosed herein cross-competes for binding to (or inhibits binding to) a human FAM19A5 epitope with an anti-FAM19A5 antibody comprising a CDR or variable region disclosed herein.

[0164] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, inhibits binding of a reference antibody comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, (i) wherein the heavy chain CDR1, CDR2, and CDR3 of the reference antibody comprise the amino acid sequences of SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, respectively, and the light chain CDR1, CDR2, and CDR3 of the reference antibody comprise the amino acid sequences of SEQ ID NO: 23, SEQ ID NO: 24, and SEQ ID NO: 25, respectively; (ii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 15, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 27, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: NO:28; (iii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:17, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:18, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:19, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:29, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:30, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:31; (iv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:20, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:21, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:22, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:32, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:33, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:34; (v) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:89, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: NO:90, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:91, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:92, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:93, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:94;(vi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 95, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 96, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 97, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 98, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 99, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 100; (vii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 101, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 102, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 103, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 104, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 105, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 106; (viii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 107, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 108 NO: 108, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 109, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 110, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 111, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 112; (ix) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 113, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 114, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 115, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 116, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 117, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 118; (x) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 119, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 120, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 121. NO: 121, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 122, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 123, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 124;(xi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 125, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 126, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 127, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 128, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 129, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 130; (xii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 131, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 132, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 133, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 134, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 135, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 136; (xiii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 137, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 138 NO: 138, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 139, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 140, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 141, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 142; (xiv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 143, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 144, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 145, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 146, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 147, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 148; or (xv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 149, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 150, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 151 NO: 151, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 152, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 153, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 154. In some embodiments, the heavy chain CDR1, CDR2, and CDR3 of the reference antibody comprise the CDR1, CDR2, and CDR3 sequences, respectively, as set forth in Table 2, and the light chain CDR1, CDR2, and CDR3 of the reference antibody comprise the CDR1, CDR2, and CDR3 sequences, respectively, as set forth in Table 3.

[0165] In some embodiments, the reference antibody comprises (a) heavy and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively; (b) heavy and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively; (c) heavy and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively; (d) heavy and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively; (e) heavy and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively; (f) heavy and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively; (g) heavy and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively; (h) heavy and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively; (i) heavy and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively; (j) heavy and light chain variable region sequences comprising SEQ ID NOs: 171 and 172, respectively. (k) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 161 and 172, respectively; (l) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 162 and 173, respectively; (m) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 163 and 174, respectively; (n) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 164 and 175, respectively; or (o) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 165 and 176, respectively. In some embodiments, the reference antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence as set forth in Table 4, and the VL comprises an amino acid sequence as set forth in Table 5.

[0166] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, inhibits the binding of such a reference antibody to human FAM19A5 by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. Competing antibodies bind to the same epitope, overlapping epitopes, or adjacent epitopes (e.g., as demonstrated by steric hindrance). Competition assays known in the art, such as RIAs and EIAs, can be used to determine whether two antibodies compete with each other for binding to a target.

[0167] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to the same FAM19A5 epitope as a reference antibody disclosed herein, the reference antibody comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 11, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 12, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 13, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 23, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 25; or (ii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 15, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: NO:27, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:28; (iii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:17, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:18, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:19, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:29, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:30, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:31; (iv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:20, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:21, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:22, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:32, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:33, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:34; (v) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: NO:89, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:90, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:91, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:92, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:93, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:94;(vi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 95, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 96, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 97, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 98, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 99, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 100; (vii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 101, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 102, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 103, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 104, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 105, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 106; (viii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 107, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 108 NO: 108, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 109, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 110, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 111, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 112; (ix) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 113, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 114, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 115, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 116, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 117, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 118; (x) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 119, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 120, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 121. NO: 121, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 122, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 123, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 124;(xi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 125, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 126, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 127, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 128, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 129, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 130; (xii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 131, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 132, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 133, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 134, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 135, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 136; (xiii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 137, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 138 NO: 138, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 139, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 140, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 141, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 142; (xiv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 143, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 144, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 145, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 146, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 147, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 148; or (xv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 149, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 150, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 151 The light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 152, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 153, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 154.

[0168] In some embodiments, the reference antibody comprises (a) heavy and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively; (b) heavy and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively; (c) heavy and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively; (d) heavy and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively; (e) heavy and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively; (f) heavy and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively; (g) heavy and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively; (h) heavy and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively; (i) heavy and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively; (j) heavy and light chain variable region sequences comprising SEQ ID NOs: 171 and 172, respectively. NOs:160 and 171; (k) heavy chain and light chain variable region sequences comprising SEQ ID NOs:161 and 172, respectively; (l) heavy chain and light chain variable region sequences comprising SEQ ID NOs:162 and 173, respectively; (m) heavy chain and light chain variable region sequences comprising SEQ ID NOs:163 and 174, respectively; (n) heavy chain and light chain variable region sequences comprising SEQ ID NOs:164 and 175, respectively; or (o) heavy chain and light chain variable region sequences comprising SEQ ID NOs:165 and 176, respectively.

[0169] Techniques for determining whether two antibodies bind to the same epitope include, for example, epitope mapping methods such as X-ray analysis of crystals of the antigen:antibody complex (which provides atomic resolution of the epitope) and hydrogen / deuterium exchange mass spectrometry (HDX-MS), methods for monitoring antibody binding to antigen fragments or mutated variants of the antigen, where loss of binding due to modification of amino acid residues in the antigen sequence is generally considered indicative of epitope composition, and computational combinatorial methods for epitope mapping.

[0170] The anti-FAM19A5 antibodies or antigen-binding portions thereof useful in the methods disclosed herein can bind to at least one epitope of mature human FAM19A5, as determined by, for example, binding of the antibody to a fragment of human FAM19A5. In some embodiments, the anti-FAM19A5 antibodies or antigen-binding fragments thereof bind to a fragment located within the amino acid sequence of TLDRDSSQPRRTIARQTARC (SEQ ID NO: 6 or amino acid residues 42 to 61 of SEQ ID NO: 2), such as an epitope having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids of SEQ ID NO: 6. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to SEQ ID NO: 6 at one or more amino acids corresponding to amino acid residues 46 to 51 (i.e., DSSQPR), e.g., amino acid residues 46, 50, and 52 (i.e., D---PR), e.g., amino acid residues 46, 47, 48, and 50 (i.e., DSS-P) of SEQ ID NO: 2. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding fragment thereof, binds to an epitope located within the amino acid sequence of CDMLPCLEGEGCDLLINRSG (SEQ ID NO: 9 or amino acids 90 to 109 of SEQ ID NO: 2), e.g., an epitope having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids of SEQ ID NO: 9. In certain embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to SEQ ID NO: 9 at one or more amino acid residues 99 to 107 of SEQ ID NO: 4 (i.e., EGCDLLINR), e.g., amino acid residues 102, 103, 105, and 107 (i.e., DL-IR), e.g., amino acid residues 99, 100, 102, 103, 105, and 107 (i.e., EG-DL-IR), e.g., amino acid residues 99, 100, and 107 (i.e., EG------R).

[0171] In some embodiments, at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 6. In some embodiments, at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 9.

[0172] In some embodiments, the anti-FAM19A5 antibody, or antigen binding portion thereof, binds only to a human FAM19A5 epitope that is SEQ ID NO: 5, 6, 7, 8, 9, or 10, or a fragment within the amino acid sequence of SEQ ID NO: 5, 6, 7, 8, 9, or 10, such as an epitope having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids of SEQ ID NO: 5, 6, 7, 8, 9, or 10.

[0173] In some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof of the present disclosure bind to SEQ ID NO: 6 or a fragment thereof in its native configuration (i.e., undenatured). In some embodiments, the anti-FAM19A5 antibodies or antigen-binding fragments thereof of the present disclosure bind to SEQ ID NO: 9 or a fragment thereof in its native configuration (i.e., undenatured). In some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof of the present disclosure bind to glycosylated and unglycosylated human FAM19A5.

[0174] In some embodiments, the anti-FAM19A5 antibody or antigen-binding portion thereof further binds to one or more additional FAM19A5 epitopes. Thus, certain anti-FAM19A5 antibodies or antigen-binding portions thereof bind to (i) the epitope of SEQ ID NO: 6 and an additional epitope, or (ii) the epitope of SEQ ID NO: 9 and an additional epitope. Other anti-FAM19A5 antibodies or antigen-binding fragments thereof may bind to the epitope of SEQ ID NO: 5, SEQ ID NO: 9, and an additional epitope. In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof binds to the epitope of SEQ ID NO: 6, the epitope of SEQ ID NO: 10, and an additional epitope.

[0175] In some embodiments, one or more additional FAM19A5 epitopes are selected from QLAAGTCEIVTLDR (SEQ ID NO:5, epitope F1), TLDRDSSQPRRTIARQTARC (SEQ ID NO:6, epitope F2), TARCACRKGQIAGTTRARPA (SEQ ID NO:7, epitope F3), ARPACVDARIIKTKQWCDML (SEQ ID NO:8, epitope F4), CDMLPCLEGEGCDLLINRSG (SEQ ID NO:9, epitope F5) or NRSGWTCTQPGGRIKTTTVS (SEQ ID NO:10, epitope F6) or a fragment located within the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, or any combination thereof. Fragments located within the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10 include fragments having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of any one of SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10. In some embodiments, the one or more additional FAM19A5 epitopes are selected from SEQ ID NO: 5, 6, 7, 8, 9 or 10, or fragments located within the amino acid sequence of SEQ ID NO: 5, 6, 7, 8, 9 or 10, such as fragments having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of SEQ ID NO: 5, 6, 7, 8, 9 or 10, or any combination thereof. In some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof of the present disclosure bind to any of the one or more additional epitopes in their native configuration (i.e., not denatured). In some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof bind to one or more additional glycosylated and non-glycosylated FAM19A5 epitopes.

[0176] In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to at least one FAM19A5 epitope identified as EP2, EP4, and / or EP8, wherein EP2 comprises, consists essentially of, or consists of the amino acids DSSQP (SEQ ID NO: 66), wherein EP4 comprises, consists essentially of, or consists of the amino acids ARCACRK (SEQ ID NO: 68), and wherein EP8 comprises, consists essentially of, or consists of the amino acids TCTQPGGR (SEQ ID NO: 72). In some embodiments, at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to EP2, EP4, or EP8. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds only to EP2. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to both EP4 and EP8.

[0177] In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to at least one FAM19A5 epitope identified as EP6, EP7, or EP8, wherein EP6 comprises the amino acids KTKQWCDML (SEQ ID NO: 70), wherein EP7 comprises the amino acids GCDLLINR (SEQ ID NO: 71), and wherein EP8 comprises the amino acids TCTQPGGR (SEQ ID NO: 72). In some embodiments, at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to EP6, EP7, or EP8. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds only to EP6, EP7, or EP8. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to EP6, EP7, and EP8. In some embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to EP7 and EP8. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to EP7.

[0178] In some embodiments, the anti-FAM19A5 antibody, or antigen binding portion thereof, binds to one or more FAM19A5 epitopes selected from the group consisting of SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, and any combination thereof.

[0179] In some embodiments, provided herein are antibodies, or antigen-binding portions thereof, that bind to FAM19A5 (e.g., human FAM19A5) with 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more affinity compared to another protein in the FAM19A family, as measured by, for example, an immunoassay (e.g., ELISA), surface plasmon resonance, or kinetic exclusion assay. In specific embodiments, provided herein are antibodies, or antigen-binding portions thereof, that bind to FAM19A5 (e.g., human FAM19A5) that have no cross-reactivity with another protein in the FAM19A family, as measured by, for example, an immunoassay.

[0180] In some embodiments, the anti-FAM19A5 antibody is not a natural antibody or is not a naturally occurring antibody. For example, the anti-FAM19A5 antibody has different post-translational modifications than naturally occurring antibodies, such as having more, fewer, or different types of post-translational modifications.

[0181] IV. Exemplary Anti-FAM19A5 Antibodies

[0182] Specific antibodies that can be used in the methods disclosed herein are antibodies (e.g., monoclonal antibodies) having the CDR and / or variable region sequences of antibodies 1-65 isolated in Example 1, and antibodies having at least 80% identity (e.g., at least 85%, at least 90%, at least 95%, or at least 99% identity) to their variable region or CDR sequences. The amino acid sequences of the VH and VL CDRs of different anti-FAM19A5 antibodies are provided in Tables 2 and 3, respectively. The CDRs of the following antibodies were identified using the Kabat numbering scheme (see above): 1-65, 3-2, 2-13, 1-28, P2-C12, 13B4, 13F7, 15A9, P1-A03, P1-A08, P1-F02, P2-A01, P2-A03, P2-F07, P2-F11, SS01-13, SS01-13-s5, and S5-2.GKNG. The CDRs of the following antibodies were identified using the IMGT numbering system (see above): 1-7A-IT, low-PI, 1-30, 1-17, 1-32, 4-11, 6-10, 2-13D, 2-13D-37, 2-13D-37-1.5W-41, and 2-13D-37-3W-16. The VH and VL amino acid sequences of various anti-FAM19A5 antibodies of the present disclosure are provided in Tables 4 and 5, respectively.

[0183] Table 2 Variable heavy chain CDR amino acid sequences

[0184]

[0185]

[0186]

[0187] Table 3 Variable light chain CDR amino acid sequences

[0188]

[0189]

[0190]

[0191] Table 4: Variable heavy chain amino acid sequences

[0192]

[0193]

[0194]

[0195] Table 5: Variable light chain amino acid sequences

[0196]

[0197]

[0198] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271, or 275. In other embodiments, the isolated anti-FAM19A5 antibody or antigen-binding portion thereof comprises a CDR of the heavy chain variable region selected from the group consisting of SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271, or 275.

[0199] In some embodiments, the isolated anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NOs: 39-42, 166-176, 204, 227-234, or 272. In other embodiments, the isolated anti-FAM19A5 antibody or antigen-binding portion thereof comprises a light chain variable region CDR selected from the group consisting of SEQ ID NOs: 39-42, 166-176, 227-234, or 272.

[0200] In some embodiments, the isolated anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises a heavy chain variable region CDR selected from the group consisting of SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271, or 275 and a light chain variable region CDR selected from the group consisting of SEQ ID NOs: 39-42, 166-176, 227-234, or 272.

[0201] In some embodiments, the isolated anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, (i) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 37 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 41; (ii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 36 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 40; (iii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 35 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 39; (iv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 38 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 42; (v) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 155 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 166; (vi) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 156 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: NO: 167; (vii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 157, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 168; (viii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 158, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 169; (ix) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 159, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 170; (x) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 160, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 171; (xi) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 161, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 172; (xii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 162, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: NO:173; (xiii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:163, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:174; (xiv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:164, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:175; and (xv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:165, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:176.In some embodiments, the isolated anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein VH comprises a VH sequence as set forth in Table 4, and VL comprises a VL sequence as set forth in Table 5.

[0202] In some embodiments, the isolated anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271, or 275.

[0203] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NOs: 39-42, 166-176, 227-234, or 272.

[0204] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271, or 275, and wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NOs: 39-42, 166-176, 227-234, or 272.

[0205] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0206] (a) heavy and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively;

[0207] (b) heavy and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively;

[0208] (c) heavy and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively;

[0209] (d) heavy and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively;

[0210] (e) heavy and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively;

[0211] (f) heavy and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively;

[0212] (g) heavy and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively;

[0213] (h) heavy and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively;

[0214] (i) heavy and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively;

[0215] (j) heavy and light chain variable region sequences comprising SEQ ID NOs: 160 and 171, respectively;

[0216] (k) heavy and light chain variable region sequences comprising SEQ ID NOs: 161 and 172, respectively;

[0217] (1) heavy and light chain variable region sequences comprising SEQ ID NOs: 162 and 173, respectively;

[0218] (m) heavy and light chain variable region sequences comprising SEQ ID NOs: 163 and 174, respectively;

[0219] (n) heavy and light chain variable region sequences comprising SEQ ID NOs: 164 and 175, respectively; or

[0220] (o) heavy and light chain variable region sequences comprising SEQ ID NOs: 165 and 176, respectively.

[0221] In certain embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, comprises (i) a heavy chain CDR1, CDR2, and CDR3 of 1-65, or a combination thereof, and / or a light chain CDR1, CDR2, and CDR3 of 1-65, or a combination thereof; (ii) a heavy chain CDR1, CDR2, and CDR3 of 3-2, or a combination thereof, and / or a light chain CDR1, CDR2, and CDR3 of 3-2, or any combination thereof; (iii) a heavy chain CDR1, CDR2, and CDR3 of 2-13, or a combination thereof, and / or a light chain CDR1, CDR2, and CDR3 of 2-13, or any combination thereof; (iv) a heavy chain CDR1, CDR2, and CDR3 of 1-28, or a combination thereof. R3 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 1-28 or any combination thereof; (v) the heavy chain CDR1, CDR2 and CDR3 of P2-C12 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of P2-C12 or any combination thereof; (vi) the heavy chain CDR1, CDR2 and CDR3 of 13B4 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 13B4 or any combination thereof; (vii) the heavy chain CDR1, CDR2 and CDR3 of 13F7 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 13F7 or any combination thereof; viii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 15A9, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 15A9; (ix) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A03, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A03; (x) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A08, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A08; (xi) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P1-F02, and / or P1 -F02 or any combination thereof; (xii) the heavy chain CDR1, CDR2 and CDR3 of P2-A01 or any combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of P2-A01 or any combination thereof; (xiii) the heavy chain CDR1, CDR2 and CDR3 of P2-A03 or any combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of P2-A03 or any combination thereof; (xiv) the heavy chain CDR1, CDR2 and CDR3 of P2-F07 or any combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of P2-F07 or any combination thereof;or (xv) the heavy chain CDR1, CDR2, and CDR3 of P2-F11, or any combination thereof, and / or the light chain CDR1, CDR2, and CDR3 of F2-F11, or any combination thereof. The amino acid sequences of the VH CDR1, CDR2, and CDR3 of various anti-FAM19A5 antibodies disclosed herein are provided in Table 2. The amino acid sequences of the VL CDR1, CDR2, and CDR3 of various anti-FAM19A5 antibodies disclosed herein are provided in Table 3.

[0222] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0223] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 17; and / or

[0224] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 18; and / or

[0225] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO: 19.

[0226] In some embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VH CDRs.

[0227] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0228] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 29; and / or

[0229] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 30; and / or

[0230] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 31.

[0231] In some embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VL CDRs.

[0232] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0233] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 17;

[0234] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 18;

[0235] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 19;

[0236] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 29;

[0237] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 30; and / or

[0238] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 31.

[0239] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0240] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 14;

[0241] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15; and / or

[0242] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO: 16.

[0243] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VH CDRs.

[0244] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0245] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 26;

[0246] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27; and / or

[0247] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 28.

[0248] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VL CDRs.

[0249] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0250] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 14;

[0251] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15;

[0252] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 16;

[0253] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 26;

[0254] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27; and / or

[0255] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 28.

[0256] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof that specifically binds to human FAM19A5 comprises:

[0257] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 11;

[0258] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 12; and / or

[0259] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO: 13.

[0260] In some embodiments, the antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VH CDRs.

[0261] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof that specifically binds to human FAM19A5 comprises:

[0262] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 23;

[0263] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 24; and / or

[0264] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 25.

[0265] In some embodiments, the antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VL CDRs.

[0266] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof that specifically binds to human FAM19A5 comprises:

[0267] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 11;

[0268] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 12;

[0269] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 13;

[0270] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 23;

[0271] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 24; and / or

[0272] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 25.

[0273] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0274] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 20;

[0275] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 21; and / or

[0276] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0277] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VH CDRs.

[0278] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0279] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 32;

[0280] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and / or

[0281] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 34.

[0282] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VL CDRs.

[0283] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0284] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 20;

[0285] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 21;

[0286] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 22;

[0287] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 32;

[0288] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and / or

[0289] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO: 34.

[0290] In some embodiments, an anti-FAM19A5 antibody disclosed herein, or an antigen-binding portion thereof, comprises one, two, three, four, five, or six of the above-described CDRs.

[0291] The VH domains described herein, or one or more CDRs thereof, can be linked to constant domains to form heavy chains, such as full-length heavy chains. Similarly, the VL domains described herein, or one or more CDRs thereof, can be linked to constant domains to form light chains, such as full-length light chains. The full-length heavy chains and full-length light chains combine to form a full-length antibody.

[0292] Therefore, in a specific embodiment, an antibody comprising an antibody light chain and a heavy chain (e.g., a separate light chain and a heavy chain) that can be used for the methods disclosed herein is provided. With respect to the light chain, in a specific embodiment, the light chain of the antibody described herein is a kappa light chain. In another specific embodiment, the light chain of the antibody described herein is a lambda light chain. In yet another specific embodiment, the light chain of the antibody described herein is a human kappa light chain or a human lambda light chain. In a specific embodiment, the antibody specifically binding to the FAM19A5 polypeptide (e.g., human FAM19A5) described in the methods disclosed herein comprises a light chain comprising any VL or VL CDR amino acid sequence described herein, and wherein the constant region of the light chain comprises the amino acid sequence of a human kappa light chain constant region. In a specific embodiment, the antibody specifically binding to the FAM19A5 polypeptide (e.g., human FAM19A5) described in the methods disclosed herein comprises a light chain comprising a VL or VL CDR amino acid sequence described herein, and wherein the constant region of the light chain comprises the amino acid sequence of a human lambda light chain constant region. Non-limiting examples of human constant region sequences have been described in the art; see, eg, US Pat. No. 5,693,780 and Kabat EA et al. (1991) supra.

[0293] With regard to the heavy chain, in some embodiments, the heavy chain of the antibodies described herein may be an alpha (α), delta (δ), epsilon (ε), gamma (γ) or mu (μ) heavy chain. In another specific embodiment, the heavy chain of the antibodies described may comprise a human alpha (α), delta (δ), epsilon (ε), gamma (γ) or mu (μ) heavy chain. In some embodiments, the antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) that can be used in the methods disclosed herein comprise a heavy chain comprising a VH or VH CDR amino acid sequence described herein, and wherein the constant region of the heavy chain comprises the amino acid sequence of a human gamma (γ) heavy chain constant region. In other embodiments, the antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) comprise a heavy chain comprising a VH or VH CDR amino acid sequence disclosed herein, and wherein the constant region of the heavy chain comprises the amino acid sequence of a human heavy chain described herein or known in the art. Non-limiting examples of human constant region sequences have been described in the art; see, eg, US Pat. No. 5,693,780 and Kabat EA et al., (1991) supra.

[0294] In some embodiments, the antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) that can be used in the methods disclosed herein comprise a VL domain and a VH domain comprising a VH or VH CDR and a VL and VL CDR as described herein, and wherein the constant region comprises an amino acid sequence of a constant region of an IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule or a human IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule. In another specific embodiment, the antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) comprise a VL domain and a VH domain comprising any amino acid sequence described herein, and wherein the constant region comprises an amino acid sequence of a constant region of an IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule, any subclass of immunoglobulin molecules (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2). In some embodiments, the constant region comprises the amino acid sequence of the constant region of naturally occurring human IgG, including subclasses (e.g., IgG1, IgG2, IgG3, or IgG4) and allotypes (e.g., G1m, G2m, G3m, and nG4m) and variants thereof. See, e.g., Vidarsson G et al. Front Immunol. 5:520 (published online on October 20, 2014) and Jefferis R and Lefranc MP, mAbs 1:4, 1-7 (2009). In some embodiments, the constant region comprises the amino acid sequence of the constant region of human IgG1, IgG2, IgG3, or IgG4, or variants thereof.

[0295] In certain embodiments, the disclosed anti-FAM19A5 antibodies or antigen-binding portions thereof that can be used in the methods disclosed herein do not have Fc effector functions, such as complement-dependent cytotoxicity (CDC) and / or antibody-dependent cellular phagocytosis (ADCP). Effector functions are mediated by the Fc region, and the residues closest to the hinge region in the CH2 domain of the Fc region are responsible for the effector functions of the antibody because it contains largely overlapping binding sites for C1q (complement) and IgG-Fc receptors (FcγRs) on effector cells of the innate immune system. Moreover, IgG2 and IgG4 antibodies have lower levels of Fc effector functions than IgG1 and IgG3 antibodies. Effector functions of antibodies can be reduced or avoided by various methods known in the art, including (1) using antibody fragments lacking an Fc region (e.g., such as Fab, F(ab')2, single-chain Fv (scFv), or sdAbs composed of monomeric VH or VL domains); (2) producing aglycosylated antibodies, which can be produced by, for example, deleting or changing the residue to which the sugar is attached, enzymatically removing the sugar, producing the antibody in cells cultured in the presence of a glycosylation inhibitor, or by expressing the antibody in cells that cannot glycosylate proteins (e.g., bacterial host cells, see, e.g., U.S. Patent Publication No. 20120100140); (3) employing an Fc region from an IgG subclass with reduced effector function (e.g., an Fc region from an IgG2 or IgG4 antibody or a chimeric Fc region comprising a CH2 domain from an IgG2 or IgG4 antibody, see, e.g., U.S. Patent Publication No. 20120100140 and Lau et al.). C et al. J. Immunol. 191:4769-4777 (2013); and (4) generating Fc regions with mutations that result in reduced or no Fc function. See, e.g., U.S. Patent Publication No. 20120100140 and the U.S. and PCT applications cited therein and An et al. mAbs 1:6, 572-579 (2009).

[0296] Thus, in some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof useful in the methods disclosed herein are Fab, Fab', F(ab')2, Fv, single-chain Fv (scFv), or sdAbs consisting of monomeric VH or VL domains. Such antibody fragments are well known in the art and are described above.

[0297] In some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof useful in the methods disclosed herein comprise an Fc region with reduced or no Fc effector function. In some embodiments, the constant region comprises the amino acid sequence of the Fc region of human IgG2 or IgG4, and in some embodiments, the anti-FAM19A5 antibody is of the IgG2 / IgG4 isotype. In some embodiments, the anti-FAM19A5 antibody comprises: a chimeric Fc region comprising a CH2 domain from an IgG antibody of the IgG4 isotype and a CH3 domain from an IgG antibody of the IgG1 isotype; or a chimeric Fc region comprising a hinge region from IgG2 and a CH2 region from IgG4; or an Fc region having a mutation that results in reduced or no Fc function. Fc regions with reduced or no Fc effector function include those known in the art. See, e.g., Lau C. et al., J. Immunol. 191: 4769-4777 (2013); An et al., mAbs 1: 6, 572-579 (2009); and U.S. Patent Publication No. 20120100140 and the U.S. patents and publications and PCT publications cited therein. One of ordinary skill in the art can also readily prepare an Fc region with reduced or no Fc effector function.

[0298] V. Nucleic Acid Molecules

[0299] Another aspect described herein relates to one or more nucleic acid molecules, which encode any one of the antibodies or its antigen-binding portion thereof that can be used for methods disclosed herein. Nucleic acid can exist in intact cells, cell lysates or partially purified or substantially pure form. When purified from other cellular components or other contaminants (for example, other cellular nucleic acids (for example, other chromosomal DNA, for example, the chromosomal DNA connected with the isolated DNA in nature) or protein by standard techniques (including alkaline / SDS treatment, CsCl banding, column chromatography, restriction enzymes, agarose gel electrophoresis and other technologies well known in the art), nucleic acid is "separated" or "becomes substantially pure". See, F.Ausubel et al., editors (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York. Nucleic acid as described herein can be, for example, DNA or RNA, and can contain or not contain intron sequences. In certain embodiments, nucleic acid is a cDNA molecule.

[0300] Nucleic acids as described herein can be obtained using standard molecular biology techniques. For antibodies expressed by hybridomas (e.g., hybridomas prepared by transgenic mice carrying human immunoglobulin genes, as further described below), cDNAs encoding the light and heavy chains of antibodies prepared by hybridomas can be obtained by standard PCR amplification or cDNA cloning techniques. For antibodies obtained from immunoglobulin gene libraries (e.g., using phage display technology), nucleic acids encoding antibodies can be recovered from the libraries.

[0301] Certain nucleic acid molecules described herein are those encoding the VH and VL sequences of anti-FAM19A5 antibodies of the present disclosure. Exemplary DNA sequences encoding the VH sequences of such antibodies are shown in SEQ ID NOs: 43-46, 177, 205, 235-243, 255, 258, 273, and 276 (see Table 6). Exemplary DNA sequences encoding the VL sequences of such antibodies are shown in SEQ ID NOs: 47-50, 178, 206, 244-252, 256, 259, 274, and 277 (see Table 7).

[0302] Table 6: Variable heavy chain polynucleotide sequences

[0303]

[0304]

[0305]

[0306]

[0307]

[0308] Table 7: Variable light chain polynucleotide sequences

[0309]

[0310]

[0311]

[0312]

[0313] Methods for preparing anti-FAM19A5 antibodies or antigen-binding fragments thereof as disclosed herein can include expressing the relevant heavy and light chains of the antibodies in a cell line comprising nucleotide sequences encoding the heavy and light chains with signal peptides, e.g., SEQ ID NOs: 43 and 47, SEQ ID NOs: 44 and 48, SEQ ID NOs: 45 and 49, SEQ ID NOs: 46 and 50, SEQ ID NOs: 177 and 178, respectively. Host cells comprising these nucleotide sequences are included herein.

[0314] Once the DNA fragments encoding the VH and VL segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example to convert the variable region genes into full-length antibody chain genes, Fab fragment genes or scFv genes. In these manipulations, the DNA fragment encoding VL or VH is operably linked to another DNA fragment encoding another protein, such as an antibody constant region or a flexible linker. As used in this context, the term "operably linked" means that the two DNA fragments are linked together so that the amino acid sequences encoded by the two DNA fragments remain in frame.

[0315] The DNA encoding the separation of VH regions can be converted into full-length heavy chain genes by operably linking the DNA encoding VH to another DNA molecule encoding the heavy chain constant region (hinge, CH1, CH2 and / or CH3). The sequences of human heavy chain constant region genes are known in the art (see, e.g., Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments comprising these regions can be obtained by standard PCR amplification. The heavy chain constant region can be IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, such as IgG2 and / or IgG4 constant region. For Fab fragment heavy chain genes, the DNA encoding VH can be operably linked to another DNA molecule encoding only the heavy chain CH1 constant region.

[0316] By operably connecting the DNA encoding VL to another DNA molecule encoding the constant region of light chain CL, the DNA encoding the separation in the VL region can be converted into full-length light chain genes (and Fab light chain genes). The sequence of human light chain constant region genes is known in the art (see, e.g., Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No.91-3242), and the DNA fragments comprising these regions can be obtained by standard PCR amplification. The constant region of light chain can be a kappa or lambda constant region.

[0317] To generate an scFv gene, the DNA fragments encoding VH and VL are operably linked to another fragment encoding a flexible linker (e.g., encoding the amino acid sequence (Gly4-Ser)3) so that the VH and VL sequences can be expressed as a continuous single-chain protein in which the VL and VH regions are connected by the flexible linker (see, e.g., Bird et al., (1988) Science 242:423-426; Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; McCafferty et al., (1990) Nature 348:552-554).

[0318] In some embodiments, the present disclosure provides a vector comprising an isolated nucleic acid molecule comprising a nucleotide sequence encoding an antibody or an antigen-binding portion thereof. In other embodiments, the vector can be used for gene therapy.

[0319] Suitable vectors for use in the present disclosure include expression vectors, viral vectors, and plasmid vectors. In some embodiments, the vector is a viral vector.

[0320] As used herein, an expression vector refers to any nucleic acid construct that contains the necessary elements for transcription and translation of an inserted coding sequence, or in the case of an RNA viral vector, contains the necessary elements for replication and translation when introduced into a suitable host cell. Expression vectors can include plasmids, phagemids, viruses, and derivatives thereof.

[0321] VI. Antibody Production

[0322] The anti-FAM19A5 antibodies or antigen-binding fragments thereof disclosed herein can be produced by any method known in the art for synthesizing antibodies, such as by chemical synthesis or by recombinant expression techniques. Unless otherwise indicated, the methods described herein employ conventional techniques in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and related fields of the art. These techniques are described, for example, in the references cited herein and are fully explained in the literature. See, for example, Maniatis T et al., (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Sambrook J et al., (1989), Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press; Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel FM et al., Current Protocols in Molecular Biology, John Wiley & Sons (1987 and annual updates); Current Protocols in Immunology, John Wiley & Sons (1987 and annual updates) Gait (ed.) (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press; Eckstein (ed.) (1991) Oligonucleotides and Analogues: A Practical Approach, IRL Press; Birren B et al., (eds.) (1999) Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press.

[0323] In some embodiments, antibodies useful in the methods disclosed herein are antibodies (e.g., recombinant antibodies) prepared, expressed, produced, or isolated by any means involving, for example, synthesis, genetic engineering of a DNA sequence. In certain embodiments, such antibodies comprise sequences (e.g., DNA sequences or amino acid sequences) that are not naturally present in the antibody germline repertoire of an animal or mammal (e.g., a human).

[0324] VII. Pharmaceutical Compositions

[0325] Provided herein are compositions comprising antibodies or antigen-binding portions thereof useful in the methods described herein, the compositions having the desired degree of purity in a physiologically acceptable carrier, excipient, or stabilizer (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl alcohol, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol, and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., zinc protein complexes); and / or nonionic surfactants such as TWEEN®. TM 、PLURONICS TM or polyethylene glycol (PEG).

[0326] In a specific embodiment, the pharmaceutical composition comprises an antibody or antigen-binding portion thereof, bispecific molecule, or immunoconjugate described herein, and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable carrier. In a specific embodiment, the pharmaceutical composition comprises an effective amount of an antibody or antigen-binding portion thereof, and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable carrier. In some embodiments, the antibody is the only active ingredient contained in the pharmaceutical composition. The pharmaceutical compositions described herein can be used to enhance, induce, or activate FAM19A5 activity and treat conditions such as central nervous system damage, degenerative brain disorders, or neuropathic pain.

[0327] The pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, non-aqueous vehicles, antimicrobials, isotonic agents, buffers, antioxidants, local anesthetics, suspending agents and dispersants, emulsifiers, sequestrants or chelating agents and other pharmaceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose and lactated Ringer's injection. Non-aqueous parenteral vehicles include fixed oils of plant origin, cottonseed oil, corn oil, sesame oil and peanut oil. Antimicrobials at bacteriostatic or antifungal concentrations can be added to parenteral preparations packaged in multidose containers, which include phenol or cresol, mercury, benzyl alcohol, chlorobutanol, methylparaben and propylparaben, thimerosal, benzalkonium chloride and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinyl pyrrolidone. Emulsifiers include polysorbate 80 ( 80). Metal ion sequestrants or chelators include EDTA. Pharmaceutical carriers also include ethanol, polyethylene glycol, and propylene glycol for water-miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0328] Pharmaceutical compositions can be formulated for any route of administration to a subject. Specific examples of route of administration include intranasal, oral, parenteral, intrathecal, intracerebroventricular, pulmonary, subcutaneous, intraperitoneal, intravitreal, or intraventricular. Parenteral administration characterized by subcutaneous, intramuscular, or intravenous injection is also contemplated herein. Injectables can be in conventional form (as liquid solutions or suspensions), in solid forms suitable for dissolving or suspending in liquids before injection, or prepared as emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the pharmaceutical composition to be administered can also contain a small amount of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers, stabilizers, solubility enhancers, and other such agents, such as, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins.

[0329] Preparations for parenteral administration of antibodies include sterile solutions ready for injection, sterile dry soluble products (such as lyophilized powders) ready for mixing with solvents before use, including subcutaneous tablets, sterile suspensions ready for injection, sterile dry insoluble products ready for mixing with vehicles before use, and sterile emulsions. The solution can be an aqueous solution or a non-aqueous solution.

[0330] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol and polypropylene glycol, and mixtures thereof.

[0331] Topical mixtures containing the antibody are prepared as described for topical and systemic administration. The resulting mixture can be a solution, suspension, emulsion, or the like, and can be formulated into a cream, gel, ointment, emulsion, solution, elixir, lotion, suspension, tincture, paste, foam, aerosol, rinse, spray, suppository, bandage, skin patch, or any other formulation suitable for topical administration.

[0332] The antibodies described herein, or antigen-binding portions thereof, can be formulated as aerosols for topical administration, such as by inhalation (see, e.g., U.S. Patents Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivering steroids that can be used to treat inflammatory diseases, particularly asthma). These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a fine powder for insufflation, alone or in combination with an inert carrier such as lactose. In such cases, in some embodiments, the particles of the formulation will have a diameter of less than 50 microns, and in some embodiments, less than 10 microns.

[0333] The antibodies described herein, or antigen-binding portions thereof, can be formulated for local or topical administration in the form of gels, creams, and lotions, such as for topical application to the skin and mucous membranes (such as in the eye), as well as for application to the eye or for intracisternal or intraspinal administration. Topical administration is contemplated for transdermal delivery, and also for application to the eye or mucous membranes, or for inhalation therapy. Nasal solutions of the antibodies can also be administered alone or in combination with other pharmaceutically acceptable excipients.

[0334] Transdermal patches (including iontophoresis and electrophoresis devices) are well known to those skilled in the art and can be used to administer antibodies. For example, such patches are disclosed in U.S. Patents 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010715, 5,985,317, 5,983,134, 5,948,433 and 5,860,957.

[0335] In certain embodiments, pharmaceutical compositions comprising an antibody or antigen-binding portion thereof described herein are lyophilized powders that can be reconstituted for administration in the form of solutions, emulsions, and other mixtures. They can also be reconstituted and formulated as a solid or gel. The lyophilized powder is prepared by dissolving an antibody or antigen-binding portion thereof described herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. In some embodiments, the lyophilized powder is sterile. The solvent may contain excipients that enhance the stability of the powder or a reconstituted solution prepared from the powder, or other pharmacological components. Excipients that may be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerol, glucose, sucrose, or other suitable agents. In some embodiments, the solvent may also contain a buffer such as citrate, sodium phosphate, or potassium phosphate, or other such buffers known to those skilled in the art, at a neutral pH. The solution is then sterile filtered and then lyophilized under standard conditions known to those skilled in the art to provide the desired formulation. In some embodiments, the resulting solution is dispensed into vials for lyophilization. Each vial will contain a single or multiple doses of the compound. The lyophilized powder can be stored under appropriate conditions, such as at approximately 4°C to room temperature.

[0336] Reconstitution of this lyophilized powder with water for injection provides a formulation for parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or another suitable carrier. The exact amount depends on the compound selected. Such an amount can be determined empirically.

[0337] Antibodies as herein described or their antigen-binding portion thereof, bispecific molecules or immunoconjugates and other compositions provided herein can also be formulated into specific tissues, receptors or other regions of the body of a target subject to be treated. Many such targeting methods are well known to those skilled in the art. All such targeting methods are contemplated for use in compositions of the present invention herein. For non-limiting examples of targeted approaches, see, e.g., U.S. Patent Nos. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874. In a specific embodiment, the antibodies described herein, or antigen-binding portions thereof, are targeted to treat central nervous system damage, degenerative brain disorders, or neuropathic pain.

[0338] Compositions to be used for in vivo administration may be sterile. This is readily accomplished by, for example, filtration through sterile filtration membranes.

[0339] VIII. Kit

[0340] Provided herein are kits comprising one or more antibodies or antigen-binding portions thereof, bispecific molecules or immunoconjugates thereof that can be used for the methods described herein. In a specific embodiment, provided herein is a pharmaceutical pack or kit comprising one or more containers filled with one or more ingredients of the pharmaceutical compositions described herein (such as one or more antibodies or antigen-binding portions thereof provided herein, and optional instructions for use). In some embodiments, the kit contains the pharmaceutical compositions described herein and any prophylactic or therapeutic agent, such as those described herein.

[0341] Examples

[0342] The following examples are offered by way of illustration and not limitation.

[0343] Example 1 Expression and purification of human FAM19A5 protein

[0344] Recombinant human FAM19A5 protein was produced and purified as described below, and the purified protein was used for antibody screening assays based on binding affinity analysis. First, the LPS-hT plasmid expressing the FAM19A5 gene was transformed into bacteria, and overexpression of the protein was induced. Once produced, the FAM19A5 protein was purified using Ni-NTA affinity chromatography (Qiagen, Valencia, California, USA). Using gradually increasing concentrations of imidazole, we removed the His-tagged FAM19A5 protein from the Ni-column. Protein expression in the solution was measured using Coomassie Brilliant Blue R-250 dye. Only solutions containing FAM19A5 imidazole were used, and the FAM19A5 protein was concentrated using PBS. When concentration was complete, the purity and concentration of the FAM19A5 protein were measured using Western blotting. The concentrated protein was subsequently used to screen for FAM19A5-specific antibodies.

[0345] Example 2 Generation of Antibody Library FAM19A5

[0346] 1. Immunity

[0347] Synthetic FAM19A5 peptide, conjugated to KLH (Anygen) at the C-terminus, and used as the antigen for the immunity of chicken.50 μ L of synthetic peptide (VTLDRDSSQPRRTIARQT) KLH conjugate (SEQ ID NO:262) was mixed in 750 μ L of phosphate buffered saline (PBS) and incubated at 37 ° C for 30 minutes. Then, in the 2% squalene endotoxin MPL (monophosphate lipid A species) and mycobacteria (mycobacteria) of the cell wall component of TDW and CWS containing emulsified oil-in-water emulsion adjuvant (RIBI+MPL+TDM+CWS adjuvant, Sigma, St.Louis, Mo, USA), toxin was removed and then subcutaneously injected into three chickens. These chickens were immunized three times in total, with an interval of about 2-3 weeks between immunizations. The titer of the antibody obtained from the immunized chicken was measured by immunoblotting using the lysate of HEK293T cells overexpressing FAM19A5 protein. Serum from chickens that received three immunizations was used as the primary antibody. The secondary antibody used was an anti-chicken IgG(γ) polyclonal antibody conjugated to HRP (horseradish peroxidase) (rabbit anti-chicken IgG(γ)-HRP, Millipore Corporation, Billerica, MA, USA).

[0348] 2. Preparation of single-chain variable fragment (scFv) libraries from immunized chickens

[0349] We extracted RNA from the spleen, bone marrow, and synovial bursa of the immunized chickens described above using TRI reagent (Invitrogen, Carlsbad, CA, USA). TM The first-chain cDNA is synthesized by using the PCR amplification system (Invitrogen) PCR. For the cDNA obtained from the immune system of chicken, the single-chain variable region library is generated using the Expand High Fidelity PCR System (Roche Molecular Systems, Indiana, USA). In each reaction, 1 μ L of cDNA, 60 pmol of every kind of primer, 10 μ L of 10 × reaction buffer, 8 μ L of 2.5 mM dNTP (Promega, Madison, Wisconsin, USA) and 0.5 μ L of Taq DNA polymerase are mixed with water. The final volume is 100 μ L, and the PCR reaction is carried out under the following conditions: (i) 15 seconds at 94 ° C (ii) 30 seconds at 56 ° C and (iii) 30 cycles of 90 seconds at 72 ° C, followed by final extension for 10 minutes at 72 ° C. The PCR product containing a fragment of approximately 350 bp in length was loaded onto a 1.5% agarose gel and, after electrophoresis, the nucleotide fragments were purified using a QIAGEN Gel II extraction kit (QIAGEN, Valencia, CA, USA). The purified PCR product was quantified by reading at OD 260 nm. (1 unit OD = 50 μL / mL).

[0350] The two VH and VL first products from the second PCR are randomly connected by overlap extension PCR (Overlap extension PCR). Each PCR reaction is mixed with 100ng of the purified VL and VH products, 60pmol of each primer, 10 μL of 10 × reaction buffer, 8 μL of 2.5mM dNTP, 0.5 μL of Taq DNA polymerase and water, and the final volume is 100. PCR is carried out under the following conditions: (i) at 94°C for 15 seconds, (ii) at 56°C for 30 seconds and (iii) at 72°C for 2 minutes for 25 cycles, followed by final extension for 10 minutes at 72°C. The PCR product comprising the single-chain variable region fragment of about 700bp in length is loaded onto a 1.5% agarose gel, and after electrophoresis, the nucleotide fragment is purified using QIAGEN II gel extraction kit (Qiagen). The PCR product of quantitative purification is read at OD 260nm. (1 unit OD=50 / ml).

[0351] 3.Library, Ligation, and Transformation

[0352] The scFv fragment of the PCR product and the vector pComb3X-SS (The Scripps Research Institute, CA, USA) were digested with Sfi I restriction enzyme. 10 μg of the purified overlapping PCT product was mixed with 360 units of Sif I (per 16 units of μg DNA, Roche Molecular Systems, Pleasanton, CA, USA), 20 μL of 10× reaction buffer, and water to a final volume of 200 μL. 20 μg of the pComb3X-SS vector was mixed with 120 units of Sfi I (per 6 units of μg DNA), 20 μL of 10× reaction buffer, and water to a final volume of 200 μL. The mixture was digested at 50°C for 8 hours. The digestion product containing the scFv fragment (approximately 700 bp) and the vector (approximately 3400 bp) was then loaded onto a 1% agarose gel and purified using a Gel Extraction Kit II QIAGEN (QIAGEN, Valencia, CA, USA). 1400 ng of SfiI-restricted pComb3X vector and 700 ng of the digested scFv fragment were mixed with 5× ligase buffer, 10 μL of T4 DNA ligase (Invitrogen, Carlsbad, CA, USA) and water to a final volume of 200 μL. The mixture was incubated at 16°C for 16 hours for ligation.

[0353] After precipitating with ethanol, the DNA precipitate was dissolved in 15 μL of water. In order to produce the library, the connection sample was transformed into Escherichia coli strain ER2738 (New England Biolabs Inc, Hitchin SG4 OTY, Hertfordshire, England, UK) by electroporation using a vibrator gene (Gene Pulser: Bio-Rad Laboratories, Hercules, California, USA). The cells were mixed in 5ml of super broth (SB) culture medium and incubated for one hour at 37°C with 250rpm stirring. Then, 3 μL of 100mg / mL of kanamycin was added to 10mL of SB culture medium. In order to determine the library size, 0.1 μL, 1 μL and 10 μL of culture samples were applied to Luria broth (LB) agar plates containing 50 μg / ml of kanamycin. After stirring for 1 hour, 4.5 μL of 100mg / mL of kanamycin were added to the LB culture, and further stirred for another 1 hour. Then, 2 ml of VCM13 helper phage (>10 11cfu / ml) together with the preheated LB (183mL) of the 100mg / mL kanamycin containing 92.5 μ L are added in LB substratum.This mixture was stirred other 2 hours at 37 ℃ with 250rpm.Then, the kanamycin of 280 μ L (50mg / mL) is added in the culture, and stirred overnight at 37 ℃.The next day, high-speed centrifuge (Beckman, JA-10 rotor) is used to centrifuge bacteria precipitation at 4 ℃ with 3,000g.Afterwards, the bacteria precipitation is used to extract phagemid DNA, and the supernatant is transferred to a sterile centrifuge bottle simultaneously.Next, the polyethylene glycol-8000 (PEG-8000, Sigma (Sigma)) of 8g and the sodium chloride (NaCl, Merck (Merck)) of 6g are added in the supernatant, then kept 30 minutes in ice.Then, the supernatant is centrifuged 15 minutes at 4 ℃ with 15,000g. The supernatant was then discarded, and the pelleted phages were suspended in Tris-buffered saline (TBS) containing 1% BSA.

[0354] Example 3: Library panning on immobilized antigen (biopanning)

[0355] Biopanning was performed using magnetic beads (Dynabeads M-270 Epoxy, Invitrogen). The beads and protein were stirred together at room temperature for 20 hours while rotating. 7 Each bead was coated with 5 μg of recombinant FAM19A5 protein. Once the coating was completed, the beads were washed 4 times with phosphate buffered saline (PBS) and blocked for one hour at room temperature in PBS containing 3% BSA. Then, the coated beads were cultured at room temperature for two hours with the scFv displayed by the phage. In order to remove any phage that was not bound to the antigen-coated beads, the beads were washed with 0.05% Tween 20 / PBS. The bound phage was then eluted with 50 μL of 0.1M glycine / hydrogen chloride (0.1M glycine-HCl, pH 2.2) and neutralized with 3 μL of 2M Tris (tris-HCl, pH 9.1) containing hydrogen chloride. This phage-containing supernatant was used to infect Escherichia coli ER2738 cells and amplified and rescued overnight using VCSM13 helper phage. In addition, by the culture of phage infection on the LB agar plate containing the kanamycin of 50 μ g / mL, trace is determined the input (input) and the output (output) of the phage titer from the culture of phage infection.On the second day, PEG-8000 and NaCl are used to precipitate phage, subsequently it is used for biological panning.By repeating said process, carried out five biological pannings altogether.Under the situation of each amplification, screen phage and select the high-affinity to FAM19A5 protein.

[0356] Example 4 Clone selection by phage ELISA

[0357] To analyze clones selected from biopanning, we randomly selected individual clones from the phage-displayed scFvs and confirmed their binding to FAM19A5 recombinant protein using ELISA. FAM19A5 recombinant protein was diluted in 0.1 M NaHCO3 buffer, and 100 ng / well of protein was used to coat 96-well microtiter plates at 4°C for 16 hours. The next day, the plates were blocked with 3% BSA / PBS at 37°C for 1 hour. Phage supernatants were then mixed with 6% BSA / PBS and incubated at 37°C for 2 hours. The plates containing the supernatants were then washed with 0.05% Tween 20 / PBS. The HRP-conjugated M13 antibody (a-M13-HRP, Pierce Chemical Company, Rockford, IL, USA) was diluted to 1 / 5000. 50 μl of the diluted antibody was added to the plate and incubated at 37°C for 1 hour. After incubation and washing, 0.05 M citrate buffer, 1 μg / mL 2,2'-azido-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS, Amresco, Solon, OH, USA) and 0.1% H2O2 were added to the plate for color development. The absorbance of each well was measured at 405 nm. Figure 1A 、 1B A and 1C show the analysis of 96 clones from level 3, 4, or 5 biopanning of the first, second, and third chickens.

[0358] As in Figures 1A to 1C As shown in , we analyzed 24 clones that bound to FAM19A5 recombinant protein and showed high absorbance, and from these 24 clones, we obtained 13 scFv clones with unique sequences. After further selection of clones, we obtained clone 1-65 with the highest affinity.

[0359] Example 5 Generation of anti-FAM19A5-IGG2 / 4 antibodies

[0360] 1. Subcloning of anti-FAM19A5 scFv into mammalian expression vector

[0361] In the FAM19A5 scFv gene sequence, the human Cκ gene is connected to the light chain variable domain, and the human immunoglobulin isotype IgG2 / 4 of the CH1, CH2 and CH3 genes is connected to the heavy chain variable region. Antibodies with each light chain and each heavy chain are synthesized by adding restriction sites (Genscript, USA). The synthesized genes are inserted into a mammalian cell expression vector with modified restriction sites to facilitate cloning. First, the light chain genes are inserted into the vector using Hind III and Xba I (New England Biolabs, UK) restriction enzymes, and then the heavy chain genes are added to the vector using NheI and BamHI (New England Biolabs, UK) restriction enzymes. Figure 2 ).

[0362] 2. Purification of Anti-FAM19A5 Antibodies

[0363] To express and purify anti-FAM19A5-IgG2 / 4 antibodies, we used a mammalian cell transfection and overexpression injection system. We mixed 2 μg / mL of mammalian expression vector with 4 μg of polyethyleneimine (PEI, Polysciences, Warrington, PA, USA) in 150 mM sodium chloride (NaCl, Merck), equivalent to 1 / 10 of the cell culture volume. The mixture was allowed to stand at room temperature for 15 minutes. The mixture was added to HEK293F cells (2 × 10 6 cells / ml, Invitrogen) and then in FREESTYLE containing 100 U / ml of penicillin and streptomycin (Invitrogen). TM The cells were incubated in 293 expression medium at 7% CO2 and 37°C with agitation at 135 rpm for six days. To purify the expressed anti-FAM19A5 IgG2 / 4 antibody from the cell culture supernatant, we used affinity gel chromatography with protein A beads (RepliGen, Waltham, MA, USA). The protein A chromatography-purified antibody was run on a 4-12% Bis-Tris gradient gel electrophoresis. The size and yield of the protein were confirmed by Coomassie brilliant blue staining ( Figure 3 ).

[0364] Example 6 Evaluation of Mechanical Hyperalgesia Following In Vivo Administration of Anti-FAM19A5 Antibodies in a Rat Model of Chronic Constriction Injury

[0365] To investigate whether neutralization of FAM19A5 activity in vivo can alleviate neuropathic pain, a rat model of chronic constriction injury (CCI) was used, as previously described by Bennett and Xie, Pain 33(1):87-107(1988), Austin et al., J Vis Exp 61:3393(2012). Experimental CCI of the sciatic nerve is one of the most widely used models for studying neuropathic pain and is reported to induce an inflammatory response in the ipsilateral hind paw. Therefore, the hind paw withdrawal threshold, as measured, for example, by the von Frey test, is used as a good indicator of neuropathic pain.

[0366] 1. Inducing neuropathic pain through chronic constriction injury

[0367] Briefly, Zoletil 50 (Vick, France) and xylazine ( , Bayer AG, Germany) IP (intraperitoneal injection) is used to deeply anesthetize 6-week-old male Sprague-Dawley rats. Then, the hair of the lower back and thigh of the rat is shaved, and the skin is disinfected with povidone iodine. Next, the skin of the lateral surface of the thigh is incised, and the sciatic nerve is exposed at the middle thigh level by blunt incision via the biceps femoris. At the proximal end of the trifurcation of the sciatic nerve, about 7mm of the nerve has no adherent tissue, and 4 ligatures (4.0 black silk) are loosely tied around it at intervals of about 1mm. The length of the nerve thus affected is 4-5mm long. After performing nerve ligation, the muscle layer and skin layer are sutured in layers with a thread, and local antibiotics are used.

[0368] 2. Administration of Anti-FAM19A5 Antibodies

[0369] Zoletil 50 (Vick, France) and xylazine ( Male Sprague-Dawley rats were anesthetized with human IgG from a human IgG4-positive rat model (Hyperbactam, Bayer, Germany) and divided into four groups as shown in Table 8. One group of rats with chronic constriction injury ("CCI-induced rats" (G3, n = 10) received anti-FAM19A5 antibody (10 μg / rat in a volume of 0.1 mL) by intrathecal injection. The antibody was administered once a week for 2 weeks (i.e., on days 7 and 14 after CCI induction). Another group of CCI-induced rats (G2, n = 10) served as a "negative control" and received only human IgG. Another group of CCI-induced rats (G4, n = 10) were treated with pregabalin, a drug used to treat neuropathic pain. The remaining group of rats (G1, n = 10) served as a "sham control" (i.e., no CCI induction and no drug administration).

[0370] Table 8

[0371]

[0372] 3. Von Frey Test

[0373] On days 6 and 21 after chronic constriction of the sciatic nerve, the paw withdrawal threshold was assessed using the von Frey test. Rats were placed in an apparatus with a metal mesh floor and allowed to stabilize in the environment for approximately 20 minutes. The paw withdrawal threshold was then measured by applying a von Frey filament (0.5 mm diameter) to the plantar surface of the hind paw three times at 10-second intervals through the mesh floor.

[0374] result

[0375] like Figure 5A and 5B As shown, in vivo administration of anti-FAM19A5 antibodies after CCI induction resulted in a significant increase in paw withdrawal thresholds. Early after administration (e.g., day 6), there were no significant differences between the groups. However, around day 21 after chronic constriction of the sciatic nerve, significant differences between the treated groups (i.e., anti-FAM19A5 antibodies (G3) or pregabalin (G4)) and the negative control group (G2) were significant. For example, on day 6, the mean paw withdrawal thresholds for the negative control (G2) and anti-FAM19A5 antibody-treated rats (G3) were 5.1 ± 1.0 and 6.6 ± 2.0, respectively. Figure 5A However, on day 21 after chronic constriction of the sciatic nerve, the mean threshold of the negative control (G2) was 5.5±1.3, while for the anti-FAM19A5 antibody-treated group (G3), the mean threshold was 8.5±1.7 (p<0.01), which was comparable to the mean threshold of pregabalin (G4: 8.5±1.7), a drug used to treat neuropathic pain. Figure 5B These results suggest that neutralizing FAM19A5 activity by administering anti-FAM19A5 antibodies in vivo can ameliorate neuropathic pain.

[0376] Example 7 Evaluation of motor function after in vivo administration of anti-FAM19A5 antibodies

[0377] In order to study whether anti-FAM19A5 antibody treatment can change motor function, the rotarod test was used to assess the motor activity of rats induced by CCI. This test is used as a good indicator of any pain or muscle weakness in the lower limbs caused by CCI induction (Chen L et al., 2014, Vadakkan KI et al., 2005). CCI induction and anti-FAM19A5 antibody administration were performed as described in Example 1.

[0378] 1. Rotarod test

[0379] Each Sprague-Dawley rat (sham control and CCI-induced) was carefully placed on a rotarod (Biological Research Apparatus 7750, UGO BASILE, Italy), and the rotation speed of the rotarod was increased from 4 rpm to 20 rpm at regular intervals. The latency to fall was recorded on day 6 (baseline) and day 21 after administration of anti-FAM19A5 antibodies. Motor performance was considered to be the latency to fall from the rotarod apparatus, which was determined based on the average time of three trials per rat.

[0380] 2. Results

[0381] like Figure 6A and 6B As shown, in vivo administration of anti-FAM19A5 antibodies after CCI induction resulted in improved latency times (i.e., rats stayed on the rotarod apparatus longer). Although there were no significant differences between the groups on day 6 after chronic constriction of the sciatic nerve ( Figure 6A ), but by day 21, rats treated with anti-FAM19A5 antibody showed a much longer latency compared with the negative control (G2: 64.3 ± 14.1 vs. G3: 92.8 ± 11.5), which was comparable to the latency of pregabalin (G4: 91.1 ± 17.8) ( Figure 6B These results suggest that in vivo administration of anti-FAM19A5 antibodies after CCI induction can not only alleviate neuropathic pain but also improve motor function.

[0382] Example 8: Evaluation of mechanical hyperalgesia following in vivo administration of anti-FAM19A5 antibodies in a rat model of diabetic peripheral neuropathy

[0383] In order to further evaluate the benefits of neutralizing FAM19A5 activity to neuropathic pain, a rat model of diabetic peripheral neuropathy was used. By administering streptozotocin (STZ) (50 mg / kg, intraperitoneally) to animals, diabetic peripheral neuropathy (DPN) was induced in Sprague-Dawley rats. Blood glucose levels were measured approximately one week after STZ administration, and only animals with fasting blood glucose levels of 300 mg / dL or higher were selected for antibody administration. Approximately 3 weeks after STZ administration, selected DPN rats received control antibodies (normal human IgG, "NHI") or anti-FAM19A5 antibodies (3-2) administered weekly (a total of 8 times, i.e., from the 4th week to the 11th week after STZ administration). Antibodies were administered intrathecally to animals at a dosage of 25 g / rats. Initial (healthy) animals were used as "sham controls" (i.e., no DPN induction and no antibody administration). Paw withdrawal thresholds were measured in animals using the von Frey test as described in Example 6 at 7 and 11 weeks after STZ administration (ie, 4 and 8 weeks after the start of antibody administration, respectively).

[0384] like Figure 10 As shown, DPN animals that received anti-FAM19A5 antibodies had significantly higher paw withdrawal thresholds than those of DPN animals that received control antibodies. This increase in threshold was observed at both the 7th and 11th weeks after STZ administration (i.e., the 4th and 8th weeks after antibody administration, respectively).

[0385] Example 9: Evaluation of thermal hyperalgesia following in vivo administration of anti-FAM19A5 antibodies in a rat model of diabetic peripheral neuropathy

[0386] Next, to evaluate the effect of anti-FAM19A5 administration on neuropathic pain associated with thermal hyperalgesia, diabetic peripheral neuropathy (DPN) was induced in Sprague-Dawley rats, and control (NHI) or anti-FAM19A5 antibody (3-2) was administered to the animals as described above in Example 8. Then, at 4 and 8 weeks after the start of antibody administration (i.e., 7 and 11 weeks after STZ administration), the animals were placed on a hot plate and the temperature was immediately raised to 55° C. The time it took for the animals to respond to the increased temperature (e.g., by jumping or licking their paws) was then measured.

[0387] like Figure 11A and 11BAs shown, DPN animals receiving a control antibody were significantly more sensitive to heat stimulation, with a paw withdrawal latency of approximately 6 seconds. In contrast, DPN rats receiving an anti-FAM19A5 antibody had significantly higher paw withdrawal latencies at both weeks 4 and 8. These data further demonstrate that neutralizing FAM19A5 activity can improve neuropathic pain associated with mechanical and thermal hyperalgesia.

[0388] Example 10 Analysis of sensory nerve conduction velocity after in vivo administration of anti-FAM19A5 antibodies in a rat model of diabetic peripheral neuropathy

[0389] In order to determine the effect of anti-FAM19A5 administration on peripheral sensory nerve damage and / or dysfunction, sensory nerve conduction velocity (SNCV) test was carried out. By measuring the speed at which electrical signals are transmitted from descending sensory nerves to ascending sensory nerves, this test can help identify potential nerve damage and / or dysfunction. As described in Example 8, diabetic peripheral neuropathy was induced in Sprague-Dawley rats using STZ, and controls or anti-FAM19A5 antibodies (3-2) were administered to animals. Then, 8 weeks after starting antibody administration, the nerve conduction velocity in the sural nerve was measured. Viking Quest (Natus Neurology Incorporated, U.S.) equipment was used to induce electrical stimulation at 10 mA and 10 μV.

[0390] like Figure 12 As shown, DPN rats that received anti-FAM19A5 antibodies had improved conduction velocity compared to those DPN animals that received control antibodies. This result suggests that neutralizing FAM19A5 activity can improve and / or reduce peripheral nerve damage / dysfunction, thereby helping to alleviate neuropathic pain, as observed in the previous examples. Overall, the above results suggest that FAM19A5 antagonists (such as the anti-FAM19A5 antibodies disclosed herein) may be a suitable treatment option for neuropathic pain.

[0391] Example 11 Epitope Mapping

[0392] Overlapping peptide fragments of human FAM19A5 protein (F1-F5, see Figure 9) and conjugated to BSA. The binding of monoclonal antibody 1-65 to BSA-conjugated peptide fragments F1-F6 was determined by Western blot analysis. For Western blot analysis, BSA-conjugated FAM19A5 fragments F1-F6 were separated by SDS polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes by standard procedures. The membranes were incubated with anti-FAM19A5 antibody 1-65 (2 μg / ml, 1-65-ScFv-rabbit Fc-SSS), and the antigen-antibody complexes were detected with a suitable secondary antibody conjugated to horseradish peroxidase (anti-rabbit IgG (Fc specific)-HRP, 1:4000 dilution). Figure 9 ).like Figure 9 As shown, anti-FAM19A5 antibody 1-65 strongly binds to fragment F5. In addition to F5, it also binds to fragments F2 and F3.

[0393] It should be understood that the Detailed Description section, and not the Summary and Abstract sections, is intended to be used to interpret the claims. The Summary and Abstract sections may set forth one or more, but not all, exemplary embodiments of the present disclosure as contemplated by the inventors, and thus, are not intended to limit the present disclosure and the appended claims in any way.

[0394] The present disclosure has been described with the aid of functional building blocks that illustrate the implementation of specified functions and their relationships. For ease of description, the boundaries of these functional building blocks have been arbitrarily defined in this article. Alternative boundaries can be defined as long as the specified functions and their relationships are appropriately performed.

[0395] The foregoing description of the specific embodiments will reveal the general nature of the present disclosure so fully that others can easily modify and / or adapt various applications, such as the specific embodiments, by applying knowledge within the art without departing from the general concepts of the present disclosure, without undue experimentation. Therefore, based on the teachings and guidance given herein, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed embodiments. It should be understood that the wording or terminology herein is for descriptive purposes only and not for limiting purposes, so that the terms or wording of this specification will be interpreted by the skilled person in accordance with the teachings and guidance.

[0396] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0397] All publications, patents, patent applications, internet sites, and accession / database sequences (including polynucleotide and polypeptide sequences) cited herein are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, internet site, or accession / database sequence was specifically and individually indicated to be incorporated by reference.

[0398] This PCT application claims the benefit of priority to U.S. Provisional Application No. 62 / 661,923, filed April 24, 2018, which is incorporated herein by reference in its entirety. Sequence Listing <110> NEURACLE SCIENCE CO., LTD. <120> Use of antibodies against sequence similarity family 19 member A5 in the treatment of neuropathic pain <130> 3763.009PC01 / CK / DKC <150> US 62 / 661,923 <151> 2018-04-24 <160> 277 <170> PatentIn Version 3.5 <210> 1 <211> 132 <212> PRT <213> Homo sapiens <400> 1 Met Ala Pro Ser Pro Arg Thr Gly Ser Arg Gln Asp Ala Thr Ala Leu 1 5 10 15 Pro Ser Met Ser Ser Thr Phe Trp Ala Phe Met Ile Leu Ala Ser Leu 20 25 30 Leu Ile Ala Tyr Cys Ser Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 35 40 45 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 50 55 60 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 65 70 75 80 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 85 90 95 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 100 105 110 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 115 120 125 Thr Thr Val Ser 130 <210> 2 <211> 125 <212> PRT <213> Homo sapiens <400> 2 Met Gln Leu Leu Lys Ala Leu Trp Ala Leu Ala Gly Ala Ala Leu Cys 1 5 10 15 Cys Phe Leu Val Leu Val Ile His Ala Gln Phe Leu Lys Glu Gly Gln 20 25 30 Leu Ala Ala Gly Thr Cys Glu Ile Val Thr Leu Asp Arg Asp Ser Ser 35 40 45 Gln Pro Arg Arg Thr Ile Ala Arg Gln Thr Ala Arg Cys Ala Cys Arg 50 55 60 Lys Gly Gln Ile Ala Gly Thr Thr Arg Ala Arg Pro Ala Cys Val Asp 65 70 75 80 Ala Arg Ile Ile Lys Thr Lys Gln Trp Cys Asp Met Leu Pro Cys Leu 85 90 95 Glu Gly Glu Gly Cys Asp Leu Leu Ile Asn Arg Ser Gly Trp Thr Cys 100 105 110 Thr Gln Pro Gly Gly Arg Ile Lys Thr Thr Thr Val Ser 115 120 125 <210> 3 <211> 53 <212> PRT <213> Homo sapiens <400> 3 Met Tyr His His Arg Glu Trp Pro Ala Arg Ile Ile Lys Thr Lys Gln 1 5 10 15 Trp Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu 20 25 30 [[ID=3三十三]]Ile Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys 35 40 45 Thr Thr Thr Val Ser 50 <210> 4 <211> 1501 <212> DNA <213> Homo sapiens <400> 4 ggcggcggag gatggcgcgc gcggggcccg cacgtggagg ccggcgcggg ggcgcgggca 60 It should be noted that in the translation of line 33, there is an extra "三十三" which seems to be an error in the original text. It is translated as "Ile Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys" as it is likely a misprint in the original and this is the most reasonable way to handle it for translation. If this is not an error, please provide more context or clarify the specific requirements for that part.gggccggctg ctgagacgcg ctgctgcccc ccgcgcgggc gccgcggctt caatggcgcc 120 atcgcccagg accggcagcc ggcaagatgc gaccgccctg cccagcatgt cctcaacttt 180 ctgggcgttc atgatcctgg ccagcctgct catcgcctac tgcagtcagc tggccgccgg 240 cacctgtgag attgtgacct tggaccggga cagcagccag cctcggagga cgatcgccccg 300 gcagaccgcc cgctgtgcgt gtagaaaggg gcagatcgcc ggcaccacga gagcccggcc 360 cgcctgtgtg gacgcaagaa tcatcaagac caagcagtgg tgtgacatgc ttccgtgtct 420 ggaggggaa ggctgcgact tgttaatcaa ccggtcaggc tggacgtgca cgcagcccgg 480 cgggaggata aagaccacca cggtctcctg acaaacacag cccctgaggg ggccccggga 540 gtggccttgg ctccctggag agcccacgtc tcagccacag ttctccactc gcctcggact 600 tcaccccgttc tctgccgccc gcccatccg tttccctgtg gtccgtgaag gacggcctca 660 ggccttggca tcctgagctt cggtctgtcc agccgacccg aggaggccgg actcagacac 720 ataggcgggg ggcggcacct ggcatcagca atacgcagtc tgtgggagcc cggccgcgcc 780 cagcccccgc cgaccgtggc gttggccctg ctgtcctcag aggagga ggaggaggca 840 gctccggcag ccacagaagg ctgcagccca gcccgcctga ccacgacgc ctgccccagg 900 ggactgtcag gcacagaagc ggcctcctcc cgtgccccag actgtccgaa ttgcttttat 960 tttcttatac tttcagtata ctccatagac caagagcaa atctatctg aacctggacg 1020 caccctcact gtcagggtcc ctggggtcgc tgtcgggc gggagggca tggtggcaga 1080 gatagctgg tggccccggc ggagcggaga gggcggccgt ggtggaggcc tccacccag 1140 gagcaccccg cacaccctcg gaggacgggc tcggctgcg cggaggccgt ggcacctg 1200 cgggaggcag cgacggcccc cacgcagacg ccgggaacgc agggccgcttt attccctgt 1260 acttagatca acttgaccgt actaaaatcc ctttctgttt taaccagtta aacatgccctc 1320 ttctacagct ccatttttga tagttggata atccagtac tgccaagagc atgttgggtc 1380 tcccgtgact gctgcctcat cgatacccca tttagctcca gaaagcaag aaaactcgag 1440 taacacttgt tgaagaga tcattaatg tattttgcaa agcccaaaa aaaaaaaaaaa 1500 and 1501 <210> 5 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Epitope F1 <400> 5 Gln Leu Ala Ala Gly Thr Cys Glu Ile Val Thr Leu Asp Arg 1 5 10 <210> 6 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Epitope F2 <400> 6 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 7 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Epitope F3 <400> 7 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 1 5 10 15 Ala Arg Pro Ala 20 <210> 8 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Epitope F4 <400> 8 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 1 5 10 15 Cys Asp Met Leu 20 <210> 9 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Epitope F5 <400> 9 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 1 5 10 15 Asn Arg Ser Gly 20 <210> 10 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Epitope F6 <400> 10 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 1 5 10 15 Thr Thr Val Ser 20 <210> 11 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 2-13 Antibody VH-CDR1 <400> 11 Ser His Gly Met Phe 1 5 <210> 12 <211> 17 <212> PRT <213> Artificial sequence <220> <223> 2-13 Antibody VH-CDR2 <400> 12 Glu Ile Thr Asn Asp Gly Ser Gly Thr Asn Tyr Gly Ser Ala Val Lys 1 5 10 15 Gly <210> 13 <211> 20 <212> PRT <213> Artificial sequence <220> <223> 2-13 Antibody VH-CDR3 <400> 13 Ser Thr Tyr Glu Cys Pro Gly Gly Phe Ser Cys Trp Gly Asp Thr Gly 1 5 10 15 Gln Ile Asp Ala 20 <210> 14 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 3-2 Antibody VH-CDR1 <400> 14 Ser Phe Asn Met Phe 1 5 <210> 15 <211> 17 <212> PRT <213> Artificial sequence <220> <223> 3-2 Antibody VH-CDR2 <400> 15 Gln Ile Ser Ser Ser Gly Ser Ser Thr Asn Tyr Ala Pro Ala Val Arg 1 5 10 15 Gly <210> 16 <211> twenty two <212> PRT <213> Artificial sequence <220> <223> 3-2 Antibody VH-CDR3 <400> 16 Ser Ser Tyr Asp Cys Pro Tyr Gly His Cys Ser Ser Gly Val Asp Ser 1 5 10 15 Ala Gly Glu Ile Asp Ala 20 <210> 17 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VH-CDR1 <400> 17 Ser Tyr Gln Met Gly 1 5 <210> 18 <211> 10 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VH-CDR2 <400> 18 Val Ile Asn Lys Ser Gly Ser Asp Thr Ser 1 5 10 <210> 19 <211> 13 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VH-CDR3 <400> 19 Gly Ser Ala Ser Tyr Ile Thr Ala Ala Thr Ile Asp Ala 1 5 10 <210> 20 <211> 8 <212> PRT <213> Artificial sequence <220> <223> 1-28 Antibody VH-CDR1 <400> 20 Gly Phe Asp Phe Ser Asp Tyr Gly 1 5 <210> twenty one <211> 8 <212> PRT <213> Artificial sequence <220> <223> 1-28 Antibody VH-CDR2 <400> twenty one Ile Arg Ser Asp Gly Ser Asn Pro 1 5 <210> twenty two <211> 29 <212> PRT <213> Artificial sequence <220> <223> 1-28 Antibody VH-CDR3 <400> twenty two Ala Lys Asp Gly Asn Gly Tyr Cys Ala Leu Asp Ala Tyr Arg Ser Gly 1 5 10 15 Gly Tyr Ser Cys Gly Val Tyr Pro Gly Ser Ile Asp Ala 20 25 <210> twenty three <211> 8 <212> PRT <213> Artificial sequence <220> <223> 2-13 antibody VL-CDR1 <400> twenty three Ser Gly Gly Ser Tyr Ser Tyr Gly 1 5 <210> twenty four <211> 7 <212> PRT <213> Artificial sequence <220> <223> 2-13 antibody VL-CDR2 <400> twenty four Trp Asp Asp Glu Arg Pro Ser 1 5 <210> 25 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 2-13 Antibody VL-CDR3 <400> 25 Gly Thr Glu Asp Ile Ser Gly Thr Ala Gly Val 1 5 10 <210> 26 <211> 13 <212> PRT <213> Artificial sequence <220> <223> 3-2 Antibody VL-CDR1 <400> 26 Ser Gly Gly Gly Ser Tyr Ala Gly Ser Tyr Tyr Tyr Gly 1 5 10 <210> 27 <211> 7 <212> PRT <213> Artificial sequence <220> <223> 3-2 Antibody VL-CDR2 <400> 27 Glu Ser Asn Lys Arg Pro Ser 1 5 <210> 28 <211> 10 <212> PRT <213> Artificial sequence <220> <223> 3-2 Antibody VL-CDR3 <400> 28 Gly Ser Trp Asp Ser Ser Asn Gly Gly Ile 1 5 10 <210> 29 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VL-CDR1 <400> 29 Ser Gly Gly Gly Ser Ser Gly Tyr Gly Tyr Gly 1 5 10 <210> 30 <211> 7 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VL-CDR2 <400> 30 Trp Asn Asp Lys Arg Pro Ser 1 5 <210> 31 <211> 14 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VL-CDR3 <400> 31 Gly Asn Asp Asp Tyr Ser Ser Asp Ser Gly Tyr Val Gly Val 1 5 10 <210> 32 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 1-28 Antibody VL-CDR1 <400> 32 Gly Tyr Gly Tyr Gly 1 5 <210> 33 <211> 3 <212> PRT <213> Artificial sequence <220> <223> 1-28 Antibody VL-CDR2 <400> 33 Gln Asn Asp 1 <210> 34 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 1-28 Antibody VL-CDR3 <400> 34 Gly Ser Glu Asp Ser Ser Thr Leu Ala Gly Ile 1 5 10 <210> 35 <211> 129 <212> PRT <213> Artificial sequence <220> <223> 2-13 Antibody VH <400> 35 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30 Gly Met Phe Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu Tyr Val 35 40 45 Ala Glu Ile Thr Asn Asp Gly Ser Gly Thr Asn Tyr Gly Ser Ala Val 50 55 60 Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Gly Thr Tyr Phe Cys 85 90 95 Ala Arg Ser Thr Tyr Glu Cys Pro Gly Gly Phe Ser Cys Trp Gly Asp 100 105 110 Thr Gly Gln Ile Asp Ala Trp Gly His Gly Thr Glu Val Ile Val Ser 115 120 125 Ser <210> 36 <211> 131 <212> PRT <213> Artificial Sequence <220> <223> 3-2 Antibody VH <400> 36 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30 Asn Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Val 35 40 45 Ala Gln Ile Ser Ser Ser Gly Ser Ser Thr Asn Tyr Ala Pro Ala Val 50 55 60 Arg Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Pro Gly Ala Glu Asp Thr Gly Thr Tyr Tyr Cys 85 90 95 Ala Lys Ser Ser Tyr Asp Cys Pro Tyr Gly His Cys Ser Ser Gly Val 100 105 110 Asp Ser Ala Gly Glu Ile Asp Ala Trp Gly His Gly Thr Glu Val Ile 115 120 125 Val Ser Ser 130 <210> 37 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> VH of Antibody 1-65 [[ID=…]]<400> 37 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gln Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Val Ile Asn Lys Ser Gly Ser Asp Thr Ser Tyr Gly Ser Ala Val 50 55 60 Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Gly Thr Tyr Phe Cys 85 90 95 Ala Lys Gly Ser Ala Ser Tyr Ile Thr Ala Ala Thr Ile Asp Ala Trp 100 105 110 Gly His Gly Thr Glu Val Ile Val Ser Ser 115 120 <210> 38 <211> 136 <212> PRT <213> Artificial Sequence <220> <223> 1-28 antibody VH <400> 38 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Asp Phe Ser Asp Tyr 20 25 30 Gly Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Arg Ser Asp Gly Ser Asn Pro Ser Tyr Gly Ser Ala Val 50 55 60 Lys Gly Arg Ala Thr Ile Ser Lys Asp Asn Gly Arg Ser Thr Val Arg 65 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Lys Asp Gly Asn Gly Tyr Cys Ala Leu Asp Ala Tyr Arg Ser Gly 100 105 110 Gly Tyr Ser Cys Gly Val Tyr Pro Gly Ser Ile Asp Ala Trp Gly His 115 120 125 Gly Thr Glu Val Ile Val Ser Ser 130 135 <210> 39 <211> 104 <212> PRT <213> Artificial Sequence <220> <223> 2 - 13 Antibody VL <400> 39 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu Thr Val 1 5 10 15 Lys Ile Thr Cys Ser Gly Gly Ser Tyr Ser Tyr Gly Trp Phe Gln Gln 20 25 30 Lys Ser Pro Gly Ser Ala Leu Val Thr Val Ile Tyr Trp Asp Asp Glu 35 40 45 Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ala Leu Ser Gly Ser 50 55 60 Thr Asn Thr Leu Thr Ile Thr Gly Val Gln Ala Asp Asp Glu Ala Val 65 70 75 80 Tyr Phe Cys Gly Thr Glu Asp Ile Ser Gly Thr Ala Gly Val Phe Gly 85 90 95 Ala Gly Thr Thr Leu Thr Val Leu 100 <210> 40 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> 3-2 Antibody VL <400> 40 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu Thr Val 1 5 10 15 Lys Ile Thr Cys Ser Gly Gly Gly Ser Tyr Ala Gly Ser Tyr Tyr Tyr 20 25 30 Gly Trp Tyr Gln Gln Lys Ala Pro Gly Ser Ala Pro Val Thr Leu Ile 35 40 45 Tyr Glu Ser Asn Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Thr Ser Gly Ser Thr Ala Thr Leu Thr Ile Thr Gly Val Gln Ala 65 70 ...... Asp Asp Glu Ala Ile Tyr Tyr Cys Gly Ser Trp Asp Ser Ser Asn Gly 85 90 95 Gly Ile Phe Gly Ala Gly Thr Thr Leu Thr Val Leu 100 105 <210> 41 <211> 110 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody VL <400> 41 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu Thr Val 1 5 10 15 Lys Ile Thr Cys Ser Gly Gly Gly Ser Ser Gly Tyr Gly Tyr Gly Trp 20 25 30 Tyr Gln Gln Lys Ser Pro Ser Ser Ala Pro Leu Thr Val Ile Tyr Trp 35 40 45 Asn Asp Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Lys 50 55 60 Ser Gly Ser Thr His Thr Leu Thr Ile Thr Gly Val Gln Ala Glu Asp 65 70 75 80 Glu Ala Val Tyr Phe Cys Gly Asn Asp Asp Tyr Ser Ser Asp Ser Gly 85 90 95 Tyr Val Gly Val Phe Gly Ala Gly Thr Thr Leu Thr Val Leu 100 105 110 <210> 42 <211> 104 <212> PRT <213> Artificial sequence <220> <223> 1-28 antibody VL <400> 42 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Leu Glu Gly Thr Val 1 5 10 15 Glu Ile Thr Cys Ser Gly Ser Gly Tyr Gly Tyr Gly Trp Tyr Gln Gln 20 25 30 Lys Ser Pro Gly Ser Ala Pro Val Thr Val Ile Tyr Gln Asn Asp Lys 35 40 45 Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Lys Ser Gly Ser 50 55 60 Thr Gly Thr Leu Thr Ile Thr Gly Val Gln Val Glu Asp Glu Ala Val 65 70 75 80 Tyr Tyr Cys Gly Ser Glu Asp Ser Ser Thr Leu Ala Gly Ile Phe Gly 85 90 95 Ala Gly Thr Thr Leu Thr Val Leu 100 <210> 43 <211> 388 <212> DNA <213> Artificial sequence <220> <223> 2-13 antibody VH <400> 43 gccgtgacgt tggacgagtc cgggggcggc ctccagacgc ccggaggagc gctcagcctc 60 gtctgcaagg cctccgggtt caccttcagc agccatggca tgttctgggt gcgacagacg 120 cccggcaagg ggttggaata tgtcgctgaa attaccaatg atggtagtgg cacaaactac 180 gggtcggcgg tgaagggccg tgccaccatc tcgagggaca acgggcagag cacagtgagg 240 ctgcagctga acaacctcag ggctgaggac accggcacct acttctgcgc cagatctact 300 tatgaatgtc ctggtggttt tagttgttgg ggtgatactg gtcaaataga cgcatggggc 360 cacgggaccg aagtcatcgt ctcctcca 388 <210> 44 <211> 394 <212> DNA <213> Artificial Sequence <220> <223> 3-2 Antibody VH <400> 44 gccgtgacgt tggacgagtc cgggggcggc ctccagacgc ccggaggagc gctcagcctc 60 gtctgcaagg cctccgggtt caccttcagc agcttcaaca tgttctgggt gcgacaggcg 120 cccggcaagg ggctggaata cgtcgctcaa attagcagca gtggtagtag cacaaactac 180 gcacccgcgg tgaggggccg tgccaccatc tcgagggaca acgggcagag cacagtgagg 240 ctgcagctga acaaccccgg ggctgaagac accggcacct actactgcgc caaaagtagt 300 tatgactgtc cttacggtca ttgtagtagt ggtgttgata gtgctggtga gatcgacgca 360 tggggccacg ggaccgaagt catcgtctcc tcca 394 <210> 45 <211> 366 <212> DNA <213> Artificial sequence <220> <223> VH of antibody 1-65 <400> 45 gccgtgacgt tggacgagtc cgggggcggc ctccagacgc ccggaggagc gctcagcctc 6​​​​​​​​​​​​​​​​​​​​​​​​​​​gccgtgacgt tggacgagtc cgggggcggc ctccagacgc ccggaggagc gctcagcctc 60 gtctgcaagg cctccgggtt cgacttcagc gattatggca tgggttgggt gcgacaggct 120 ccaggcaagg ggctggagtg ggttgctgct attagaagtg atggtagtaa cccatcatac 180 gggtcggcgg tgaagggccg tgccaccatc tcgaaggaca acgggcgaag cacagtgagg 240 ctgcagctga acaacctcag ggctgaggac accgccacct actactgcgc caaggatggt 300 aatggttact gtgctctcga tgcttatcgt agtggtggtt atagttgtgg tgtttatcct 360 ggtagcatcg acgcatgggg ccacgggacc gaagtcatcg tctcctcc 408 <210> 47 <211> 315 <212> DNA <213> Artificial Sequence <220> <223> 2-13 Antibody VL <400> 47 ggccctgact cagccgtcct cggtgtcagc aaacccagga gaaaccgtca agataacctg 60 ctccgggggt agctatagct atggctggtt ccagcagaag tctcctggca gtgcccttgt 120 cactgtgatc tactgggatg atgagagacc ctcggacatc ccttcacgat tctccggtgc 180 cctatccggc tccacaaaca cattaaccat cactggggtc caagccgacg acgaggctgt 240 ctatttctgt gggactgaag acatcagcgg cactgctggt gtatttgggg ccgggacaac 300 cctgaccgtc ctggg 315 <210> 48 <211> 327 <212> DNA <213> Artificial sequence <220> <223> VL of 3-2 antibody <400> 48 ggccctgact cagccgtcct cggtgtcagc aaacccagga gaaaccgtca agatcacctg 60 ctccgggggt ggcagctatg ctggaagtta ctattatggc tggtaccagc agaaggcacc 120 tggcagtgcc cctgtcactc tgatctatga aagcaacaag agaccctcgg acatcccttc 180 acgattctcc ggttccacat ctggctccac agccacacta accatcactg gggtccaagc 240 cgatgacgag gctatctatt actgtgggag ctgggacagt agcaatggtg gtatatttgg 300 ggccgggaca accctgaccg tcctagg 327 <210> 49 <211> 331 <212> DNA <213> Artificial sequence <220> <223> VL of 1-65 antibody <400> 49 ggccctgact cagccgtcct cggtgtcagc aaaccctggg gaaactgtca agatcacctg 60 ctccgggggt ggtagcagtg gctatggtta tggctggtat cagcagaagt cacctagcag 120 tgcccctctc actgtgatct actggaacga caagagaccc tcggacatcc cttcacgatt 180 ctccggttcc aaatccggct ccacacacac attaaccatc actggggtcc aagccgagga 240 cgaggctgta tatttctgtg ggaatgatga ctacagcagt gatagtggat atgtcggtgt 300 atttggggcc gggacaaccc tgaccgtcct a 331 <210> 50 <211> 312 <212> DNA <213> Artificial Sequence <220> <223> 1-28 Antibody VL <400> 50 gccctgactc agccgtcctc ggtgtcagca aacctggaag gaaccgtcga gatcacctgc 60 tccgggagtg gctatggtta tggctggtat cagcagaagt ctcctggcag tgcccctgtc 120 actgtgatct atcagaacga caagagaccc tcggacatcc cttcacgatt ctccggttcc 180 aaatccggct ccacgggcac attaaccatc actggggtcc aagtcgagga cgaggctgtc 240 tattactgtg ggagtgaaga cagcagcact cttgctggta tatttggggc cgggacaacc 300 ctgaccgtcc ta 312 <210> 51 <211> 100 <212> PRT <213> Artificial sequence <220> <223> M3 mutant <400> 51 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Ile Glu Glu Arg Ser Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 52 <211> 100 <212> PRT <213> Artificial sequence <220> <223> M4 Mutant <400> 52 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Val Lys Cys Ser Cys Phe Pro Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 53 <211> 100 <212> PRT <213> Artificial Sequence <220> <223> M5 Mutant <400> 53 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Asn Lys Pro Ser Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 54 <211> 100 <212> PRT <213> Artificial Sequence <220> <223> M6 Mutant <400> 54 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Leu Gln Arg Trp Trp 50 55 60 Cys Gln Met Glu Leu Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 55 <211> 100 <212> PRT <213> Artificial Sequence <220> <223> M7 Mutant <400> 55 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Glu Cys Lys Thr Leu Pro 65 70 75 80 Asp Asn Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 56 <211> 101 <212> PRT <213> Artificial sequence <220> <223> M8 mutant <400> 56 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Ser Cys Ser Ser Gly Asn Lys Ile Lys 85 90 95 Thr Thr Thr Val Ser 100 <210> 57 <211> 459 <212> PRT <213> Artificial sequence <220> <223> 2-13 Antibody HC <400> 57 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30 Gly Met Phe Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu Tyr Val 35 40 45 Ala Glu Ile Thr Asn Asp Gly Ser Gly Thr Asn Tyr Gly Ser Ala Val 50 55 60 Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Gly Thr Tyr Phe Cys 85 90 95 Ala Arg Ser Thr Tyr Glu Cys Pro Gly Gly Phe Ser Cys Trp Gly Asp 100 105 110 Thr Gly Gln Ile Asp Ala Trp Gly His Gly Thr Glu Val Ile Val Ser 115 120 125 Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser 130 135 140 Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 145 150 155 160 Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr 165 170 175 Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr 180 185 190 Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln 195 200 205 Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp 210 215 220 Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro 225 230 235 240 Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 245 250 255 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 260 265 270 Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 275 280 285 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 290 295 300 Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 305 310 315 320 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 325 330 335 Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys 340 345 350 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 355 360 365 Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 370 375 380 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 385 390 395 400 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 405 410 415 Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 420 425 430 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 435 440 445 Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 <210> 58 <211> 461 <212> PRT <213> Artificial Sequence <220> <223> 3-2 antibody HC <400> 58 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30 Asn Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Val 35 40 45 Ala Gln Ile Ser Ser Ser Gly Ser Ser Thr Asn Tyr Ala Pro Ala Val 50 55 60 Arg Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Pro Gly Ala Glu Asp Thr Gly Thr Tyr Tyr Cys 85 90 95 Ala Lys Ser Ser Tyr Asp Cys Pro Tyr Gly His Cys Ser Ser Gly Val 100 105 110 Asp Ser Ala Gly Glu Ile Asp Ala Trp Gly His Gly Thr Glu Val Ile 115 120 125 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 130 135 140 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 145 150 155 160 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 165 170 175 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 180 185 190 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 195 200 205 Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 210 215 220 Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 225 230 235 240 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 245 250 255 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 260 265 270 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 275 280 285 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 290 295 300 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 305 310 315 320 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 325 330 335 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 340 345 350 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 355 360 365 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 370 375 380 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 385 390 395 400 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 405 410 415 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 420 425 430 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 435 440 445 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 460 <210> 59 <211> 452 <212> PRT <213> Artificial sequence <220> <223> 1-65 antibody HC <400> 59 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gln Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Val Ile Asn Lys Ser Gly Ser Asp Thr Ser Tyr Gly Ser Ala Val 50 55 60 Lys Gly Arg Ala Thr Ile Ser Arg Asp Asn Gly Gln Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Gly Thr Tyr Phe Cys 85 90 95 Ala Lys Gly Ser Ala Ser Tyr Ile Thr Ala Ala Thr Ile Asp Ala Trp 100 105 110 Gly His Gly Thr Glu Val Ile Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 60 <211> 466 <212> PRT <213> Artificial Sequence <220> <223> 1-28 Antibody HC <400> 60 Ala Val Thr Leu Asp Glu Ser Gly Gly Gly Leu Gln Thr Pro Gly Gly 1 5 10 15 Ala Leu Ser Leu Val Cys Lys Ala Ser Gly Phe Asp Phe Ser Asp Tyr 20 25 30 Gly Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Arg Ser Asp Gly Ser Asn Pro Ser Tyr Gly Ser Ala Val 50 55 60 Lys Gly Arg Ala Thr Ile Ser Lys Asp Asn Gly Arg Ser Thr Val Arg 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Lys Asp Gly Asn Gly Tyr Cys Ala Leu Asp Ala Tyr Arg Ser Gly 100 105 110 Gly Tyr Ser Cys Gly Val Tyr Pro Gly Ser Ile Asp Ala Trp Gly His 115 120 125 Gly Thr Glu Val Ile Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 130 135 140 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 145 150 155 160 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 165 170 175 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 180 185 190 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 195 200 205 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 210 215 220 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 225 230 235 240 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 245 250 255 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 260 265 270 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 275 280 285 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 290 295 300 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 305 310 315 320 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 325 330 335 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 340 345 350 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 355 360 365 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 370 375 380 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 385 390 395 400 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 405 410 415 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 420 425 430 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 435 440 445 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 450 455 460 Gly Lys 465 <210> 61 <211> 211 <212> PRT <213> Artificial Sequence <220> <223> 2 - 13 Antibody LC <400> 61 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu Thr Val 1 5 10 15 Lys Ile Thr Cys Ser Gly Gly Ser Tyr Ser Tyr Gly Trp Phe Gln Gln 20 25 30 Lys Ser Pro Gly Ser Ala Leu Val Thr Val Ile Tyr Trp Asp Asp Glu 35 40 45 Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ala Leu Ser Gly Ser 50 55 60 Thr Asn Thr Leu Thr Ile Thr Gly Val Gln Ala Asp Asp Glu Ala Val 65 70 75 80 Tyr Phe Cys Gly Thr Glu Asp Ile Ser Gly Thr Ala Gly Val Phe Gly 85 90 95 Ala Gly Thr Thr Leu Thr Val Leu Arg Ser Val Ala Ala Pro Ser Val 100 105 110 Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser 115 120 125 Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln 130 135 140 Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val 145 150 155 160 Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu 165 170 175 Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu 180 185 190 Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg 195 200 205 Gly Glu Cys 210 <210> 62 <211> 215 <212> PRT <213> artificial sequence <220> <223> 3-2 antibody LC <400> 62 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu Thr Val 1 5 10 15 Lys Ile Thr Cys Ser Gly Gly Gly Ser Tyr Ala Gly Ser Tyr Tyr Tyr 20 25 30 Gly Trp Tyr Gln Gln Lys Ala Pro Gly Ser Ala Pro Val Thr Leu Ile 35 40 45 Tyr Glu Ser Asn Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Thr Ser Gly Ser Thr Ala Thr Leu Thr Ile Thr Gly Val Gln Ala 65 70 75 80 Asp Asp Glu Ala Ile Tyr Tyr Cys Gly Ser Trp Asp Ser Ser Asn Gly 85 90 95 Gly Ile Phe Gly Ala Gly Thr Thr Leu Thr Val Leu Arg Ser Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 63 <211> 217 <212> PRT <213> Artificial Sequence <220> <223> 1-65 Antibody LC <400> 63 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Pro Gly Glu Thr Val 1 5 10 15 Lys Ile Thr Cys Ser Gly Gly Gly Ser Ser Gly Tyr Gly Tyr Gly Trp 20 25 30 Tyr Gln Gln Lys Ser Pro Ser Ser Ala Pro Leu Thr Val Ile Tyr Trp 35 40 45 Asn Asp Lys Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Lys 50 55 60 Ser Gly Ser Thr His Thr Leu Thr Ile Thr Gly Val Gln Ala Glu Asp 65 70 75 80 Glu Ala Val Tyr Phe Cys Gly Asn Asp Asp Tyr Ser Ser Asp Ser Gly 85 90 95 Tyr Val Gly Val Phe Gly Ala Gly Thr Thr Leu Thr Val Leu Arg Ser 100 105 110 Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu 115 120 125 Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro 130 135 140 Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly 145 150 155 160 Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His 180 185 190 Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val 195 200 205 Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 64 <211> 211 <212> PRT <213> Synthetic Sequence <220>[[ID=3f]] <223> 1-28 Antibody LC <400> 64 Ala Leu Thr Gln Pro Ser Ser Val Ser Ala Asn Leu Glu Gly Thr Val ID=44]]1 5 10 15 Glu Ile Thr Cys Ser Gly Ser Gly Tyr Gly Tyr Gly Trp Tyr Gln Gln 20 25 30 Lys Ser Pro Gly Ser Ala Pro Val Thr Val Ile Tyr Gln Asn Asp Lys 35 40 45 Arg Pro Ser Asp Ile Pro Ser Arg Phe Ser Gly Ser Lys Ser Gly Ser 50 55 60 Thr Gly Thr Leu Thr Ile Thr Gly Val Gln Val Glu Asp Glu Ala Val 65 70 75 80 Tyr Tyr Cys Gly Ser Glu Asp Ser Ser Thr Leu Ala Gly Ile Phe Gly 85 90 95 Ala Gly Thr Thr Leu Thr Val Leu Arg Ser Val Ala Ala Pro Ser Val 100 105 110 Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser 115 120 125 Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln 130 135 140 Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val 145 150 155 160 Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu 165 170 175 Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu 180 185 190 Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg 195 200 205 Gly Glu Cys 210 <210> 65 <211> 5 <212> PRT <213> Artificial sequence <220> <223> EP1 epitope <400> 65 Ile Val Thr Leu Asp 1 5 <210> 66 <211> 5 <212> PRT <213> Artificial sequence <220> <223> EP2 epitope <400> 66 Asp Ser Ser Gln Pro 1 5 <210> 67 <211> 5 <212> PRT <213> Artificial sequence <220> <223> EP3 epitope <400> 67 Arg Thr Ile Ala Arg 1 5 <210> 68 <211> 7 <212> PRT <213> Artificial sequence <220> <223> EP4 epitope <400> 68 Ala Arg Cys Ala Cys Arg Lys 1 5 <210> 69 <211> 4 <212> PRT <213> Artificial sequence <220> <223> EP5 epitope <400> 69 Ala Arg Pro Ala 1 <210> 70 <211> 9 <212> PRT <213> Artificial sequence <220> <223> EP6 epitope <400> 70 Lys Thr Lys Gln Trp Cys Asp Met Leu 1 5 <210> 71 <211> 8 <212> PRT <213> Artificial sequence <220> <223> EP7 epitope <400> 71 Gly Cys Asp Leu Leu Ile Asn Arg 1 5 <210> 72 <211> 8 <212> PRT <213> Artificial sequence <220> <223> EP8 epitope <400> 72 Thr Cys Thr Gln Pro Gly Gly Arg 1 5 <210> 73 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F2-01-BSA (#1) <400> 73 Thr Ala Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 74 <211> 20 <212> PRT <213> Synthetic Sequence <220> <223> F2-02-BSA (#2) <400> 74 Thr Leu Ala Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Thr Ala Arg Cys 20 <210> 77 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F2-05-BSA (#5) <400> 77 Thr Leu Asp Arg Asp Ser Ala Gln Pro Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 [[ID=)22]]Thr Ala Arg Cys 20 <210> 78 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F2-06-BSA (#6) <400> 78 Thr Leu Asp Arg Asp Ser Ser Ala Pro Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 79 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F2-07-BSA (#7) <400> 79 Thr Leu Asp Arg Asp Ser Ser Gln Ala Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 80 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> F2-08-BSA (#8) <400> 80 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Ala Thr Ile Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 81 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> F2-09-BSA (#9) <400> 81 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Ala Ile Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 82 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> F2-10-BSA (#10) <400> 82 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ala Ala Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 83 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> F2-11-BSA (#11) <400> 83 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Arg Arg Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 84 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F2-12-BSA (#12) <400> 84 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Ala Gln 1 5 10 15 Thr Ala Arg Cys 20 <210> 85 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F2-12-BSA (#13) <400> 85 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 1 5 10 15 Thr Val Arg Cys 20 <210> 86 <211> 100 <212> PRT <213> Artificial sequence <220> <223> Wild-type FAM19A5 isoform 2 (without signal peptide) <400> 86 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Asp Ser Ser Gln Pro Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95<​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 88 <211> 100 <212> PRT <213> Artificial Sequence <220> <223> M2 Mutant <400> 88 Gln Phe Leu Lys Glu Gly Gln Leu Ala Ala Gly Thr Cys Glu Ile Val 1 5 10 15 Thr Leu Asp Arg Cys Cys Asn Lys Asn Arg Arg Thr Ile Ala Arg Gln 20 25 30 Thr Ala Arg Cys Ala Cys Arg Lys Gly Gln Ile Ala Gly Thr Thr Arg 35 40 45 Ala Arg Pro Ala Cys Val Asp Ala Arg Ile Ile Lys Thr Lys Gln Trp 50 55 60 Cys Asp Met Leu Pro Cys Leu Glu Gly Glu Gly Cys Asp Leu Leu Ile 65 70 75 80 Asn Arg Ser Gly Trp Thr Cys Thr Gln Pro Gly Gly Arg Ile Lys Thr 85 90 95 Thr Thr Val Ser 100 <210> 89 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P2-C12 antibody VH CDR1 <400> 89 Thr Tyr Ala Val Thr 1 5 <210> 90 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P2-C12 antibody VH CDR2 <400> 90 Tyr Ile Asn Trp Arg Gly Gly Thr Ser Tyr Ala Asn Trp Ala Lys Gly 1 5 10 15 <210> 91 <211> 15 <212> PRT <213> Artificial sequence <220> <223> P2-C12 antibody VH CDR3 <400> 91 Asp Ala Ser Ser Gly Ala Ala Phe Gly Ser Tyr Gly Met Asp Pro 1 5 10 15 <210> 92 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P2-C12 Antibody VL CDR1 <400> 92 Gln Ala Ser Gln Ser Ile Ser Ser Tyr Leu Ser 1 5 10 <210> 93 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P2-C12 antibody VL CDR2 <400> 93 Glu Ala Ser Lys Leu Ala Ser 1 5 <210> 94 <211> 12 <212> PRT <213> Artificial sequence <220> <223> P2-C12 Antibody VL CDR3 <400> 94 Gln Gln Gly Tyr Ser Ser Thr Asn Val Trp Asn Ala 1 5 10 <210> 95 <211> 6 <212> PRT <213> Artificial sequence <220> <223> 13B4 antibody VH CDR1 <400> 95 Ser Ser Asn Trp Trp Ser 1 5 <210> 96 <211> 16 <212> PRT <213> Artificial sequence <220> <223> 13B4 antibody VH CDR2 <400> 96 Glu Ile Tyr His Gly Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys Gly 1 5 10 15 <210> 97 <211> 9 <212> PRT <213> Artificial sequence <220> <223> 13B4 antibody VH CDR3 <400> 97 Trp Gln Leu Val Gly Gly Leu Asp Val 1 5 <210> 98 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 13B4 antibody VL CDR1 <400> 98 Ser Gly Asp Lys Leu Gly Asn Val Tyr Ala Ser 1 5 10 <210> 99 <211> 7 <212> PRT <213> Artificial sequence <220> <223> 13B4 antibody VL CDR2 <400> 99 Gln Asp Asn Lys Arg Pro Ser 1 5 <210> 100 <211> 9 <212> PRT <213> Artificial sequence <220> <223> 13B4 antibody VL CDR3 <400> 100 Gln Ala Trp Asp Ser Ser Thr Ala Val 1 5 <210> 101 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 13F7 antibody VH CDR1 <400> 101 Gly Tyr Ser Trp Thr 1 5 <210> 102 <211> 16 <212> PRT <213> Artificial sequence <220> <223> 13F7 antibody VH CDR2 <400> 102 Glu Ile Ser His Phe Gly Ser Ala Asn Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 103 <211> 15 <212> PRT <213> Artificial sequence <220> <223> 13F7 antibody VH CDR3 <400> 103 Ala Leu Arg Gly Thr Tyr Ser Arg Phe Tyr Tyr Gly Met Asp Val 1 5 10 15 <210> 104 <211> 16 <212> PRT <213> Artificial sequence <220> <223> 13F7 antibody VL CDR1 <400> 104 Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp 1 5 10 15 <210> 105 <211> 7 <212> PRT <213> Artificial sequence <220> <223> 13F7 antibody VL CDR2 <400> 105 Leu Gly Ser Asn Arg Ala Ser 1 5 <210> 106 <211> 9 <212> PRT <213> Artificial sequence <220> <223> 13F7 antibody VL CDR3 <400> 106 Met Gln Ala Arg Gln Thr Pro Leu Thr 1 5 <210> 107 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 15A9 antibody VH CDR1 <400> 107 Ser Tyr Tyr Trp Ser 1 5 <210> 108 <211> 16 <212> PRT <213> Artificial sequence <220> <223> 15A9 antibody VH CDR2 <400> 108 Tyr Ile Tyr Pro Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 109 <211> 12 <212> PRT <213> Artificial sequence <220> <223> 15A9 antibody VH CDR3 <400> 109 Val Asn Pro Phe Gly Tyr Tyr Tyr Ala Met Asp Val 1 5 10 <210> 110 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 15A9 antibody VL CDR1 <400> 110 Arg Ala Ser Gln Ser Ile Ser Thr Ser Leu Asn 1 5 10 <210> 111 <211> 7 <212> PRT <213> Artificial sequence <220> <223> 15A9 antibody VL CDR2 <400> 111 Gly Ala Ser Thr Leu Gln Ser 1 5 <210> 112 <211> 9 <212> PRT <213> Artificial sequence <220> <223> 15A9 antibody VL CDR3 <400> 112 Gln Glu Ser Ala Ser Ile Pro Arg Thr 1 5 <210> 113 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P1-A03 antibody VH CDR1 <400> 113 Ser Asp Tyr Met Ser 1 5 <210> 114 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P1-A03 antibody VH CDR2 <400> 114 Ile Ile Tyr Pro Ser Thr Thr Thr Tyr Tyr Tyr Ala Ser Trp Ala Lys Gly 1 5 10 15 <210> 115 <211> 10 <212> PRT <213> Artificial sequence <220> <223> P1-A03 antibody VH CDR3 <400> 115 Gly Ser Asn Trp Ser Ser Gly Met Asn Leu 1 5 10 <210> 116 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P1-A03 antibody VL CDR1 <400> 116 Leu Ala Ser Glu Asp Ile Tyr Ser Gly Ile Ser 1 5 10 <210> 117 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P1-A03 antibody VL CDR2 <400> 117 Gly Ala Ser Asn Leu Glu Ser 1 5 <210> 118 <211> 12 <212> PRT <213> Artificial sequence <220> <223> P1-A03 antibody VL CDR3 <400> 118 Leu Gly Gly Tyr Ser Tyr Ser Ser Thr Gly Leu Thr 1 5 10 <210> 119 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P1-A08 antibody VH CDR1 <400> 119 Thr Tyr Tyr Met Ser 1 5 <210> 120 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P1-A08 antibody VH CDR2 <400> 120 Ile Val Tyr Pro Ser Gly Thr Thr Tyr Tyr Ala Asn Trp Ala Lys Gly 1 5 10 15 <210> 121 <211> 8 <212> PRT <213> Artificial sequence <220> <223> P1-A08 antibody VH CDR3 <400> 121 Gly Asp Ser Phe Gly Tyr Gly Leu 1 5 <210> 122 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P1-A08 antibody VL CDR1 <400> 122 Thr Ala Asp Thr Leu Ser Arg Ser Tyr Ala Ser 1 5 10 <210> 123 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P1-A08 antibody VL CDR2 <400> 123 Arg Asp Thr Ser Arg Pro Ser 1 5 <210> 124 <211> 13 <212> PRT <213> Artificial sequence <220> <223> P1-A08 antibody VL CDR3 <400> 124 Ala Thr Ser Asp Gly Ser Gly Ser Asn Tyr Gln Tyr Val 1 5 10 <210> 125 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P1-F02 antibody VH CDR1 <400> 125 Asn Tyr Tyr Met Gly 1 5 <210> 126 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P1-F02 antibody VH CDR2 <400> 126 Ile Ile Tyr Ala Ser Gly Ser Thr Tyr Tyr Tyr Ala Ser Trp Ala Lys Gly 1 5 10 15 <210> 127 <211> 17 <212> PRT <213> Artificial sequence <220> <223> P1-F02 antibody VH CDR3 <400> 127 Ile Asp Ile Gly Val Gly Asp Tyr Gly Trp Ala Tyr Asp Arg Leu Asp 1 5 10 15 Leu <210> 128 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P1-F02 antibody VL CDR1 <400> 128 Leu Ala Ser Glu Asp Ile Tyr Ser Gly Ile Ser 1 5 10 <210> 129 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P1-F02 antibody VL CDR2 <400> 129 Gly Ala Ser Asn Leu Glu Ser 1 5 <210> 130 <211> 10 <212> PRT <213> Artificial sequence <220> <223> P1-F02 antibody VL CDR3 <400> 130 Leu Gly Gly Tyr Ser Tyr Ser Ser Ile Thr 1 5 10 <210> 131 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P2-A01 antibody VH CDR1 <400> 131 Gly Tyr Tyr Met Ser 1 5 <210> 132 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P2-A01 antibody VH CDR2 <400> 132 Ile Ile Tyr Pro Ser Gly Ser Thr Asp Tyr Ala Ser Trp Ala Lys Gly 1 5 10 15 <210> 133 <211> 15 <212> PRT <213> Artificial sequence <220> <223> P2-A01 antibody VH CDR3 <400> 133 Val Ala Gly Tyr Val Gly Tyr Gly Tyr Glu Thr Phe Phe Asp Ile 1 5 10 15 <210> 134 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P2-A01 Antibody VL CDR1 <400> 134 Leu Ala Ser Glu Asp Ile Tyr Ser Gly Ile Ser 1 5 10 <210> 135 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P2-A01 Antibody VL CDR2 <400> 135 Gly Ala Ser Asn Leu Glu Ser 1 5 <210> 136 <211> 14 <212> PRT <213> Artificial sequence <220> <223> P2-A01 Antibody VL CDR3 <400> 136 Leu Gly Gly Val Thr Tyr Ser Ser Thr Gly Thr His Leu Thr 1 5 10 <210> 137 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P2-A03 antibody VH CDR1 <400> 137 Asn Tyr Asp Met Ser 1 5 <210> 138 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P2-A03 antibody VH CDR2 <400> 138 Phe Met Asp Thr Asp Gly Ser Ala Tyr Tyr Ala Thr Trp Ala Lys Gly 1 5 10 15 <210> 139 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P2-A03 antibody VH CDR3 <400> 139 Arg Gly Ser Ser Tyr Tyr Gly Gly Ile Asp Ile 1 5 10 <210> 140 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P2-A03 antibody VL CDR1 <400> 140 Gln Ala Ser Gln Ser Ile Gly Gly Asn Leu Ala 1 5 10 <210> 141 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P2-A03 antibody VL CDR2 <400> 141 Arg Ala Ser Thr Leu Ala Ser 1 5 <210> 142 <211> 14 <212> PRT <213> Artificial sequence <220> <223> P2-A03 antibody VL CDR3 <400> 142 Gln Ser Pro Ala Tyr Asp Pro Ala Ala Tyr Val Gly Asn Ala 1 5 10 <210> 143 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P2-F07 antibody VH CDR1 <400> 143 Ser Tyr Tyr Met Asn 1 5 <210> 144 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P2-F07 antibody VH CDR2 <400> 144 Ile Ile Tyr Pro Ser Gly Thr Thr Tyr Tyr Tyr Ala Gly Trp Ala Lys Gly 1 5 10 15 <210> 145 <211> 8 <212> PRT <213> Artificial sequence <220> <223> P2-F07 antibody VH CDR3 <400> 145 Thr Val Ser Gly Tyr Phe Asp Ile 1 5 <210> 146 <211> 11 <212> PRT <213> Artificial sequence <220> <223> P2-F07 antibody VL CDR1 <400> 146 Leu Ala Ser Glu Asp Ile Tyr Ser Ala Leu Ala 1 5 10 <210> 147 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P2-F07 antibody VL CDR2 <400> 147 Gly Thr Ser Asn Leu Glu Ser 1 5 <210> 148 <211> 9 <212> PRT <213> Artificial sequence <220> <223> P2-F07 antibody VL CDR3 <400> 148 Gln Gly Tyr Ser Ser Tyr Pro Leu Thr 1 5 <210> 149 <211> 5 <212> PRT <213> Artificial sequence <220> <223> P2-F11 antibody VH CDR1 <400> 149 Ser Tyr Gly Val Ser 1 5 <210> 150 <211> 16 <212> PRT <213> Artificial sequence <220> <223> P2-F11 antibody VH CDR2 <400> 150 Tyr Ile Ala Asn Asn Tyr Asn Pro His Tyr Ala Ser Trp Ala Lys Gly 1 5 10 15 <210> 151 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P2-F11 antibody VH CDR3 <400> 151 Asp Asn Tyr Gly Met Asp Pro 1 5 <210> 152 <211> 13 <212> PRT <213> Artificial sequence <220> <223> P2-F11 antibody VL CDR1 <400> 152 Gln Ala Ser Gln Ser Val Tyr Asn Asn Lys Asn Leu Ala 1 5 10 <210> 153 <211> 7 <212> PRT <213> Artificial sequence <220> <223> P2-F11 antibody VL CDR2 <400> 153 Ala Ala Ser Thr Leu Ala Ser 1 5 <210> 154 <211> 13 <212> PRT <213> Artificial sequence <220> <223> P2-F11 antibody VL CDR3 <400> 154 Gln Gly Glu Phe Ser Cys Ser Ser Ala Asp Cys Asn Ala 1 5 10 <210> 155 <211> 121 <212> PRT <213> Artificial sequence <220> <223> P2-C12 antibody VH <400> 155 Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Thr Tyr Ala 20 25 30 Val Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly 35 40 45 Tyr Ile Asn Trp Arg Gly Gly Thr Ser Tyr Ala Asn Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys Met 65 70 75 80 Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp 85 90 95 Ala Ser Ser Gly Ala Ala Phe Gly Ser Tyr Gly Met Asp Pro Trp Gly 100 105 110 Pro Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 156 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> 13B4 antibody VH <400> 156 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly 1 5 10 15 Thr Leu Ser Leu Asn Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser 20 25 30 Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Glu Ile Tyr His Gly Gly Thr Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Gly Arg Val Thr Met Ser Val Asp Lys Thr Lys Asn Gln Phe Ser 65 70 75 80 Leu Arg Leu Ser Ser Val Thr Ala Val Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Gln Leu Val Gly Gly Leu Asp Val Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser 115 <210> 157 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> VH of 13F7 Antibody <400> 157 Gln Val Gln Leu Gln Glu Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Asn Ala Glu Ser Phe Asn Gly Tyr 20 25 30 Ser Trp Thr Trp Ile Arg Gln Thr Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Ser His Phe Gly Ser Ala Asn Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Ala Thr Ile Ser Ala Asp Lys Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Lys Leu Thr Ser Val Thr Ala Val Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Leu Arg Gly Thr Tyr Ser Arg Phe Tyr Tyr Gly Met Asp Val 100 105 110 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 158 <211> 120 <212> PRT <213> artificial sequence <220> <223> 15A9 antibodyVH <400> 158 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Tyr 20 25 30 Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Tyr Pro Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Asn Leu Lys Ser Val Thr Ala Val Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Val Asn Pro Phe Gly Tyr Tyr Tyr Ala Met Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 159 <211> 116 <212> PRT <213> Synthetic Sequence <220> <223> VH of P1 - A03 Antibody <400> 159 Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Asp Tyr 20 25 30 Met Ser Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Glu Trp Ile Gly 35 40 45 Ile Ile Tyr Pro Ser Thr Thr Thr Tyr Tyr Ala Ser Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Glu Leu Lys Met 65 70 75 80 Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly 85 90 95 Ser Asn Trp Ser Ser Gly Met Asn Leu Trp Gly Pro Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <210> 160 <211> 113 <212> PRT <213> Artificial sequence <220> <223> VH of P1-A08 antibody <400> 160 Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Thr Tyr Tyr 20 25 30 Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly 35 40 45 Ile Val Tyr Pro Ser Gly Thr Thr Tyr Tyr Ala Asn Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Thr Ala Ser Thr Thr Val Asp Leu Met Ile Thr 65 70 75 80 Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Asp 85 90 95 Ser Phe Gly Tyr Gly Leu Trp Gly Pro Gly Thr Leu Val Thr Val Ser 100 105 110 Ser <210> 161 <211> 122 <212> PRT <213> Artificial sequence <220> <223> VH of P1-F02 antibody <400> 161 Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Asn Tyr Tyr 20 25 30<​​​​​​​​​​​​​​​​​​​​​​​​​​​​<212> PRT <213> Artificial sequence <220> <223> VH of P2 - A01 antibody <400> 162 Gln Glu Gln Leu Val Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr 1 5 10 15 Pro Leu Thr Leu Ser Cys Thr Ala Ser Gly Phe Phe Leu Ser Gly Tyr 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ile Ile Tyr Pro Ser Gly Ser Thr Asp Tyr Ala Ser Trp Ala Lys 50 55 60 Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile 65 70 75 80 Thr Thr Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Val 85 90 95 Ala Gly Tyr Val Gly Tyr Gly Tyr Glu Thr Phe Phe Asp Ile Trp Gly 100 105 110 Pro Gly Thr Leu Val Thr Val Ser Leu 115 120 <210> 163 <211> 116 <212> PRT <213> Artificial sequence <220> <223> VH of P2 - A03 antibody <400> 163 Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Asn Asn Tyr Asp 20 25 30 Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly 35 40 45 Phe Met Asp Thr Asp Gly Ser Ala Tyr Tyr Ala Thr Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Val Asp Leu Lys Met Thr 65 70 75 80 Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Arg Gly 85 90 95 Ser Ser Tyr Tyr Gly Gly Ile Asp Ile Trp Gly Pro Gly Thr Pro Val 100 105 110 Thr Val Ser Leu 115 <210> 164 <211> 113 <212> PRT<00…​​​​​​​​​​1 5 10 15 Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Ser Tyr Tyr 20 25 30 Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly 35 40 45 Ile Ile Tyr Pro Ser Gly Thr Thr Tyr Tyr Ala Gly Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr 65 70 75 80 Ser Pro Thr Ser Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Thr Val 85 90 95 Ser Gly Tyr Phe Asp Ile Trp Gly Pro Gly Thr Leu Val Thr Val Ser 100 105 110 Leu <210> 165 <211> 114 <212> PRT <213> Synthetic Sequence <220> <223> VH of P2-F11 antibody <400> 165 Gln Glu Gln Leu Val Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr 20 25 30 Gly Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Ala Asn Asn Tyr Asn Pro His Tyr Ala Ser Trp Ala Lys 50 55 60 Gly Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys 65 70 75 80 Met Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg 85 90 95 Asp Asn Tyr Gly Met Asp Pro Trp Gly Pro Gly Thr Leu Val Thr Val 100 105 110 Ser Ser <210> 166 <211> 110 <212> PRT <213> Synthetic sequence <220> <223> P2-C12 antibody VL <400> 166 Glu Leu Asp Met Thr Gln Thr Pro Ser Ser Val Ser Ala Ala Val Gly 1 5 10 15 Gly Thr Val Thr Ile Lys Cys Gln Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Glu Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Glu Phe Thr Leu Thr Ile Ser Asp Leu Glu Cys 65 70 75 80 Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Ser Ser Thr Asn 85 90 95 Val Trp Asn Ala Phe Gly Gly Gly Thr Asn Val Glu Ile Lys 100 105 110 <210> 167 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> 13B4 antibody VL <400> 167 Ser Tyr Glu Leu Thr Gln Pro Leu Ser Val Ser Val Ser Pro Gly Gln 1 5 10 15 Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asn Val Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Thr Leu Val Ile Tyr 35 40 45 Gln Asp Asn Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Lys Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Leu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser Thr Ala Val 85 90 95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 <210> 168 <211> 112 <212> PRT <213> Artificial Sequence <220> <223> VL of 13F7 Antibody <400> 168 Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ala Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20 25 30 Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Val Gln Lys Pro Gly Gln Pro 35 40 45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala 85 90 95 Arg Gln Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 169 <211> 107 <212> PRT <213> Synthetic Sequence <220> <223> 15A9 Antibody VL <400> 169 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Ile Thr Ile Ser Cys Arg Ala Ser Gln Ser Ile Ser Thr Ser 20 25 30 Leu Asn Trp Tyr Gln Gln Thr Pro Gly Lys Ala Pro Arg Leu Leu Ile 35 40 45[[ID=二十九]] 50 55 60 Gly Gly Ser Gly Thr Asp Phe Ser Leu Thr Ile Thr Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Glu Ser Ala Ser Ile Pro Arg 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Asp Ile Lys 100 105 <210> 170 <211> 110 <212> PRT <213> Synthetic Sequence​ <220> <223> P1-A03 antibody VL <400> 170 Glu Leu Val Met Thr Gln Thr Pro Pro Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Thr Val Arg Ile Arg Cys Leu Ala Ser Glu Asp Ile Tyr Ser Gly 20 25 30 Ile Ser Trp Tyr Gln Gln Lys Pro Glu Lys Pro Pro Thr Leu Leu Ile 35 40 45 Ser Gly Ala Ser Asn Leu Glu Ser Gly Val Pro Pro Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Gly Gly Val Gln Ala 65 70 75 80 Glu Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Gly Tyr Ser Tyr Ser Ser 85 90 95 Thr Gly Leu Thr Phe Gly Ala Gly Thr Asn Val Glu Ile Lys 100 105 110 <210> 171 <211> 110 <212> PRT <213> artificial sequence <220> <223> P1-A08 antibody VL <400> 171 Glu Leu Val Leu Thr Gln Ser Pro Ser Val Gln Val Asn Leu Gly Gln 1 5 10 15 Thr Val Ser Leu Thr Cys Thr Ala Asp Thr Leu Ser Arg Ser Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Leu Ile Tyr 35 40 45 Arg Asp Thr Ser Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Ala Gln Ala Gly 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Ser Asp Gly Ser Gly Ser Asn 85 90 95 Tyr Gln Tyr Val Phe Gly Gly Gly Thr Gln Leu Thr Val Thr 100 105 110 <210> 172 <211> 108 <212> PRT <213> artificial sequence <220> <223> P1-F02 antibody VL <400> 172 Glu Leu Asp Met Thr Gln Thr Pro Pro Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Thr Val Arg Ile Arg Cys Leu Ala Ser Glu Asp Ile Tyr Ser Gly 20 25 30 Ile Ser Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro Thr Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Leu Glu Ser Gly Val Pro Pro Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Gly Gly Val Gln Ala 65 70 75 80 Glu Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Gly Tyr Ser Tyr Ser Ser 85 90 95 Ile Thr Phe Gly Ala Gly Thr Asn Val Glu Ile Lys 100 105 <210> 173 <211> 112 <212> PRT <213> artificial sequence <220> <223> P2-A01 antibody VL <400> 173 Glu Leu Val Met Thr Gln Thr Pro Pro Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Thr Val Arg Ile Arg Cys Leu Ala Ser Glu Asp Ile Tyr Ser Gly 20 25 30 Ile Ser Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro Thr Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Leu Glu Ser Gly Val Pro Pro Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Asp Tyr Thr Leu Thr Ile Gly Gly Val Gln Ala 65 70 75 80 Glu Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Gly Val Thr Tyr Ser Ser 85 90 95 Thr Gly Thr His Leu Thr Phe Gly Ala Gly Thr Asn Val Glu Ile Lys 100 105 110 <210> 174 <211> 112 <212> PRT <213> Artificial Sequence <220> <223> VL of P2 - A03 Antibody <400> 174 Glu Leu Asp Leu Thr Gln Thr Pro Ala Ser Val Ser Glu Pro Val Gly 1 5 10 15 Gly Thr Val Thr Ile Lys Cys Gln Ala Ser Gln Ser Ile Gly Gly Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Arg Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Lys Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asp Leu Glu Cys 65 70 75 80 Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Ser Pro Ala Tyr Asp Pro Ala 85 90 95 Ala Tyr Val Gly Asn Ala Phe Gly Gly Gly Thr Glu Leu Glu Ile Leu 100 105 110 <210> 175 <211> 107 <212> PRT <213> Synthetic Sequence <220> <223> VL of P2-F07 Antibody <400> 175 Glu Leu Asp Leu Thr Gln Thr Pro Pro Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Gly Thr Val Thr Ile Asn Cys Leu Ala Ser Glu Asp Ile Tyr Ser Ala 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro Thr Leu Leu Ile 35 40 45 Ser Gly Thr Ser Asn Leu Glu Ser Gly Val Pro Pro Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Gly Gly Val Gln Ala 65 70 75 80 Glu Asp Ala Ala Thr Tyr Phe Cys Gln Gly Tyr Ser Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Asn Val Glu Ile Lys 100 105 <210> 176 <211> 113 <212> PRT <213> Artificial sequence <220> <223> VL of P2-F11 antibody <400> 176 Glu Leu Asp Leu Thr Gln Thr Pro Ser Ser Val Ser Ala Ala Val Gly 1 5 10 15 Gly Thr Val Thr Ile Asn Cys Gln Ala Ser Gln Ser Val Tyr Asn Asn 20 25 30 Lys Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu 35 40 45 Leu Ile Tyr Ala Ala Ser Thr Leu Ala Ser Gly Val Ser Ser Arg Phe 50 55 60 Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val 65 70 75 80 Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Glu Phe Ser Cys 85 90 95 Ser Ser Ala Asp Cys Asn Ala Phe Gly Gly Gly Thr Glu Leu Glu Ile 100 105 110 Leu <210> 177 <211> 363 <212> DNA <213> Artificial sequence <220> <223> P2-C12VH <400> 177 cagtcgctgg aggagtccgg gggtcgcctg gtcacgcctg ggacacccct gacactcacc 120. tgcaccgtct ctggattctc cctcagtacc tatgcagtga cctgggtccg ccaggctcca gggaaggggc tggaatggat cggatacatt aattggcgtg gtgggacatc ctacgcgaac 180 240. tgggcgaaag gccgattcac catctccaaa acctcgtcga ccacggtgga tctgaaaatg accagtccga caaccgagga cacggccacc tatttctgtg ccagagatgc tagtagtggt gctgcttttg ggtcttacgg catggacccc tggggcccag ggaccctcgt caccgtctct 360 tca 363 <210> 178 <211> 330 <212> DNA <213> The snowstorm <220> <223> P2-C12VL <400> 178 60. gagctcgata tgacccagac tccatcctcc gtgtctgcag ctgtgggagg cacagtcacc atcaagtgcc aggccagtca gagcattagt agctacttat cctggtatca gcagaaacca gggcagcctc ccaagctcct gatctatgaa gcatccaaac tggcctctgg ggtcccatcg cggttcagcg gcagtggata tgggacagag ttcactctca ccatcagcga cctggagtgt 240 gccgatgctg ccacttacta ctgtcaacag ggttatagta gtactaatgt ttggaatgct 300 ttcggcggag gcaccaatgt ggaaatcaaa 330 <210> 179 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> P2-C12 HC <400> 179 Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Thr Tyr Ala 20 25 30 Val Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly 35 40 45 Tyr Ile Asn Trp Arg Gly Gly Thr Ser Tyr Ala Asn Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys Met 65 70 75 80 Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp<​​Ala Ser Ser Gly Ala Ala Phe Gly Ser Tyr Gly Met Asp Pro Trp Gly 100 105 110 Pro Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 180 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> 13B4 HC <400> 180 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly 1 5 10 15 Thr Leu Ser Leu Asn Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser 20 25 30 Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Glu Ile Tyr His Gly Gly Thr Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Gly Arg Val Thr Met Ser Val Asp Lys Thr Lys Asn Gln Phe Ser 65 70 75 80 Leu Arg Leu Ser Ser Val Thr Ala Val Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Gln Leu Val Gly Gly Leu Asp Val Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 181 <211> 453 <212> PRT <213> Artificial sequence <220> <223> 13F7 HC <400> 181 Gln Val Gln Leu Gln Glu Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Asn Ala Glu Ser Phe Asn Gly Tyr 20 25 � Ser Trp Thr Trp Ile Arg Gln Thr Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Ser His Phe Gly Ser Ala Asn Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Ala Thr Ile Ser Ala Asp Lys Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Lys Leu Thr Ser Val Thr Ala Val Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Leu Arg Gly Thr Tyr Ser Arg Phe Tyr Tyr Gly Met Asp Val 100 105 110 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln 355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450 <210> 182 <211> 450 <212> PRT <213> Artificial Sequence <220> <223> 15A9 HC <400> 182 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Tyr 20 25 30 Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Tyr Pro Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Asn Leu Lys Ser Val Thr Ala Val Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Val Asn Pro Phe Gly Tyr Tyr Tyr Ala Met Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210 > 183 <211 > 446 <212 > PRT <213 > Artificial sequence <220> <223 > P1 - A03 HC <400 > 183 Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Asp Tyr 20 25 30 Met Ser Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Glu Trp Ile Gly 35 40 45 Ile Ile Tyr Pro Ser Thr Thr Thr Tyr Tyr Ala Ser Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Glu Leu Lys Met 65 70 75 80 Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly 85 90 95 Ser Asn Trp Ser Ser Gly Met Asn Leu Trp Gly Pro Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 184 <211> 443 <212> PRT <213> Artificial Sequence <220> <223> P1-A08 HC <400> 184 Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 1 5 10 15 Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Thr Tyr Tyr 20 25 30 Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly 35 40 45 Ile Val Tyr Pro Ser Gly Thr Thr Tyr Tyr Ala Asn Trp Ala Lys Gly 50 55 60 Arg Phe Thr Ile Ser Thr Ala Ser Thr Thr Val Asp Leu Met Ile Thr 65 70 75 80 Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Asp 85 90 95 Ser Phe Gly Tyr Gly Leu Trp Gly Pro Gly Thr Leu Val Thr Val Ser 100 105 110 Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser 115 120 125 Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 130 135 140 Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr 145 150 155 160 Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr 165 170 175 Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln 180 185 190 Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp 195 200 205 Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro 210 215 220 Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 225 230 235 240 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 245 250 255 Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 260 265 270 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 275 280 285 Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 290 295 300 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 305 310 315 320 Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys 325 330 335 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 340 345 350 Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 355 360 365 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 370 375 380 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 385 390 395 400 Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 405 410 415 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 420 425 430 Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440...

Claims

1. Use of an antagonist to a family of sequence similarity 19, member A5 (FAM19A5) protein ("FAM19A5 antagonist") for the preparation of a medicament for treating neuropathic pain in a subject in need thereof, wherein the FAM19A5 antagonist is an anti-FAM19A5 antibody or an antigen-binding fragment thereof, or a polynucleotide encoding an anti-FAM19A5 antibody, wherein the anti-FAM19A5 antibody comprises a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, wherein: (a) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 214, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 217, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (b) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 15, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 27, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 28; (c) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:207, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:208, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:213, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:28; (d) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 214, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 215, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (e) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 218, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (f) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:217, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:216; (g) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:219, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:220; or (h) the heavy chain CDR1 consists of the amino acid sequence shown in SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence shown in SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence shown in SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence shown in SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence shown in SEQ ID NO: 221, and the light chain CDR3 consists of the amino acid sequence shown in SEQ ID NO:

222.

2. The use according to claim 1, wherein the neuropathic pain is central neuropathic pain or peripheral neuropathic pain.

3. The use according to claim 1, wherein the neuropathic pain is associated with physical injury, infection, diabetes, cancer therapy, alcoholism, amputation, muscle weakness in the back, legs, buttocks or face, trigeminal neuralgia, multiple sclerosis, herpes zoster, spinal surgery or any combination thereof.

4. The method of claim 1, wherein the neuropathic pain comprises carpal tunnel syndrome, central pain syndrome, degenerative disc disease, diabetic neuropathy, phantom limb pain, postherpetic neuralgia (shingles), pudendal neuralgia, sciatica, low back pain, trigeminal neuralgia, or any combination thereof.

5. The use according to claim 4, wherein the diabetic neuropathy is diabetic peripheral neuropathy.

6. The use according to claim 4, wherein the neuropathic pain is sciatica.

7. Use of a FAM19A5 antagonist for preparing a medicament for increasing the threshold or latency of a subject in need thereof to an external stimulus, wherein the FAM19A5 antagonist is an anti-FAM19A5 antibody or an antigen-binding fragment thereof, or a polynucleotide encoding an anti-FAM19A5 antibody, wherein the anti-FAM19A5 antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3, wherein (a) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 214, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 217, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (b) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 15, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 27, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 28; (c) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:207, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:208, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:213, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:28; (d) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 214, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 215, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (e) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 218, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (f) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:217, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:216; (g) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:219, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:220; or (h) the heavy chain CDR1 consists of the amino acid sequence shown in SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence shown in SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence shown in SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence shown in SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence shown in SEQ ID NO: 221, and the light chain CDR3 consists of the amino acid sequence shown in SEQ ID NO:

222.

8. The use according to claim 7, wherein the external stimulus is a mechanical stimulus, or a thermal stimulus, or both.

9. Use of a FAM19A5 antagonist for preparing a medicament for increasing sensory nerve conduction velocity in a subject in need thereof, wherein the FAM19A5 antagonist is an anti-FAM19A5 antibody or an antigen-binding fragment thereof, or a polynucleotide encoding an anti-FAM19A5 antibody, wherein the anti-FAM19A5 antibody comprises a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, wherein (a) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 214, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 217, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (b) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 15, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 27, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 28; (c) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:207, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:208, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:213, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:28; (d) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 214, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 215, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (e) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 218, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 216; (f) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:217, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:216; (g) the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO:209, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO:210, and the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO:16; the light chain CDR1 consists of the amino acid sequence of SEQ ID NO:26, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO:219, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:220; or (h) the heavy chain CDR1 consists of the amino acid sequence shown in SEQ ID NO: 209, the heavy chain CDR2 consists of the amino acid sequence shown in SEQ ID NO: 210, and the heavy chain CDR3 consists of the amino acid sequence shown in SEQ ID NO: 16; the light chain CDR1 consists of the amino acid sequence shown in SEQ ID NO: 26, the light chain CDR2 consists of the amino acid sequence shown in SEQ ID NO: 221, and the light chain CDR3 consists of the amino acid sequence shown in SEQ ID NO:

222.

10. The use according to any one of claims 1 to 9, wherein the anti-FAM19A5 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (1) VH comprises the amino acid sequence set forth in SEQ ID NO: 224 and VL comprises the amino acid sequence set forth in SEQ ID NO: 229; (2) VH comprises the amino acid sequence shown in SEQ ID NO:36 and VL comprises the amino acid sequence shown in SEQ ID NO:40; (3) VH comprises the amino acid sequence shown in SEQ ID NO: 223 and VL comprises the amino acid sequence shown in SEQ ID NO: SEQ ID NO: 227; (4) VH comprises the amino acid sequence shown in SEQ ID NO: 224 and VL comprises the amino acid sequence shown in SEQ ID NO: SEQ ID NO: 228; (5) VH comprises the amino acid sequence shown in SEQ ID NO: 224 and VL comprises the amino acid sequence shown in SEQ ID NO: SEQ ID NO: 230; (6) VH comprises the amino acid sequence shown in SEQ ID NO: 224 and VL comprises the amino acid sequence shown in SEQ ID NO: SEQ ID NO: 231; (7) VH comprises the amino acid sequence shown in SEQ ID NO: 224 and VL comprises the amino acid sequence shown in SEQ ID NO: SEQ ID NO: 232; or (8) VH comprises the amino acid sequence shown in SEQ ID NO:224 and VL comprises the amino acid sequence shown in SEQ ID NO:

233.

11. The use according to any one of claims 1 to 9, wherein the antigen binding fragment comprises Fab, Fab', F(ab')2, Fv single chain Fv (scFv) or a combination thereof.

12. The use according to any one of claims 1 to 9, wherein the anti-FAM19A5 antibody is a chimeric antibody, a human antibody or a humanized antibody.

13. The use according to any one of claims 1 to 9, wherein the FAM19A5 antagonist is linked to an agent to form an immunoconjugate.

14. The use according to any one of claims 1 to 9, wherein the FAM19A5 antagonist is formulated together with a pharmaceutically acceptable carrier.

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