Uses of c5 antibodies which bind to c5

Description

Uses of C5 antibodies that bind to C5 Separated from BR112018075688-1, filed on June 16, 2017. Technical Field

[001] The present invention relates to anti-C5 antibodies and methods of using them. Antecedent Technique

[002] The complement system plays a central role in the clearance of immune complexes and in immune responses to infectious agents, foreign antigens, virus-infected cells, and tumor cells. There are approximately 25 to 30 complement proteins, which are found as a complex collection of plasma proteins and membrane cofactors. Complement components achieve their immune defensive functions through interaction in a series of intricate enzymatic cleavages and membrane-binding events. The resulting complement cascades lead to the production of products with opsonistic, immunoregulatory, and lytic functions.

[003] Currently, it is widely accepted that the complement system can be activated through three distinct pathways: the classical pathway, the lectin pathway, and the alternative pathway. These pathways share several components and, despite differing in their initial steps, they converge and share the same terminal complement components (C5 to C9) responsible for the activation and destruction of target cells.

[004] The classical pathway is typically activated by the formation of antigen-antibody complexes. Regardless, the first step in lectin pathway activation is the binding of specific lectins, such as mannan-binding lectin (MBL), H-ficolin, M-ficolin, L-ficolin, and C-type lectin CL-11. In contrast, the alternative pathway spontaneously undergoes a low level of activation transfer, which can be readily amplified on foreign surfaces or Petition 870260079322, dated 07 / 08 / 2026, page 10 / 523 2 / 167 through abnormal surfaces (bacteria, yeast, virus-infected cells, or damaged tissue). These pathways converge at a point where the C3 component of complement is cleaved by an active protease to provide C3a and C3b.

[005] C3a is an anaphylatoxin. C3b binds to bacterial and other cells, as well as certain viruses and immune complexes, and marks them for removal from circulation (a role known as an opsonin). C3b also forms a complex with other components to form C5 convertase, which cleaves C5 into C5a and C5b.

[006] C5 is a 190 kDa protein found in normal serum at approximately 80 micrograms / ml (0.4 µM). C5 is glycosylated with about 1.5 to 3% of its mass attributed to carbohydrates. Mature C5 is a heterodimer with an alpha chain of 115 kDa that is disulfide-linked to a beta chain of 75 kDa. C5 is synthesized as a single-chain precursor protein (proC5 precursor) of 1676 amino acids (see, for example, PTL1 and PTL2). The pro-C5 precursor is cleaved to provide the beta chain as an amino-terminal fragment and the alpha chain as a carboxy-terminal fragment. The polypeptide fragments of the alpha and beta chains are linked to each other through a disulfide bond and constitute the mature C5 protein.

[007] Mature C5 is cleaved into the C5a and C5b fragments during activation of the complement pathways. C5a is cleaved from the alpha chain of C5 by C5 convertase as an amino-terminal fragment comprising the first 74 amino acids of the alpha chain. The remaining portion of mature C5 is the C5b fragment, which contains the remainder of the alpha chain disulfide-linked to the beta chain. Approximately 20% of the 11 kDa mass of C5a is attributed to carbohydrates.

[008] C5a is another anaphylatoxin. C5b combines with C6, C7, C8 and C9 to form the membrane attack complex (MAC, C5b Petition 870260079322, dated 07 / 08 / 2026, page 11 / 523 3 / 167 9, terminal complement complex (TCC)) on the surface of the target cell. When sufficient numbers of MACs are inserted into the membranes of target cells, MAC pores are formed to mediate rapid osmotic lysis of the target cells.

[009] As mentioned above, C3a and C5a are anaphylatoxins. They can trigger mast cell degranulation, which releases histamine and other inflammatory mediators, resulting in smooth muscle contraction, increased vascular permeability, leukocyte activation, and other inflammatory phenomena, including cell proliferation, resulting in hypercellularity. C5a also functions as a chemotactic peptide that serves to attract granulocytes, such as neutrophils, eosinophils, basophils, and monocytes, to the site of complement activation.

[0010] C5a activity is regulated by the plasma enzyme carboxypeptidase N, which removes the carboxy-terminal arginine from C5a, forming a C5a-des-Arg derivative. C5a-des-Arg exhibits only 1% of the anaphylactic activity and polymorphonuclear chemotactic activity of unmodified C5a.

[0011] Although a properly functioning complement system provides robust defense against infectious microbes, inadequate complement regulation or activation has been implicated in the pathogenesis of a variety of disorders, including, for example, rheumatoid arthritis (RA); lupus nephritis; ischemia-reperfusion injury; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration (e.g., age-related macular degeneration (AMD)); hemolytic syndrome due to elevated liver enzymes and low platelet count (HELLP); thrombotic thrombocytopenic purpura (TTP); spontaneous fetal loss; vasculitis Petition 870260079322, dated 07 / 08 / 2026, page 12 / 523 4 / 167 Pauci immune syndrome; epidermolysis bullosa; recurrent fetal loss; multiple sclerosis (MS); traumatic brain injury; and injury resulting from myocardial infarction, cardiopulmonary bypass, and hemodialysis (see, for example, NPL1). Therefore, inhibiting excessive or uncontrolled activations of the complement cascade may confer clinical benefits to patients with such disorders.

[0012] Paroxysmal nocturnal hemoglobinuria (PNH) is a rare hematological disorder in which red blood cells are compromised and therefore destroyed more rapidly than normal red blood cells. PNH results from the clonal expansion of hematopoietic stem cells with somatic mutations in the PIG-A gene (phosphatidylinositol glycan class A), which is located on the X chromosome. Mutations in PIG-A lead to an early blockage in the synthesis of glycosylphosphatidylinositol (GPI), a molecule that is necessary for the anchoring of various proteins to cell surfaces. Consequently, blood cells in PNH are deficient in GPI-anchored proteins, which include the complement regulatory proteins CD55 and CD59. Under normal circumstances, these complement regulatory proteins block the formation of MACs on cell surfaces, thus preventing erythrocyte lysis.The absence of GPI-anchored proteins causes complement-mediated hemolysis in PNH.

[0013] PNH is characterized by hemolytic anemia (a decreased number of red blood cells), hemoglobinuria (presence of hemoglobin in the urine, particularly evident after sleeping), and hemoglobinemia (presence of hemoglobin in the bloodstream). Individuals affected by PNH are known to experience paroxysms, which are defined here as the occurrence of dark-colored urine. Hemolytic anemia is due to the intravascular destruction of red blood cells by components of the virus. Petition 870260079322, dated 07 / 08 / 2026, page 13 / 523 5 / 167 supplement. Other known symptoms include dysphasia, fatigue, erectile dysfunction, thrombosis, and recurrent abdominal pain.

[0014] Eculizumab is a humanized monoclonal antibody directed against the C5 complement protein and the first approved therapy for the treatment of paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS) (see, for example, NPL2). Eculizumab inhibits the cleavage of C5 into C5a and C5b by C5 convertase, which prevents the generation of the terminal complement complex C5b-9. Both C5a and C5b cause the terminal complement-mediated events that are characteristic of PNH and aHUS (see also PTL3, PTL4, PTL5 and PTL6).

[0015] Several reports describe anti-C5 antibodies. For example, PTL7 described an anti-C5 antibody that binds to the alpha chain of C5 but does not bind to C5a, and blocks C5 activation, while PTL8 described a monoclonal anti-C5 antibody that inhibits C5a formation. On the other hand, PTL9 described an anti-C5 antibody that recognizes the proteolytic site for C5 convertase on the alpha chain of C5 and inhibits the conversion of C5 to C5a and C5b. PTL10 described an anti-C5 antibody that has a constant affinity of at least 1 x107M-1.

[0016] Antibodies (IgGs) bind to the neonatal Fc receptor (FcRn) and have long plasma retention times. IgG binding to FcRn is typically observed under acidic conditions (e.g., pH 6.0) and rarely under neutral conditions (e.g., pH 7.4). Typically, IgGs are incorporated non-specifically into cells via endocytosis and return to cell surfaces by binding to endosomal FcRn under acidic conditions in endosomes. Then, IgGs dissociate from FcRn under neutral conditions in plasma. IgGs that failed to bind to FcRn are degraded in lysosomes. When the binding capacity Petition 870260079322, dated 07 / 08 / 2026, p. 14 / 523 6 / 167 If the binding of an IgG to FcRn under acidic conditions is eliminated by introducing mutations in its Fc region, the IgG is not recycled from endosomes into the plasma, leading to a marked reduction in plasma IgG retention. To improve the plasma retention of IgGs, a method that enhances their binding to FcRn under acidic conditions has been reported. When the binding of an IgG to FcRn under acidic conditions is improved by introducing amino acid substitutions in its Fc region, the IgG is more efficiently recycled from endosomes to the plasma and thus shows enhanced plasma retention.However, it has also been reported that an IgG with enhanced binding to FcRn under neutral conditions does not dissociate from FcRn under neutral conditions in plasma, even when it returns to the cell surface through its binding to FcRn under acidic conditions in endosomes, and consequently, its retention time remains unchanged or, rather, is worsened (see, for example, NPL3, NPL4; NPL5).

[0017] Recently, antibodies that bind to antigens in a pH-dependent manner have been reported (see, for example, PTL1 and PTL2). These antibodies bind strongly to antigens under neutral plasma conditions and dissociate from antigens under acidic endosomal conditions. After dissociating from antigens, the antibodies become capable of binding to antigens again when recycled to the plasma via FcRn. Thus, a single antibody molecule can repeatedly bind to multiple antigen molecules. In general, the plasma retention of an antigen is much shorter than that of an antibody with the FcRn-mediated recycling mechanism mentioned above. Therefore, when an antigen is bound to an antibody, the antigen typically shows prolonged plasma retention, resulting in an increased plasma concentration of the antigen. On the other hand, Petition 870260079322, dated 07 / 08 / 2026, page 15 / 523 7 / 167 reported that the antibodies described above, which bind to antigens in a pH-dependent manner, clear antigens from plasma more rapidly than typical antibodies, since they dissociate from antigens within endosomes during the FcRn-mediated recycling process. PTL13 also described a computer modeling analysis showing that a pH-dependent binding antibody directed against C5 could prolong antigen knockdown. Citation List Patent Literature

[0018] [PTL1] United States Patent No6,355,245

[0019] [PTL2] United States Patent No7,432,356

[0020] [PTL3] WO2005 / 074607 [0021 ] [PTL4] WO2007 / 106585

[0022] [PTL5] WO2008 / 069889

[0023] [PTL6] WO2010 / 054403

[0024] [PTL7] WO 95 / 29697

[0025] [PTL8] WO 02 / 30985

[0026] [PTL9] WO2004 / 007553

[0027] [PTL10] WO2010 / 015608

[0028] [PTL11] WO2009 / 125825

[0029] [PTL12] WO2011 / 122011

[0030] [PTL13] WO2011 / 111007 Non-Patent Literature

[0031] [NPL1] Holers et al., Immunol. Rev. 223: 300-316 (2008)

[0032] [NPL2] Dmytrijuk et al., The Oncologist 13(9): 993-1000 (2008)

[0033] [NPL3] Yeung et al., J Immunol. 182(12): 7663-7671 (2009)

[0034] [NPL4] Datta-Mannan et al., J Biol. Chem. 282(3): 17091717 (2007) Petition 870260079322, dated 07 / 08 / 2026, p. 16 / 523 8 / 167

[0035] [NPL5] Dall'Acqua et al., J. Immunol. 169(9): 5171-5180 (2002) Summary of the Invention Technical Problem

[0036] One object of the invention is to provide anti-C5 antibodies and methods of using them. Solution to the Problem

[0037] The invention provides anti-C5 antibodies and methods for using them.

[0038] In some embodiments, an anti-C5 antibody isolated from the present invention binds to an epitope within the beta chain of C5. In some embodiments, an anti-C5 antibody isolated from the present invention binds to an epitope within the MG1-MG2 domain of the beta chain of C5. In some embodiments, an anti-C5 antibody isolated from the present invention binds to an epitope within a fragment consisting of amino acids 33-124 of the beta chain (SEQ ID NO: 40) of C5. In some embodiments, an anti-C5 antibody isolated from the present invention binds to an epitope within the beta chain (SEQ ID NO: 40) of C5 comprising at least one fragment selected from the group consisting of amino acids 47-57, 70-76 and 107-110.In some embodiments, an anti-C5 antibody isolated from the present invention binds to an epitope within the beta chain (SEQ ID NO: 40) of C5 comprising at least one amino acid residue selected from the group consisting of Glu48, Asp51, His70, His72, Lys109, and His110 of SEQ ID NO: 40. In further embodiments, the antibody binds to C5 with a higher affinity at a neutral pH than at an acidic pH. In further embodiments, the antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8. In another embodiment, an anti-C5 antibody isolated from the present invention binds to the same epitope as an antibody. Petition 870260079322, dated 07 / 08 / 2026, p. 17 / 523 9 / 167 described in Table 2. In further embodiments, the antibody binds to the same epitope as an antibody described in Table 2 with a higher affinity at pH 7.4 than at pH 5.8. In a further embodiment, an anti-C5 antibody isolated from the present invention binds to the same epitope as an antibody described in Tables 7 or 8. In further embodiments, the antibody binds to the same epitope as an antibody described in Tables 7 or 8 with a higher affinity at pH 7.4 than at pH 5.8.

[0039] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID NO: 1 and a VL of SEQ ID NO: 11; (b) a VH of SEQ ID NO: 5 and a VL of SEQ ID NO: 15; (c) a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 14; (d) a VH of SEQ ID NO: 6 and a VL of SEQ ID NO: 16; (e) a VH of SEQ ID NO: 2 and a VL of SEQ ID NO: 12; (f) a VH of SEQ ID NO: 3 and a VL of SEQ ID NO: 13; (g) a VH of SEQ ID NO: 9 and a VL of SEQ ID NO: 19; (h) a VH of SEQ ID NO: 7 and a VL of SEQ ID NO: 17; (i) a VH of SEQ ID NO: 8 and a VL of SEQ ID NO: 18; and (j) a VH of SEQ ID NO: 10 and a VL of SEQ ID NO: 10. In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a neutral pH than at an acidic pH. In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[0040] In some embodiments, an anti-C5 antibody of the present invention has a selected group feature consisting of: (a) the antibody contacts amino acids D51 and K109 of C5 (SEQ ID NO: 39); (b) the antibody's affinity for C5 (SEQ ID NO: 39) is greater than the antibody's affinity for a C5 mutant consisting of an E48A substitution of SEQ ID NO: 39; or (c) the anti Petition 870260079322, dated 07 / 08 / 2026, p. 18 / 523 10 / 167 The body binds to a C5 protein consisting of the amino acid sequence SEQ ID NO: 39 at a pH of 7.4, but does not bind to a C5 protein consisting of the amino acid sequence SEQ ID NO: 39 with an H72Y substitution at a pH of 7.4. In further embodiments, the antibody binds to C5 with a higher affinity at a neutral pH than at an acidic pH. In further embodiments, the antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[0041] In some embodiments, an anti-C5 antibody isolated from the present invention inhibits C5 activation. In some embodiments, an anti-C5 antibody isolated from the present invention inhibits the activation of the R885H variant of C5. In some embodiments, an anti-C5 antibody isolated from the present invention is a monoclonal antibody. In some embodiments, an anti-C5 antibody isolated from the present invention is a human, humanized, or chimeric antibody. In some embodiments, an anti-C5 antibody isolated from the present invention is a C5-binding antibody fragment. In some embodiments, an anti-C5 antibody isolated from the present invention is a full-length IgG1 or IgG4 antibody.

[0042] In some embodiments, an anti-C5 antibody isolated from the present invention comprises (a) an HVR-H3 comprising the amino acid sequence DX1GYX2X3PTHAMX4X5, wherein X1 is G or A, X2 is V, Q or D, X3 is T or Y, X4 is Y or H, X5 is L or Y (SEQ ID NO: 128), (b) an HVR-L3 comprising the amino acid sequence QX1TX2VGSSYGNX3, wherein X1 is S, C, N or T, X2 is F or K, X3 is A, T or H (SEQ ID NO: 131), and (c) an HVR-H2 comprising the amino acid sequence X1IX2TGSGAX3YX4AX5WX6KG, wherein X1 is C, A or G, X2 is Y or F, X3 is T, D or E, X4 is Y, K or Q, X5 is S, D or And, X6 is A or V (SEQ ID NO: 127).

[0043] In some embodiments, an anti-C5 antibody isolated from Petition 870260079322, dated 07 / 08 / 2026, p. 19 / 523 11 / 167 The present invention comprises (a) an HVR-H1 comprising the amino acid sequence SSYYX1X2, wherein X1 is M or V, X2 is C or A (SEQ ID NO: 126), (b) an HVR-H2 comprising the amino acid sequence X1IX2TGSGAX3YX4AX5WX6KG, wherein X1 is C, A or G, X2 is Y or F, X3 is T, D or E, X4 is Y, K or Q, X5 is S, D or E, X6 is A or V (SEQ ID NO: 127) and (c) an HVR-H3 comprising the amino acid sequence DX1GYX2X3PTHAMX4X5, wherein X1 is G or A, X2 is V, Q or D, X3 is T or Y, X4 is Y or H, X5 is L or Y (SEQ ID NO: 128).In further embodiments, the antibody comprises (a) an HVR-L1 comprising the amino acid sequence X1ASQX2IX3SX4LA, wherein X1 is Q or R, X2 is N, Q or G, X3 is G or S, X4 is D, K or S (SEQ ID NO: 129); (b) an HVR-L2 comprising the amino acid sequence GASX1X2X3S, wherein X1 is K, E or T, X2 is L or T, X3 is A, H, E or Q (SEQ ID NO: 130); and (c) an HVR-L3 comprising the amino acid sequence QX1TX2VGSSYGNX3, wherein X1 is S, C, N or T, X2 is F or K, X3 is A, T or H (SEQ ID NO: 131).

[0044] In some embodiments, an anti-C5 antibody isolated from the present invention comprises (a) an HVR-L1 comprising the amino acid sequence X1ASQX2IX3SX4LA, where X1 is Q or R, X2 is N, Q or G, X3 is G or S, X4 is D, K or S (SEQ ID NO: 129); (b) an HVR-L2 comprising the amino acid sequence GASX1X2X3S, where X1 is K, E or T, X2 is L or T, X3 is A, H, E or Q (SEQ ID NO: 130); and (c) an HVR-L3 comprising the amino acid sequence QX1TX2VGSSYGNX3, where X1 is S, C, N or T, X2 is F or K, X3 is A, T or H (SEQ ID NO: 131).

[0045] In some embodiments, an anti-C5 antibody isolated from the present invention comprises a structural region FR1 of the variable heavy chain domain comprising the amino acid sequence of any of the SEQ ID NOs: 132-134; FR2 comprising the amino acid sequence of any of the SEQ ID NOs: 135 Petition 870260079322, dated 07 / 08 / 2026, p. 20 / 523 12 / 167 136; FR3 comprising the amino acid sequence of any of the SEQ ID NOs: 137-139; and FR4 comprising the amino acid sequence of any of the SEQ ID NOs: 140-141. In some embodiments, an anti-C5 antibody isolated from the present invention comprises a structural region FR1 of the variable light chain domain comprising the amino acid sequence of any of the SEQ ID NOs: 142-143; FR2 comprising the amino acid sequence of any of the SEQ ID NOs: 144-145; FR3 comprising the amino acid sequence of any of the SEQ ID NOs: 146-147; and FR4 comprising the amino acid sequence of SEQ ID NO: 148.

[0046] In some embodiments, an anti-C5 antibody isolated from the present invention comprises (a) a VH sequence having at least 95% sequence identity with the amino acid sequence of any of SEQ ID NOs: 10, 106-110; (b) a VL sequence having at least 95% sequence identity with the amino acid sequence of any of SEQ ID NOs: 20, 111-113; or (c) a VH sequence as in (a) and a VL sequence as in (b). In further embodiments, the antibody comprises a VH sequence of any of SEQ ID NOs: 10, 106-110. In further embodiments, the antibody comprises a VL sequence of any of SEQ ID NOs: 20, 111-113.

[0047] The invention provides an antibody comprising a VH sequence from any of SEQ ID NOs: 10, 106-110 and a VL sequence from any of SEQ ID NOs: 20, 111-113.

[0048] The invention also provides isolated nucleic acids encoding an anti-C5 antibody of the present invention. The invention also provides host cells comprising a nucleic acid of the present invention. The invention also provides a method Petition 870260079322, dated 07 / 08 / 2026, page 21 / 523 13 / 167 of producing an antibody comprising cultivating a host cell of the present invention so that the antibody is produced.

[0049] The invention further provides a method for producing an anti-C5 antibody. In some embodiments, the method comprises immunizing an animal against a polypeptide comprising the MG1-MG2 domain (SEQ ID NO: 43) of the beta chain of C5. In some embodiments, the method comprises immunizing an animal against a polypeptide comprising the region corresponding to amino acids at positions 33 to 124 of the beta chain (SEQ ID NO: 40) of C5. In some embodiments, the method comprises immunizing an animal against a polypeptide comprising at least one fragment selected from amino acids 47-57, 70-76 and 107-110 of the beta chain (SEQ ID NO: 40) of C5. In some embodiments, the method involves immunizing an animal against a polypeptide comprising a beta-chain fragment (SEQ ID NO: 40) of C5 comprising at least one amino acid selected from Glu48, Asp51, His70, His72, Lys109, and His110.

[0050] The invention also provides a pharmaceutical formulation comprising an anti-C5 antibody of the present invention and a pharmaceutically acceptable carrier.

[0051] The anti-C5 antibodies of the present invention can be used as a medicine. The anti-C5 antibodies of the present invention can be used in the treatment of a complement-mediated disease or condition involving excessive or uncontrolled activation of C5. The anti-C5 antibodies of the present invention can be used to increase the clearance of C5 from plasma.

[0052] The anti-C5 antibodies of the present invention can be used in the manufacture of a medicament. In some embodiments, the medicament is for the treatment of a disease or condition me Petition 870260079322, dated 07 / 08 / 2026, page 22 / 523 14 / 167 diated by complement involving excessive or uncontrolled activation of C5. In some modalities, the drug is to improve the elimination of C5 from the plasma.

[0053] The invention also provides a method for treating an individual with a complement-mediated disease or condition involving excessive or uncontrolled C5 activation. In some embodiments, the method comprises administering to the individual an effective amount of an anti-C5 antibody of the present invention. The invention also provides a method for increasing the clearance of C5 from plasma in an individual. In some embodiments, the method comprises administering to the individual an effective amount of an anti-C5 antibody of the present invention to increase the clearance of C5 from plasma. Brief Description of the Drawings

[0054] Figure 1 illustrates the binning of anti-C5 antibody epitopes, as described in Example 2.2. Antibodies grouped at the same epitope site are framed with a thick line.

[0055] Figure 2A illustrates BIACORE (registered trademark) sensorgrams of anti-C5 antibodies at a pH of 7.4 (solid line) and pH of 5.8 (dashed line) to assess pH dependence, as described in Example 3.2. CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, and CFA0599 are antibodies clustered at the C epitope, as described in Example 2.2.

[0056] Figure 2B illustrates BIACORE (registered trademark) sensorgrams of anti-C5 antibodies at a pH of 7.4 (solid line) and pH of 5.8 (dashed line) to assess pH dependence, as described in Example 3.2. CFA0666, CFA0672, and CFA0675 are antibodies clustered at the C epitope, and CFA0330 and CFA0341 are antibodies clustered at the B epitope, as described in Example 2.2. 305LO5 is a Petition 870260079322, dated 07 / 08 / 2026, page 23 / 523 15 / 167 humanized antibody of CFA0305, as described in Example 2.3.

[0057] Figure 3 illustrates Western blot analysis against C5 beta-chain derived fragments (amino acids 19-180, 161-340, 321-500, and 481-660 of SEQ ID NO: 40) fused to a GST marker, as described in Example 4.1. CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675 are antibodies clustered at the C epitope. The anti-GST antibody is a positive control. The position of the GST-fused C5 fragments (46-49 kDa) is marked with an arrow.

[0058] Figure 4 illustrates BIACORE (registered trademark) sensorgrams of anti-C5 antibodies against the MG1-MG2 domain of the C5 beta chain, as described in Example 4.3. The upper panel shows the results for CFA0305 (solid line), CFA0307 (dashed line), CFA0366 (dash-dotted line) and eculizumab (dotted line), CFA0538 (dash-dotted line) and eculizumab (dotted line). The lower panel shows the results for CFA0666 (solid line), CFA0672 (dashed line), CFA0675 (dash-dotted line) and eculizumab (dotted line). CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675 are antibodies grouped at the C epitope. Eculizumab is a control anti-C5 antibody.

[0059] Figure 5A illustrates Western Blot analysis against peptide fragments derived from the MG1-MG2 domain (amino acids 33-124, 45-124, 52-124, 33-111, 33-108 and 45-111 of SEQ ID NO: 40) fused with a GST marker, as described in Example 4.4. The anti-GST antibody is used as an antibody for the reaction. The position of the C5 fragments fused to GST (35-37 kDa) is marked with an arrow.

[0060] Figure 5B illustrates the Western Blot analysis against fragments Petition 870260079322, dated 07 / 08 / 2026, page 24 / 523 16 / 167 peptide groups derived from the MG1-MG2 domain (amino acids 33-124, 45-124, 52-124, 33-111, 33-108 and 45-111 of SEQ ID NO: 40) fused with a GST marker, as described in Example 4.4. CFA0305 is used as an antibody for the reaction.

[0061] Figure 5C summarizes the binding reactions of anti-C5 antibodies to fragments derived from the beta chain of C5, as described in Example 4.4. The fragments to which the C-epitope-grouped anti-C5 antibodies (CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675) bind are shown in gray, and the fragments to which they do not bind are shown in white.

[0062] Figure 6 illustrates the Western Blot analysis against C5 point mutants, in which E48, D51, and K109 of the beta chain are substituted for alanine (E48A, D51A, and K109A, respectively), as described in Example 4.5. In the left panel, eculizumab (anti-C5 antibody, alpha chain ligand) is used as an antibody for the reaction, and the position of the C5 alpha chain (approximately 113 kDa) is marked with an arrow. In the right panel, CFA0305 (clustered epitope C, beta chain ligand) is used as an antibody for the reaction, and the position of the C5 beta chain (approximately 74 kDa) is marked with an arrow.

[0063] Figure 7 illustrates BIACORE (registered trademark) sensorgrams showing the interaction of eculizumab-F760G4 (top panel) or 305LO5 (bottom panel) with C5 mutants, as described in Example 4.6. The sensorgrams were obtained by injecting C5-wt (thick solid curve), C5-E48A (short dashed curve), C5-D51A (long dashed curve), and C5-K109A (thin solid curve), respectively, onto the surface of the sensor immobilized with eculizumab-F760G4 or 305LO5. Eculizumab is a control anti-C5 antibody. 305LO5 is a humanized antibody of CFA0305 (clustered at the C epitope). Petition 870260079322, dated 07 / 08 / 2026, page 25 / 523 17 / 167 as described in Example 2.3.

[0064] Figure 8 illustrates BIACORE (registered trademark) sensorgrams showing the interaction of 305LO5 with C5 His mutants to assess pH dependence, as described in Example 4.7. The sensorgrams were obtained by injecting C5-wt (thick solid curve), C5-H70Y (long dashed curve), C5-H72Y (short dashed curve), C5-H110Y (dotted curve), and C5-H70Y+H110Y (thin solid curve), respectively, onto the surface of the sensor immobilized with 305LO5. The antibody / antigen complexes were allowed to dissociate at pH 7.4, followed by further dissociation at pH 5.8 (indicated with an arrow) to assess pH-dependent interactions.

[0065] Figure 9A illustrates the inhibition of complement-activated liposome lysis by anti-C5 antibodies, as described in Example 5.1. Results are shown for CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675 grouped at epitope C, as described in Example 2.2.

[0066] Figure 9B illustrates the inhibition of complement-activated liposome lysis by anti-C5 antibodies, as described in Example 5.1. Results are shown for the CFA0330 and CFA0341 antibodies clustered at epitope B, as described in Example 2.2.

[0067] Figure 10A illustrates the inhibition of C5 generation by anti-C5 antibodies, as described in Example 5.2. C5a concentrations were quantified in the supernatants obtained during the liposome lysis assay described in Figure 9A.

[0068] Figure 10B illustrates the inhibition of C5 generation by anti-C5 antibodies, as described in Example 5.2. C5a concentrations were quantified in the supernatants obtained during the liposome lysis assay described in Figure 9B. Petition 870260079322, dated 07 / 08 / 2026, page 26 / 523 18 / 167

[0069] Figure 11 illustrates the inhibition of complement-activated hemolysis by anti-C5 antibodies, as described in Example 5.3. The complements were activated via the classical pathway.

[0070] Figure 12 illustrates the inhibition of complement-activated hemolysis by anti-C5 antibodies, as described in Example 5.4. The complements were activated via the alternative pathway.

[0071] Figure 13 illustrates the time course of C5 concentration in human plasma after intravenous administration of human C5 alone or human C5 and an anti-human C5 antibody in mice to assess C5 clearance, as described in Example 6.2. CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675 antibodies clustered at epitope C and CFA0330 and CFA0341 antibodies clustered at epitope B, as described in Example 2.2.

[0072] Figure 14 illustrates the time course of the concentration of human anti-C5 antibody in plasma after intravenous administration of human C5 and a human anti-C5 antibody in mice to evaluate antibody pharmacokinetics, as described in Example 6.3. CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675 antibodies clustered at epitope C and CFA0330 and CFA0341 antibodies clustered at epitope B, as described in Example 2.2.

[0073] Figure 15 illustrates the inhibition of complement-activated liposome lysis by anti-C5 antibodies, as described in Example 9.1. Results are shown for antibodies 305LO15-SG422, 305LO16-SG422, 305LO18-SG422, 305LO19SG422, 305LO20-SG422, and 305LO20-SG115.

[0074] Figure 16 illustrates the inhibition of complement-activated liposome lysis by anti-C5 antibodies, as described in Example 9.1. The results for the antibodies are shown. Petition 870260079322, dated 07 / 08 / 2026, page 27 / 523 19 / 167 305LO15-SG115 and 305LO23-SG429.

[0075] Figure 17 illustrates the inhibition of complement-activated liposome lysis by anti-C5 antibodies, as described in Example 9.1. Results are shown for antibodies 305LO22-SG115, 305LO22-SG422, 305LO23-SG115, and 305LO23SG422.

[0076] Figure 18 illustrates the inhibition of C5a generation by anti-C5 antibodies, as described in Example 9.2. C5a concentrations were quantified in the supernatants obtained during the liposome lysis assay described in Figure 15.

[0077] Figure 19 illustrates the inhibition of C5a generation by anti-C5 antibodies, as described in Example 9.2. C5a concentrations were quantified in the supernatants obtained during the liposome lysis assay described in Figure 16.

[0078] Figure 20 illustrates the inhibition of complement activity in monkey plasma by anti-C5 antibodies, as described in Example 9.3. Anti-C5 antibodies were administered to cynomolgus monkeys and complement activities in monkey plasma were measured in a hemolysis assay.

[0079] Figure 21 illustrates the inhibition of the biological activity of wild-type (WT) C5 and C5 variants (V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N, and E1437D) by an anti-C5 antibody (eculizumab), as described in Example 9.4.

[0080] Figure 22 illustrates the inhibition of the biological activity of wild-type (WT) C5 and C5 variants (V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N, and E1437D) by an anti-C5 antibody (a 305 variant), as described in Example 9.4.

[0081] Figure 23 illustrates the inhibition of complement-activated liposome lysis by anti-C5 antibodies (BNJ441 and a variant of 305), as described in Example 9.5. Petition 870260079322, dated 07 / 08 / 2026, page 28 / 523 20 / 167

[0082] Figure 24 illustrates the time course of cynomolgus C5 concentration in plasma after intravenous administration of an anti-human C5 antibody in cynomolgus monkeys to assess C5 clearance, as described in Example 10.2.

[0083] Figure 25 illustrates the time course of human anti-C5 antibody concentration in plasma after intravenous administration of a human anti-C5 antibody in cynomolgus monkeys to evaluate antibody pharmacokinetics, as described in Example 10.3.

[0084] Figures 26A and 26B illustrate the crystal structure of Fab 305 bound to the MG1 domain of human C5 (hC5), as described in Example 11.6. Figure 26A illustrates an asymmetric unit. MG1 is shown in a surface representation and Fab 305 is shown as ribbons (dark gray: heavy chain, light gray: light chain). Figure 26B illustrates overlapping molecules 1 and 2 (dark gray: molecule 1, light gray: molecule 2).

[0085] Figure 27A illustrates the Fab 305 contact region epitope on the MG1 domain, as described in Example 11.6. Figure 27A illustrates the mapping of the epitope to the MG1 amino acid sequence (dark gray: closer than 3.0 Angstrom, light gray: closer than 4.5 Angstrom).

[0086] Figure 27B illustrates the contact region epitope of Fab 305 over the MG1 domain, as described in Example 11.6. Figure 27B illustrates the mapping of the epitope onto the crystal structure (dark gray spheres: closer than 3.0 Angstrom, light gray rods: closer than 4.5 Angstrom).

[0087] Figure 28A illustrates a foreground visualization of the interactions of E48, D51, and K109 (rod representation) with Fab 305 (surface representation), as described in Example 11.7.

[0088] Figure 28B illustrates the interactions between E48 and its environment. Petition 870260079322, dated 07 / 08 / 2026, page 29 / 523 21 / 167 te (dark gray dashed line: hydrogen bonding with Fab, light gray dashed line: water-mediated hydrogen bonding), as described in Example 11.7.

[0089] Figure 28C illustrates the interactions between D51 and its environment (dark gray dashed line: hydrogen bonding with Fab), as described in Example 11.7.

[0090] Figure 28D illustrates the interactions between K109 and its environment (dark gray dashed line: hydrogen bonding with Fab, light gray dashed line: salt bridge with H-CDR3_D95), as described in Example 11.7.

[0091] Figure 29A illustrates an enlarged view of the interactions of H70, H72, and H110 (rod representation) with Fab 305 (surface representation), as described in Example 11.8, in the same orientation as Figure 28A.

[0092] Figure 29B illustrates the interactions between H70 and its environment, as described in Example 11.8. This histidine residue is indicated in a stem-and-lattice representation. The hydrogen bond is indicated by the dashed line.

[0093] Figure 29C illustrates the interactions between H72 and its environment, as described in Example 11.8. This histidine residue is indicated in a stem-and-lattice representation. The hydrogen bond is indicated by the dashed line.

[0094] Figure 29D illustrates the interactions between H110 and its environment, as described in Example 11.8. This histidine residue is shown in a stem-and-lattice representation. The hydrogen bond is indicated by the dashed line. The distance between H110 and HCDR3_H100c is shown by the dashed line. Description of the Modalities

[0095] The techniques and procedures described or cited herein are generally well understood and commonly employed using Petition 870260079322, dated 07 / 08 / 2026, p. 30 / 523 22 / 167 uma metodologia convencional por aqueles versados na técnica tais como, por exemplo, as metodologias amplamente usadas descritas em Sambrook et al., Molecular Cloning: A Laboratory Manual, 3aedição (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology (F.M. Ausubel et al. eds., (2003)); a série Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames and G.R. Taylor eds. (1995)), Harlow and Lane, eds. (1988) Antibodies, A Laboratory Manual and Animal Cell Culture (R.I. Freshney, ed. (1987)); Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney), ed., 1987); Introduction to Cell and Tissue Culture (J. P. Mather e P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths e D.G.Newell, eds., 1993-8) J. Wiley and Sons; Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos, eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al. eds., 1994); Current Protocols in Immunology (J.E. Coligan et al. eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway e P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty., ed., IRL Press, 1988-1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean eds., Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow e D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J. D. Capra eds., Harwood Academic Publishers, 1995); e Cancer: Principles and Practice of Oncology (V.T. DeVita et al. eds., J.B. Lippincott Company, 1993). DEFINIÇÕES Petition 870260079322, dated 07 / 08 / 2026, p. 31 / 523 23 / 167

[0096] Unless defined otherwise, the technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. Singleton et al., Dictionary of Microbiology and Molecular Biology, 2nd ed., J. Wiley & Sons (New York, NY 1994) and March, Advanced Organic Chemistry Reactions, Mechanisms and Structure, 4th ed., John Wiley & Sons (New York, NY 1992) provide for those skilled in the art a general guide to several of the expressions used in the present application. All references cited herein, including patent applications and publications, are incorporated by reference in full.

[0097] For the purposes of interpreting this application, the following definitions shall apply and, where appropriate, expressions used in the singular shall also include the plural and vice versa. It should be understood that the terminology used herein is intended only to describe particular modalities and is not intended to be limiting. In the event that any definition described below conflicts with any document incorporated herein by reference, the definition described below shall prevail.

[0098] A human acceptor structural region, for the purposes hereof, is a structural region comprising the amino acid sequence of the structural region of a variable light chain (VL) domain or the structural region of a variable heavy chain (VH) domain derived from a human immunoglobulin structural region or a human consensus structural region, as defined below. A human acceptor structural region derived from a human immunoglobulin structural region or a human consensus structural region may comprise the same amino acid sequence as the same or may contain alterations in the amino acid sequence. In some embodiments, the Petition 870260079322, dated 07 / 08 / 2026, page 32 / 523 24 / 167 The number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the human VL acceptor structural region has a sequence identical to the human immunoglobulin VL structural region sequence or the human consensus structural region sequence.

[0099] Affinity refers to the intensity of the total 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, as used herein, binding affinity refers to intrinsic binding affinity which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can be represented, in general, by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[00100] A matured affinity antibody refers to an antibody with one or more alterations in one or more hypervariable regions (HVRs), compared to a parental antibody that does not have such alterations, such alterations resulting in an enhancement in the antibody's affinity for the antigen.

[00101] The terms anti-C5 antibody and an antibody that binds to C5 refer to an antibody that is capable of binding to C5 with sufficient affinity, such that the antibody is useful as a diagnostic and / or therapeutic agent in targeting C5. In one embodiment, the extent of binding of an anti-C5 antibody to an unrelated non-C5 protein is less than about 10% of the antibody binding to C5 as measured, for example, by a ra Petition 870260079322, dated 07 / 08 / 2026, p. 33 / 523 25 / 167 immunoassay (RIA). In certain embodiments, an antibody that binds to C5 has a dissociation constant (Kd) of 1 microM or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less, or 0.001 nM or less (e.g., 10⁻⁸M or less, e.g., from 10⁻⁸M to 10⁻¹³M, e.g., from 10⁻⁹M to 10⁻¹³M). In certain embodiments, an anti-C5 antibody binds to a C5 epitope that is conserved among C5 from different species.

[00102] The term antibody is used here in the broadest sense and encompasses various antibody structures including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, provided they exhibit the desired antigen-binding activity.

[00103] An antibody fragment refers to a molecule other than an intact antibody comprising a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., ScFv); and multispecific antibodies formed from antibody fragments.

[00104] An antibody that binds to the same epitope as a reference antibody refers to an antibody that blocks the binding of the reference antibody to its antigen in a competition assay and / or, conversely, the reference antibody blocks the binding of the antibody to its antigen in a competition assay. An example competition assay is provided here.

[00105] The term chimeric antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a source Petition 870260079322, dated 07 / 08 / 2026, p. 34 / 523 26 / 167 or a particular species, while the remainder of the light and / or heavy chain is derived from a different source or species.

[00106] The class of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The constant domains of the heavy chain that correspond to the different classes of immunoglobulins are designated alpha, delta, epsilon, gamma, and mu, respectively.

[00107] The term cytotoxic agent, as used herein, refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212 and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth-inhibiting agents; enzymes and fragments thereof, such as nucleolytic enzymes; antibiotics; Toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and the various antitumor and anticancer agents described below.

[00108] Effector functions refer to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: binding to C1q and antibody-dependent cytotoxicity (CDC); binding to the Fc receptor; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; negative regulation Petition 870260079322, dated 07 / 08 / 2026, p. 35 / 523 27 / 167 activation of receptors on the cell surface (e.g., B cell receptor); and activation of B cells.

[00109] An effective amount of an agent, for example, a pharmaceutical formulation, refers to an effective quantity, in dosages and during periods of time necessary, to obtain the desired therapeutic or prophylactic result.

[00110] The term epitope includes any determinant capable of being bound by an antibody. An epitope is a region of an antigen that is bound by an antibody targeting that antigen and includes specific amino acids that directly contact the antibody. Epitope determinants may include chemically active surface clusters of molecules, such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and may have specific three-dimensional structural features and / or specific charge characteristics. In general, antibodies specific for a particular target antigen will preferentially recognize an epitope on a target antigen in a complex mixture of proteins and / or macromolecules.

[00111] The term Fc region is used here to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The expression includes native sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or the constant region is in accordance with the EU numbering system, also referred to as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, Na Petition 870260079322, dated 07 / 08 / 2026, page 36 / 523 28 / 167 tional Institutes of Health, Bethesda, MD, 1991.

[00112] Structural region or FR refers to the residues of the variable domain that are different from the residues of the hypervariable region (HVR). The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Consequently, the HVR and FR sequences generally appear in the following sequence in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[00113] The terms full-length antibody, intact antibody, and complete antibody are used here interchangeably to refer to an antibody that has a structure substantially similar to a native antibody structure or that has heavy chains containing an Fc region as defined herein.

[00114] The terms host cell, host cell lineage, and host cell culture are used interchangeably and refer to cells into which an exogenous nucleic acid has been introduced, including the progeny of such cells. The host cell includes transformants and transformed cells, which include the primary transformed cell and the progeny derived from it regardless of the number of passages. The progeny may not be completely identical, as to nucleic acid content, to a parent cell, but may contain mutations. Mutant progeny that has the same biological function or activity as screened for or selected in the originally transformed cell is included here.

[00115] A human antibody is one that has an amino acid sequence that corresponds to that of an antibody produced by a human being or a human cell or derived from a non-human source that uses human antibody repertoires or other sequences that encode a human antibody. This definition of a human antibody specifically excludes a humanized antibody comprising residues that bind to a non-human antigen. Petition 870260079322, dated 07 / 08 / 2026, p. 37 / 523 29 / 167

[00116] A human consensus structural region is a structural region that represents the amino acid residues that occur most commonly in a selection of human immunoglobulin VL or VH structural sequences. In general, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. In general, the sequence subgroup is a subgroup as described in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), volumes 1-3. In one embodiment, for VL, the subgroup is the kappa I subgroup as described in Kabat et al., supra. In one embodiment, for VH, the subgroup is the III subgroup as described in Kabat et al., supra.

[00117] A humanized antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise all of at least one and, typically, two variable domains, in which all or substantially all HVRs (e.g., CDRs) correspond to those of a non-human antibody and all or substantially all FRs correspond to those of a human antibody. A humanized antibody may optionally comprise at least a portion of a constant region derived from a human antibody. The humanized form of an antibody, e.g., a non-human antibody, refers to an antibody that has undergone humanization.

[00118] The term hypervariable region or HVR, as used herein, refers to each of the regions of a variable domain of an antibody that are of hypervariable sequence (complementarity-determining regions or CDRs) and / or form structurally defined loops (hypervariable loops) and / or contain residues that contact the antigen (antigen contacts). In general, antibodies. Petition 870260079322, dated 07 / 08 / 2026, page 38 / 523 30 / 167 comprise six HVRs: three in VH (H1, H2, H3) and three in VL (L1, L2, L3). Exemplary HVRs include: (a) hypervariable loops that occur at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia, J. Mol. Biol. 196: 901-917 (1987)); (b) CDRs that occur at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, NIH, Bethesda, MD (1991)); (c) antigen contacts that occur at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al., J. Mol. Biol. 262: 732-745 (1996)); and (d) combinations of (a), (b) and / or (c), including amino acid residues from HVR 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3) and 94-102 (H3).

[00119] Unless otherwise indicated, HVR residuals and other residuals in the variable domain (e.g., FR residuals) are numbered here in accordance with Kabat et al., supra.

[00120] An immunoconjugate is an antibody conjugated to one or more heterologous molecules including, but not limited to, a cytotoxic agent.

[00121] An individual or patient is a mammal. Mammals include, but are not limited to, domestic animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain modalities, the individual or patient is a human being.

[00122] An isolated antibody is one that has been separated from a component of its natural environment. In some embodiments, an antibody is purified to more than 95% or 99% purity as determined, for example, by electrophoretic methods. Petition 870260079322, dated 07 / 08 / 2026, page 39 / 523 31 / 167 cos (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatographic (e.g., ion exchange or reverse-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:7 9-87 (2007).

[00123] An isolated nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that commonly contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or in a chromosomal location that is different from its natural chromosomal location.

[00124] Isolated nucleic acid encoding an anti-C5 antibody refers to one or more nucleic acid molecules encoding the heavy and light chains of the antibody (or fragments thereof), including such nucleic acid molecules in a single vector or separate vectors and such nucleic acid molecules present in one or more locations in a host cell.

[00125] The term monoclonal antibody, as used herein, refers to an antibody obtained from a substantially homogeneous antibody population, that is, the individual antibodies comprising the population are identical and / or bind to the same epitope, except for possible variant antibodies, for example, those containing mutations that occur naturally or that arise during the production of a monoclonal antibody preparation, such variants generally being present in small quantities. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single determinant on Petition 870260079322, dated 07 / 08 / 2026, page 40 / 523 32 / 167 an antigen. Therefore, the monoclonal modifier indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be considered as requiring the production of the antibody through any particular method. For example, the monoclonal antibodies to be used according to the present invention can be made by means of a variety of techniques including, but not limited to, hybridoma methods, recombinant DNA methods, phage display methods and methods using transgenic animals containing all or part of the human immunoglobulin loci, such methods and other exemplary methods for producing monoclonal antibodies being described herein.

[00126] A nude antibody refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or radioactive marker. The nude antibody may be present in a pharmaceutical formulation.

[00127] Native antibodies refer to naturally occurring immunoglobulin molecules with variable structures. For example, native IgG antibodies are heterodimeric glycoproteins of approximately 150,000 daltons composed of two identical light chains and two identical heavy chains that are disulfide-linked. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH), also called a heavy variable domain or a heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also called a light variable domain or light chain variable domain, followed by a light constant domain (CL). The light chain of an antibody can be designated in one of two types, called kappa (kappa) and lambda (lambda), based on the amino acid sequence of its... Petition 870260079322, dated 07 / 08 / 2026, page 41 / 523 33 / 167 constant minimum.

[00128] The expression "inserted in the packaging" is used to refer to instructions commonly included in commercial packaging of therapeutic products, which contain information about the indications, use, dosage, administration, combination therapy, contraindications and / or warnings regarding the use of such therapeutic products.

[00129] Percentage (%) of amino acid sequence identity, relative to a reference polypeptide sequence, is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence after sequence alignment and introduction of gaps, if necessary, to obtain the maximum percentage of sequence identity and not considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining the percentage of amino acid sequence identity can be achieved in several ways that are within the knowledge of the art, for example, using publicly available computer programs such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) programs.Those skilled in the art can determine the appropriate parameters for sequence alignment, including any algorithms necessary to achieve maximum alignment over the full length of the sequences to be compared. For the purposes herein, however, the % amino acid sequence identity values ​​are generated using the ALIGN-2 sequence comparison computer program. The ALIGN-2 sequence comparison computer program is licensed by Genentech, Inc., and the source code has been deposited with the user documentation at the US Copyright Office, Washington DC, 20559, where it is registered as [reference to copyright registration number]. Petition 870260079322, dated 07 / 08 / 2026, page 42 / 523 34 / 167 US Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or can be compiled from source code. The ALIGN-2 program must be compiled for use on a UNIX operating system, including UNIX V4.0D digital. All sequence comparison parameters are described in the ALIGN-2 program and do not vary.

[00130] In situations where ALIGN-2 is used for amino acid sequence comparisons, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which may alternatively be written as a given amino acid sequence A having or comprising a given % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 times the fraction X / Y, where X is the number of amino acid residues classified as identical combinations by the ALIGN-2 sequence alignment program characterized by the alignment program of A and B and where Y is the total number of amino acid residues in B.It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity from A to B will not be equal to the % amino acid sequence identity from B to A. Unless specifically determined otherwise, all % amino acid sequence identity values ​​used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[00131] The term pharmaceutical formulation refers to a preparation that is in such a form as to allow the biological activity of an active ingredient contained therein to be effective and that does not contain additional components that are unacceptably toxic. Petition 870260079322, dated 07 / 08 / 2026, p. 43 / 523 35 / 167 cos for an individual to whom the formulation will be administered.

[00132] A pharmaceutically acceptable vehicle refers to an ingredient in a pharmaceutical formulation, other than the active ingredient, which is not toxic to the individual. A pharmaceutically acceptable vehicle includes, but is not limited to, a buffer, excipient, stabilizer, or preservative.

[00133] The term C5, as used herein, encompasses any native C5 from any vertebrate source, including mammals such as primates (e.g., humans and monkeys) and rodents (e.g., mice and rats). Unless otherwise indicated, the term C5 refers to human C5 protein having the amino acid sequence shown in SEQ ID NO: 39 and containing the beta-chain sequence shown in SEQ ID NO: 40. The term encompasses unprocessed full-length C5 as well as any form of C5 resulting from processing in the cell. The term also encompasses naturally occurring variants of C5, for example, splicing variants or allelic variants. The amino acid sequence of an exemplary human C5 is shown in SEQ ID NO: 39 (wild-type C5 or WT C5). The amino acid sequence of the beta-chain of an exemplary human C5 is shown in SEQ ID NO: 40.The amino acid sequences of the MG1, MG2, and MG1-MG2 domains exemplifying the beta chain of human C5 are shown in SEQ ID NO: 41, 42, and 43, respectively. The amino acid sequences of C5 exemplifying cynomolgus monkey and murine C5 are shown in SEQ ID NO: 44 and 105, respectively. Amino acid residues 1 to 19 of SEQ ID NOs: 39, 40, 43, 44, and 105 correspond to a signal sequence that is removed during processing in the cell and is thus not present in the corresponding exemplary amino acid sequence.

[00134] As used herein, treatment (and its variations here) Petition 870260079322, dated 07 / 08 / 2026, p. 44 / 523 36 / 167 matical, such as treating or treating) refers to clinical intervention in an attempt to alter the natural course of the individual being treated and may be performed as prophylaxis or during the course of a clinical pathology. Desirable effects of treatment include, but are not limited to, prevention of the occurrence or recurrence of the disease, relief of symptoms, reduction of any direct or indirect pathological consequences of the disease, prevention of metastasis, reduction of the rate of disease progression, improvement or attenuation of the disease state, and remission or improved prognosis. In some embodiments, the antibodies of the invention are used to retard the development of a disease or to reduce the progression of a disease.

[00135] The term variable region or variable domain refers to the domain of a heavy or light chain of the antibody that is involved in the binding of the antibody to the antigen. The variable domains of heavy and light chains (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved structural regions (FRs) and three hypervariable regions (HVRs). (See, for example, Kindt et al., Kuby Immunology, 6th ed., WH Freeman and Co., page 91 (2007)). A single VH or VL domain may be sufficient to confer specificity in antigen binding. Furthermore, antibodies that bind to a particular antigen can be isolated using the VH or VL domain of an antibody that binds to the antigen to screen a library of complementary VH or VL domains, respectively. See, for example, Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).

[00136] The term vector, as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid molecule to which it is attached. The term includes the vector as a self-replicating nucleic acid structure as well as a vector. Petition 870260079322, dated 07 / 08 / 2026, page 45 / 523 37 / 167 incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to here as expression vectors. II. COMPOSITIONS AND METHODS

[00137] In one aspect, the invention is based, in part, on anti-C5 antibodies and their uses. In certain embodiments, antibodies are provided that bind to C5. The antibodies of the invention are useful, for example, for the diagnosis or treatment of a complement-mediated disease or condition which involves excessive or uncontrolled activation of C5. Examples of Anti-C5 Antibodies

[00138] In one aspect, the invention provides isolated antibodies that bind to C5. In certain embodiments, the anti-C5 antibody of the present invention binds to an epitope within the beta chain of C5. In certain embodiments, the anti-C5 antibody binds to an epitope within the MG1-MG2 domain of the beta chain of C5. In certain embodiments, the anti-C5 antibody binds to an epitope within a fragment consisting of amino acids 19-180 of the beta chain of C5. In certain embodiments, the anti-C5 antibody binds to an epitope within the MG1 domain (amino acids 20-124) of the beta chain of C5. In certain embodiments, the anti-C5 antibody binds to an epitope within a fragment consisting of amino acids 33-124 of the beta chain of C5 (SEQ ID NO: 40).In another embodiment, the antibody does not bind to a fragment shorter than the fragment consisting of amino acids 33-124 of the C5 beta chain, for example, a fragment consisting of amino acids 45-124, 52-124, 33-111, 33-108, or 45-111 of the C5 beta chain (SEQ ID NO: 40).

[00139] In another aspect, the invention provides anti-C5 antibodies that exhibit pH-dependent binding characteristics. As Petition 870260079322, dated 07 / 08 / 2026, p. 46 / 523 38 / 167 used herein, the expression pH-dependent binding means that the antibody exhibits reduced binding to C5 at an acidic pH when compared to its binding at a neutral pH (for the purposes of this description, both expressions may be used interchangeably). For example, antibodies with pH-dependent binding characteristics include antibodies that bind to C5 with greater affinity at a neutral pH than at an acidic pH. In certain embodiments, the antibodies of the present invention bind to C5 with an affinity at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 times greater at a neutral pH than at an acidic pH. In some embodiments, the antibodies of the present invention bind to C5 with an affinity at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 times greater at a pH of 7.4 than at a pH of 5.8.

[00140] The affinity of an antibody for C5, for the purposes of this description, is expressed in terms of the antibody's KD. The KD of an antibody refers to the dissociation equilibrium constant of an antibody-antigen interaction. The higher the KD value for the binding of an antibody to its antigen, the weaker its binding affinity for that particular antigen. Consequently, as used herein, the expression "greater affinity at a neutral pH than at an acidic pH" (or the equivalent expression "pH-dependent binding") means that the KD for antibody binding to C5 at an acidic pH is greater than the KD for antibody binding to C5 at a neutral pH. For example, in the context of the present invention, an antibody is considered to bind to C5 with greater affinity at a neutral pH than at an acidic pH if the KD of antibody binding to C5 at an acidic pH is at least 2 times greater than the KD of antibody binding to C5 at a neutral pH. Therefore, the present invention Petition 870260079322, dated 07 / 08 / 2026, p. 47 / 523 39 / 167 The invention includes antibodies that bind to C5 at an acidic pH with a KD that is at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 times greater than the KD of the antibody binding to C5 at a neutral pH. In another embodiment, the KD value of the antibody at a neutral pH may be 10⁻⁷M, 10⁻⁸M, 10⁻⁹M, 10⁻¹⁰M, 10⁻¹¹M, 10⁻¹²M or less. In another embodiment, the KD value of the antibody at an acidic pH can be 10-9M, 10-8M, 10-7M, 10-6M or more.

[00141] In further embodiments, an antibody is considered to bind to C5 with a higher affinity at a neutral pH than at an acidic pH if the KD of antibody binding to C5 at a pH of 5.8 is at least 2 times greater than the KD of antibody binding to C5 at a pH of 7.4. In some embodiments, the supplied antibodies bind to C5 at a pH of 5.8 with a KD that is at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times greater than the KD of antibody binding to C5 at a pH of 7.4. In another embodiment, the antibody KD value at a pH of 7.4 can be 10⁻⁷M, 10⁻⁸M, 10⁻⁹M, 10⁻¹⁰M, 10⁻¹¹M, 10⁻¹²M or less. In another embodiment, the antibody KD value at a pH of 5.8 can be 10⁻⁹M, 10⁻⁸M, 10⁻⁷M, 10⁻⁶M or more.

[00142] The binding properties of an antibody to a particular antigen can also be expressed in terms of the antibody's kd. The kd of an antibody refers to the rate of dissociation constant of the antibody with respect to a particular antigen and is expressed in terms of reciprocal seconds (i.e., sec-1). An increase in the kd value means weaker binding of an antibody to its antigen. Therefore, the present invention includes antibodies that bind to C5 with a higher kd value at an acidic pH than at a neutral pH. The present invention includes antibodies that bind to C5 at an acidic pH with Petition 870260079322, dated 07 / 08 / 2026, p. 48 / 523 40 / 167 a kd that is at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times greater than the kd of antibody binding to C5 at neutral pH. In another embodiment, the kd value of the antibody at neutral pH may be 10⁻²¹ / s, 10⁻³¹ / s, 10⁻⁴¹ / s, 10⁻⁵¹ / s, 10⁻⁶¹ / s or less. In another embodiment, the antibody kd value at an acidic pH can be 10⁻³¹ / s, 10⁻²¹ / s, 10⁻¹¹ / s or higher. The invention also includes antibodies that bind to C5 with a higher kd value at a pH of 5.8 than at a pH of 7.4. The present invention includes antibodies that bind to C5 at a pH of 5.8 with a kd that is at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times higher than the kd of the antibody binding to C5 at a pH of 7.4. In another embodiment, the kd value of the antibody at a pH of 7.4 can be 10-21 / s, 10-31 / s, 10-41 / s, 10-51 / s, 10-61 / s or less.In another embodiment, the kd value of the antibody at a pH of 5.8 can be 10-31 / s, 10-21 / s, 10-11 / s or more.

[00143] In certain examples, reduced C5 binding at an acidic pH compared to its binding at a neutral pH is expressed in terms of the ratio of the KD value of antibody binding to C5 at an acidic pH to the KD value of antibody binding to C5 at a neutral pH (or vice versa). For example, an antibody may be considered to exhibit reduced C5 binding at an acidic pH compared to its binding at a neutral pH, for the purposes of the present invention, if the antibody exhibits an acid / neutral KD ratio of 2 or more. In certain exemplary embodiments, the KD ratio at pH 5.8 / pH 7.4 for an antibody of the present invention is 2 or more. In certain exemplary embodiments, the acid / neutral KD ratio for an antibody of the present invention may be 3, 5, 10, 15, 20, 25, 30, 35, Petition 870260079322, dated 07 / 08 / 2026, page 49 / 523 41 / 167 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the antibody KD value at a pH of 7.4 may be 10⁻⁷M, 10⁻⁸M, 10⁻⁹M, 10⁻¹⁰M, 10⁻¹¹M, 10⁻¹²M or less. In another embodiment, the antibody KD value at a pH of 5.8 may be 10⁻⁹M, 10⁻⁸M, 10⁻⁷M, 10⁻⁶M or more.

[00144] In certain examples, reduced C5 binding at an acidic pH compared to its binding at a neutral pH is expressed in terms of the ratio of the kd value of antibody binding to C5 at an acidic pH to the kd value of antibody binding to C5 at a neutral pH (or vice versa). For example, an antibody may be considered to exhibit reduced C5 binding at an acidic pH compared to its binding at a neutral pH, for the purposes of the present invention, if the antibody exhibits an acidic / neutral kd ratio of 2 or more. In certain exemplary embodiments, the kd ratio at a pH of 5.8 / pH of 7.4 for an antibody of the present invention is 2 or more. In certain exemplary embodiments, the acid / neutral kd ratio for an antibody of the present invention may be 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more.In another embodiment, the antibody kd value at a neutral pH can be 10⁻²¹ / s, 10⁻³¹ / s, 10⁻⁴¹ / s, 10⁻⁵¹ / s, 10⁻⁶¹ / s or less. In a further embodiment, the kd value at a pH of 7.4 can be 10⁻²¹ / s, 10⁻³¹ / s, 10⁻⁴¹ / s, 10⁻⁵¹ / s, 10⁻⁶¹ / s or less. In another embodiment, the antibody kd value at an acidic pH can be 10⁻³¹ / s, 10⁻²¹ / s, 10⁻¹¹ / s or more. In a further embodiment, the antibody kd value at a pH of 5.8 can be 10⁻³¹ / s, 10⁻²¹ / s, 10⁻¹¹ / s or more.

[00145] As used herein, the expression acidic pH means a pH from 4.0 to 6.5. The expression acidic pH includes pH values ​​of any of 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, Petition 870260079322, dated 07 / 08 / 2026, p. 50 / 523 42 / 167 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4 and 6.5. In particular, the acidic pH is 5.8.

[00146] As used herein, the expression neutral pH means a pH from 6.7 to about 10.0. The expression neutral pH includes pH values ​​of any of 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 and 10.0. In particular, neutral pH is 7.4

[00147] The KD and kd values, as expressed here, can be determined using a surface plasma resonance-based biosensor to characterize antibody-antigen interactions. (See, for example, Example 3 here). The KD and kd values ​​can be determined at 25 degrees C or 37 degrees C.

[00148] In certain embodiments, an anti-C5 antibody of the present invention binds to an epitope within the beta chain of C5 consisting of the MG1 domain (SEQ ID NO: 41). In certain embodiments, an anti-C5 antibody of the present invention binds to an epitope within the beta chain (SEQ ID NO: 40) of C5 comprising at least one fragment selected from the group consisting of amino acids 47-57, 70-76 and 107-110. In certain embodiments, an anti-C5 antibody of the present invention binds to an epitope within a beta-chain fragment (SEQ ID NO: 40) of C5 comprising at least one amino acid selected from the group consisting of Thr47, Glu48, Ala49, Phe50, Asp51, Ala52, Thr53, Lys57, His70, Val71, His72, Ser74, Glu76, Val107, Ser108, ​​Lys109 and His110.In certain embodiments, an anti-C5 antibody of the present invention binds to an epitope within a beta-chain fragment (SEQ ID NO: 40) of C5 comprising at least one amino acid selected from the group consisting of Glu48. Petition 870260079322, dated 07 / 08 / 2026, p. 51 / 523 43 / 167 In certain embodiments, the binding of an anti-C5 antibody of the present invention to a C5 mutant is reduced, compared to its binding to wild-type C5, wherein the C5 mutant has at least one amino acid substitution at a position selected from the group consisting of Glu48, Asp51, His72, and Lys109. In another embodiment, the pH-dependent binding of an anti-C5 antibody of the present invention to a C5 mutant is reduced, compared to its pH-dependent binding to wild-type C5, wherein the C5 mutant has at least one amino acid substitution at a position selected from the group consisting of His70, His72, and His110. In an additional embodiment, a selected amino acid at a position between Glu48, Asp51, and Lys109 is replaced by alanine, and a selected amino acid at a position between His70, His72, and His110 is replaced by tyrosine in the C5 mutant.

[00149] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID NO: 1 and a VL of SEQ ID NO: 11; (b) a VH of SEQ ID NO: 22 and a VL of SEQ ID NO: 26; (c) a VH of SEQ ID NO: 21 and a VL of SEQ ID NO: 25; (d) a VH of SEQ ID NO: 5 and a VL of SEQ ID NO: 15; (e) a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 14; (f) a VH of SEQ ID NO: 6 and a VL of SEQ ID NO: 16; (g) a VH of SEQ ID NO: 2 and a VL of SEQ ID NO: 12; (h) a VH with SEQ ID NO: 3 and a VL with SEQ ID NO: 13; (i) a VH with SEQ ID NO: 9 and a VL with SEQ ID NO: 19; and (j) a VH with SEQ ID NO: 7 and a VL with SEQ ID NO: 17; (k) a VH with SEQ ID NO: 8 and a VL with SEQ ID NO: 18; (l) a VH with SEQ ID NO: 23 and a VL with SEQ ID NO: 27; and (m) a VH with SEQ ID NO: 10 and a VL with SEQ ID NO: 20. Petition 870260079322, dated 07 / 08 / 2026, page 52 / 523 44 / 167

[00150] In certain embodiments, an anti-C5 antibody of the present invention binds to and contacts the amino acid Asp51 (D51) of SEQ ID NO: 39. In further embodiments, an anti-C5 antibody of the present invention binds to and contacts the amino acid Lys109 (K109) of SEQ ID NO: 39. In a further embodiment, an anti-C5 antibody of the present invention binds to and contacts the amino acid Asp51 (D51) and the amino acid Lys109 (K109) of SEQ ID NO: 39.

[00151] In certain embodiments, the binding of an anti-C5 antibody of the present invention to a C5 mutant is reduced compared to its binding to wild-type C5, wherein the C5 mutant has a Glu48Ala (E48A) substitution of SEQ ID NO: 39. In another embodiment, the pH-dependent binding of an anti-C5 antibody of the present invention to a C5 mutant is reduced compared to its pH-dependent binding to wild-type C5, wherein the C5 mutant has a Glu48Ala (E48A) substitution of SEQ ID NO: 39.

[00152] In a further embodiment, an anti-C5 antibody binds to a C5 protein consisting of the amino acid sequence of SEQ ID NO: 39, but does not bind to a C5 protein consisting of the amino acid sequence of SEQ ID NO: 39 with an H72Y substitution, wherein the C5 protein and the C5 protein with the H72Y substitution are prepared and screened. under the same conditions.In a further embodiment, an anti-C5 antibody binds to a C5 protein consisting of the amino acid sequence SEQ ID NO: 39 at a pH of 7.4, but does not bind to the C5 protein with the H72Y substitution at a pH of 7.4.

[00153] Without being restricted to any particular theory, one can speculate that the binding of an anti-C5 antibody to C5 is reduced (or almost lost) when an amino acid residue on C5 is replaced by another amino acid, meaning that the amino acid residue on C5 is critical for the interactions between the anti-C5 antibody and the Petition 870260079322, dated 07 / 08 / 2026, p. 53 / 523 45 / 167 C5 and that the antibody can recognize an epitope around the amino acid residue on C5.

[00154] In the present invention, it has been discovered that a group of anti-C5 antibodies that compete with each other or bind to the same epitope can exhibit pH-dependent binding characteristics. Among the amino acids, histidine, with a pKa value of approximately 6.0 to 6.5, can have different proton dissociation states between neutral and acidic pHs. Therefore, a histidine residue on C5 can contribute to the pH-dependent interactions between an anti-C5 antibody and C5. Without being restricted to a particular theory, it can be speculated that an anti-C5 antibody may recognize a conformational structure around a histidine residue on C5 that is pH-variable.This speculation may be consistent with the experimental results described below: that the pH dependence of an anti-C5 antibody is reduced (or almost lost) when a histidine residue on C5 is replaced by another amino acid (i.e., an anti-C5 antibody with pH-dependent binding characteristics binds to a C5 histidine mutant with similar affinity to wild-type C5 at a neutral pH, while the same antibody binds to the C5 histidine mutant with a higher affinity than wild-type C5 at an acidic pH).

[00155] In certain embodiments, an anti-C5 antibody of the present invention binds to C5 from more than one species. In further embodiments, an anti-C5 antibody binds to C5 from a human and a non-human animal. In further embodiments, an anti-C5 antibody binds to C5 from a human and a monkey (e.g., cynomolgus, rhesus monkey, marmoset, chimpanzee, or baboon).

[00156] In one aspect, the invention provides anti-C5 antibodies that inhibit C5 activation. In certain embodiments, the antibodies Petition 870260079322, dated 07 / 08 / 2026, page 54 / 523 The anti-C5 antibodies provided prevent the cleavage of C5 to form C5a and C5b, thereby preventing the generation of anaphylactic activity associated with C5a, as well as preventing the assembly of the C5b-9 membrane attack complex (MAC) associated with C5b. In certain embodiments, the anti-C5 antibodies provided block the conversion of C5 to C5a and C5b by C5 convertase. In certain embodiments, the anti-C5 antibodies provided block access by C5 convertase to the cleavage site on C5. In certain embodiments, the anti-C5 antibodies provided block the hemolytic activity caused by C5 activation. In further embodiments, the anti-C5 antibodies of the present invention inhibit C5 activation via the classical and / or alternative pathways.

[00157] In one aspect, the invention provides anti-C5 antibodies that inhibit the activation of a C5 variant. A C5 variant means a genetic variant of C5 which arises due to genetic variation, such as a mutation, polymorphism, or allelic variation. A genetic variation may comprise a deletion, substitution, or insertion of one or more nucleotides. A C5 variant may comprise one or more genetic variations in C5. In certain embodiments, the C5 variant has biological activity similar to wild-type C5. Such a C5 variant may comprise at least one variation selected from the group consisting of V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N, and E1437D. Herein, R885H, for example, means a genetic variation where arginine at position 885 is replaced by histidine.In certain embodiments, an anti-C5 antibody of the present invention inhibits the activation of both wild-type C5 and at least one C5 variant selected from the group consisting of V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N, and E1437D.

[00158] In one aspect, the invention provides an anti-C5 antibody. Petition 870260079322, dated 07 / 08 / 2026, p. 55 / 523 47 / 167 comprising at least one, two, three, four, five, or six hypervariable regions (HVRs) selected from (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45-54; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55-64; (c) an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 65-74; (d) an HVR-L1 comprising the amino acid sequence of any of the SEQ ID NOs: 75-84; (e) an HVR-L2 comprising the amino acid sequence of any of the SEQ ID NOs: 85-94; and (f) an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95-104.

[00159] In one aspect, the invention provides an antibody comprising at least one, at least two, or all three HVR sequences of HV selected from (a) an HVR-H1 comprising the amino acid sequence of any of SEQ ID NOs: 45-54; (b) an HVR-H2 comprising the amino acid sequence of any of SEQ ID NOs: 55-64; and (c) an HVR-H3 comprising the amino acid sequence of any of SEQ ID NOs: 65-74. In one embodiment, the antibody comprises an HVR-H3 comprising the amino acid sequence of any of SEQ ID NOs: 65-74. In another embodiment, the antibody comprises an HVR-H3 comprising the amino acid sequence of any of SEQ ID NOs: 65-74 and an HVR-L3 comprising the amino acid sequence of any of SEQ ID NOs: 95-104.In a further embodiment, the antibody comprises an HVR-H3 comprising the amino acid sequence of any of SEQ ID NOs: 65-74, an HVR-L3 comprising the amino acid sequence of any of SEQ ID NOs: 95-104, and an HVR-H2 comprising the amino acid sequence of which... Petition 870260079322, dated 07 / 08 / 2026, p. 56 / 523 48 / 167 requires one of SEQ ID NOs: 55-64. In a further embodiment, the antibody comprises (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45-54; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55-64; and (c) an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 65-74.

[00160] In another aspect, the invention provides an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) an HVR-L1 comprising the amino acid sequence of any of SEQ ID NOs: 75-84; (b) an HVR-L2 comprising the amino acid sequence of any of SEQ ID NOs: 85-94; and (c) an HVR-L3 comprising the amino acid sequence of any of SEQ ID NOs: 95-104. In one embodiment, the antibody comprises (a) an HVR-L1 comprising the amino acid sequence of any of SEQ ID NOs: 75-84; (b) an HVR-L2 comprising the amino acid sequence of any of SEQ ID NOs: 85-94; and (c) an HVR-L3 comprising the amino acid sequence of any of SEQ ID NOs: 95-104.

[00161] In another aspect, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (i) an HVR-H1 comprising the amino acid sequence of any of SEQ ID NOs: 45-54, (ii) an HVR-H2 comprising the amino acid sequence of any of SEQ ID NOs: 55-64, and (iii) an HVR-H3 comprising the amino acid sequence of any of SEQ ID NOs: 65-74; and (b) a VL domain comprising at least one, at least two, or all three VL HVR sequences selected from (i) an HVR-L1 comprising the amino acid sequence of any of SEQ ID NOs: Petition 870260079322, dated 07 / 08 / 2026, p. 57 / 523 49 / 167 75-84, (ii) an HVR-L2 comprising the amino acid sequence of any of the SEQ ID NOs: 85-94 and (c) an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95-104.

[00162] In another aspect, the invention provides an antibody comprising (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45-54; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55-64; (c) an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 65-74; (d) an HVRL1 comprising the amino acid sequence of any of the SEQ ID NOs: 75-84; (e) an HVR-L2 comprising the amino acid sequence of any of the SEQ ID NOs: 85-94; and (f) an HVRL3 comprising the amino acid sequence of any of the SEQ ID NOs: 95-104.

[00163] In one aspect, the invention provides an anti-C5 antibody comprising at least one, two, three, four, five or six HVRs selected from (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45, 54, 117, 126; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55, 64, 118-120, 127; (c) an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 65, 74, 121, 128; (d) an HVR-L1 comprising the amino acid sequence of any of the SEQ ID NOs: 75, 84, 122, 129; (e) an HVR-L2 comprising the amino acid sequence of any of the SEQ ID NOs: 85, 94, 123-124, 130; and (f) an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95, 104, 125, 131.

[00164] In one aspect, the invention provides an antibody comprising at least one, at least two, or all three sequences Petition 870260079322, dated 07 / 08 / 2026, p. 58 / 523 50 / 167 HVR cies of VH selected from (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45, 54, 117, 126; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55, 64, 118-120, 127; (c) an HVR-H3 comprising the amino acid sequence of any one of SEQ ID NOs: 65, 74, 121, 128. In one embodiment, the antibody comprises an HVR-H3 comprising the amino acid sequence of any one of SEQ ID NOs: 65, 74, 121, 128. In another embodiment, the antibody comprises an HVR-H3 comprising the amino acid sequence of any one of SEQ ID NOs: 65, 74, 121, 128 and an HVR-L3 comprising the amino acid sequence of any one of SEQ ID NOs: 95, 104, 125, 131.In a further embodiment, the antibody comprises an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 65, 74, 121, 128, an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95, 104, 125, 131 and an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55, 64, 118-120, 127. In a further embodiment, the antibody comprises (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45, 54, 117, 126; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55, 64, 118-120, 127; and (c) an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 65, 74, 121, 128.

[00165] In another aspect, the invention provides an antibody comprising at least one, at least two, or all three HVR sequences of VL selected from (a) an HVR-L1 comprising the amino acid sequence of any of the SEQ ID NOs: 75, 84, 122, 129; (b) an HVR-L2 comprising the amino acid sequence of any of the SEQ ID NOs: 85, 94, 123-124, Petition 870260079322, dated 07 / 08 / 2026, p. 59 / 523 51 / 167 130; and (c) an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95, 104, 125, 131. In one embodiment, the antibody comprises (a) an HVR-L1 comprising the amino acid sequence of any of the SEQ ID NOs: 75, 84, 122, 129; (b) an HVR-L2 comprising the amino acid sequence of any of the SEQ ID NOs: 85, 94, 123-124, 130; and (c) an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95, 104, 125, 131.

[00166] In another aspect, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two or all three VH HVR sequences selected from (i) an HVR-H1 comprising the amino acid sequence of any of SEQ ID NOs: 45, 54, 117, 126, (ii) an HVR-H2 comprising the amino acid sequence of any of SEQ ID NOs: 55, 64, 118-120, 127 and (iii) an HVR-H3 comprising the amino acid sequence of any of SEQ ID NOs: 65, 74, 121, 128; and (b) a VL domain comprising at least one, at least two, or all three HVR VL sequences selected from (i) an HVR-L1 comprising the amino acid sequence of any of the SEQ ID NOs: 75, 84, 122, 129, (ii) an HVRL2 comprising the amino acid sequence of any of the SEQ ID NOs: 85, 94, 123-124, 130, and (c) an HVR-L3 comprising the amino acid sequence of any of the SEQ ID NOs: 95, 104, 125, 131.

[00167] In another aspect, the invention provides an antibody comprising (a) an HVR-H1 comprising the amino acid sequence of any of the SEQ ID NOs: 45, 54, 117, 126; (b) an HVR-H2 comprising the amino acid sequence of any of the SEQ ID NOs: 55, 64, 118-120, 127; (c) an HVR-H3 comprising the amino acid sequence of any of the SEQ ID NOs: 55, 64, 118-120, 127; Petition 870260079322, dated 07 / 08 / 2026, p. 60 / 523 52 / 167 (d) an HVR-L1 comprising the amino acid sequence of any of the SEQ IDs: 75, 84, 122, 129; (e) an HVR-L2 comprising the amino acid sequence of any of the SEQ IDs: 85, 94, 123-124, 130; and (f) an HVRL3 comprising the amino acid sequence of any of the SEQ IDs: 95, 104, 125, 131.

[00168] In certain embodiments, any one or more of the amino acids of an anti-C5 antibody as provided above are substituted at the following HVR positions: (a) in HVR-H1 (SEQ ID NO: 45), at positions 5 and 6; (b) in HVR-H2 (SEQ ID NO: 55), at positions 1, 3, 9, 11, 13 and 15; (c) in HVR-H3 (SEQ ID NO: 65), at positions 2, 5, 6, 12 and 13; (d) in HVR-L1 (SEQ ID NO: 75), at positions 1, 5, 7 and 9; (e) in HVR-L2 (SEQ ID NO: 85), at positions 4, 5 and 6; and (f) in HVR-L3 (SEQ ID NO: 95), at positions 2, 4 and 12.

[00169] In certain embodiments, the substitutions are conservative substitutions, as provided herein. In certain embodiments, any one or more of the following substitutions may be made in any combination: (a) in HVR-H1 (SEQ ID NO: 45), M5V or C6A; (b) in HVR-H2 (SEQ ID NO: 55), C1A or G, Y3F, T9D or E, Y11K or Q, S13D or E or A15V; (c) in HVR-H3 (SEQ ID NO: 65), G2A, V5Q or D, T6Y, Y12H or L13Y; (d) in HVR-L1 (SEQ ID NO: 75), Q1R, N5Q or G, G7S, D9K or S; (e) in HVR-L2 (SEQ ID NO: 85), K4T or E, L5T or A6H, A6 E or A6Q; (f) in HVR-L3 (SEQ ID NO: 95) C2S, C2N or C2T, F4K; or A12T or A12H.

[00170] All possible combinations of the above substitutions are covered by the consensus sequences with SEQ ID numbers: 126, 127, 128, 129, 130 and 131 for HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3, respectively.

[00171] In any of the above embodiments, an anti-C5 antibody is humanized. In one embodiment, an anti-C5 antibody Petition 870260079322, dated 07 / 08 / 2026, p. 61 / 523 53 / 167 comprises HVRs according to any of the above embodiments and additionally comprises a human acceptor structural region, for example, a human immunoglobulin structural region or a human consensus structural region. In another embodiment, an anti-C5 antibody comprises HVRs according to any of the above embodiments and additionally comprises a VH or VL comprising an FR sequence, wherein the FR sequences are as follows. For the heavy chain variable domain, FR1 comprises the amino acid sequence of any of the SEQ ID NOs: 132-134, FR2 comprises the amino acid sequence of any of the SEQ ID NOs: 135-136, FR3 comprises the amino acid sequence of any of the SEQ ID NOs: 137-139, FR4 comprises the amino acid sequence of any of the SEQ ID NOs: 140-141.For the variable light chain domain, FR1 comprises the amino acid sequence of any of the SEQ ID NOs: 142-143, FR2 comprises the amino acid sequence of any of the SEQ ID NOs: 144-145, FR3 comprises the amino acid sequence of any of the SEQ ID NOs: 146-147, FR4 comprises the amino acid sequence of SEQ ID NO: 148.

[00172] In another aspect, an anti-C5 antibody comprises a variable heavy chain (VH) domain sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of any of SEQ ID NOs: 1-10. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids Petition 870260079322, dated 07 / 08 / 2026, p. 62 / 523 54 / 167 acids were substituted, inserted, and / or deleted in any of SEQ ID NOs: 1-10. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs). Optionally, the anti-C5 antibody comprises the HVR sequence in any of the SEQ ID NOs: 1-10, including post-translational modifications of this sequence. In one particular embodiment, the HVR comprises one, two, or three HVRs selected from: (a) HVR-H1 comprising an amino acid sequence from any of the SEQ ID NOs: 45-54, (b) HVR-H2 comprising an amino acid sequence from any of the SEQ ID NOs: 55-64, and (c) HVR-H3 comprising an amino acid sequence from any of the SEQ ID NOs: 65-74.

[00173] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a variable light chain (VL) domain that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of any of SEQ ID NOs: 11-20. In certain embodiments, a VL sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in any of the SEQ ID NOs: 11-20. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in FRs).Optionally, the anti-C5 antibody comprises the VL sequence in any of the SEQ ID NOS: 11-20, including post-translational modifications of this sequence. In one particular embodiment, the VL comprises one, two, or three of the HVRs. Petition 870260079322, dated 07 / 08 / 2026, p. 63 / 523 55 / 167 administered of (a) HVR-L1 comprising an amino acid sequence of any of the SEQ ID NOs: 75-84; (b) HVR-L2 comprising an amino acid sequence of any of the SEQ ID NOs: 85-94; and (c) HVR-L3 comprising an amino acid sequence of any of the SEQ ID NOs: 95-104.

[00174] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a VH according to any of the embodiments provided above and a VL according to any of the embodiments provided above. In one embodiment, the antibody comprises the VH and VL sequences in any of SEQ ID NOs: 1-10 and any of SEQ ID NOs: 11-20, respectively, including post-translational modifications of these sequences.

[00175] In another aspect, an anti-C5 antibody comprises a variable heavy chain (VH) domain sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of any of the SEQ ID NOs: 10, 106-110. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in any of the SEQ ID NOs: 10, 106-110. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs).Optionally, the anti-C5 antibody comprises the VH sequence in any of the SEQ ID NOs: 10, 106110, including post-translational modifications of this sequence. In one particular embodiment, VH comprises one, two, or three of the following. Petition 870260079322, dated 07 / 08 / 2026, p. 64 / 523 56 / 167 Selected HVRs from: (a) HVR-H1 comprising an amino acid sequence from any of the SEQ ID NOs: 45, 54, 117, 126, (b) HVR-H2 comprising an amino acid sequence from any of the SEQ ID NOs: 55, 64, 118-120, 127 and (c) HVR-H3 comprising an amino acid sequence from any of the SEQ ID NOs: 65, 74, 121, 128.

[00176] In another aspect, an anti-C5 antibody comprises a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of any of the SEQ ID NOS: 10, 106-110. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in any of the SEQ ID NOs: 10, 106-110. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs).Optionally, the anti-C5 antibody comprises the VH sequence in any of the SEQ ID NOs: 10, 106-110, including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two, or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence from any of the SEQ ID NOs: 45, 54, 117, 126, (b) HVR-H2 comprising an amino acid sequence from any of the SEQ ID NOs: 55, 64, 118-120, 127, and (c) HVR-H3 comprising an amino acid sequence from any of the SEQ ID NOs: 65, 74, 121, 128.

[00177] In another aspect, an anti-C5 antibody comprises a Petition 870260079322, dated 07 / 08 / 2026, p. 65 / 523 57 / 167 VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the VH sequence is the amino acid sequence of SEQ ID NO: 10. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids were substituted, inserted and / or deleted in SEQ ID NO: 10. In certain embodiments, substitutions, insertions or deletions occur outside the HVR regions (i.e., in the FRs).Optionally, the anti-C5 antibody comprises the VH sequence at SEQ ID NO: 10 including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two, or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence at SEQ ID NO: 54, (b) HVR-H2 comprising an amino acid sequence at SEQ ID NO: 64, and (c) HVR-H3 comprising an amino acid sequence at SEQ ID NO: 74.

[00178] In another aspect, an anti-C5 antibody comprises a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 106. In certain embodiments, the VH sequence is an amino acid sequence of SEQ ID NO: 106. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions in Petition 870260079322, dated 07 / 08 / 2026, p. 66 / 523 58 / 167 relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in SEQ ID NO: 106. In certain embodiments, substitutions, insertions or deletions occur outside the HVR regions (i.e., in the FRs). Optionally, the anti-C5 antibody comprises the VH sequence in SEQ ID NO: 106, including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence from SEQ ID NO: 117, (b) HVR-H2 comprising an amino acid sequence from SEQ ID NO: 118 and (c) HVR-H3 comprising an amino acid sequence from SEQ ID NO: 121.

[00179] In another aspect, an anti-C5 antibody comprises a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 107. In certain embodiments, the VH sequence is an amino acid sequence of SEQ ID NO: 107. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody The anti-C5 sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 107. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs).Optionally, the anti-C5 antibody comprises the VH sequence in SEQ ID NO: 107. Petition 870260079322, dated 07 / 08 / 2026, p. 67 / 523 59 / 167 including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two, or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence of SEQ ID NO: 117 (b) HVR-H2 comprising an amino acid sequence of SEQ ID NO: 119 and (c) HVRH3 comprising an amino acid sequence of SEQ ID NO: 121.

[00180] In another aspect, an anti-C5 antibody comprises a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 108. In certain embodiments, the VH sequence is an amino acid sequence of SEQ ID NO: 108. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody The anti-C5 sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 108. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs).Optionally, the anti-C5 antibody comprises the VH sequence at SEQ ID NO: 108, including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two, or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence at SEQ ID NO: 117, (b) HVR-H2 comprising an amino acid sequence at SEQ ID NO: 118, and (c) HVR-H3 comprising an amino acid sequence at SEQ ID NO: 121. Petition 870260079322, dated 07 / 08 / 2026, p. 68 / 523 60 / 167

[00181] In another aspect, an anti-C5 antibody comprises a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 109. In certain embodiments, the VH sequence is an amino acid sequence of SEQ ID NO: 109. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody The anti-C5 sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 109. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs).Optionally, the anti-C5 antibody comprises the VH sequence at SEQ ID NO: 109, including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two, or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence at SEQ ID NO: 117, (b) HVR-H2 comprising an amino acid sequence at SEQ ID NO: 118, and (c) HVR-H3 comprising an amino acid sequence at SEQ ID NO: 121.

[00182] In another aspect, an anti-C5 antibody comprises a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 110. In certain embodiments, the VH sequence is an amino acid sequence of SEQ ID NO: 110. In certain embodiments, a VH sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, Petition 870260079322, dated 07 / 08 / 2026, p. 69 / 523 61 / 167%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 110. In certain embodiments, substitutions, insertions, or deletions occur outside the HVR regions (i.e., in the FRs). Optionally, the anti-C5 antibody comprises the VH sequence in SEQ ID NO: 110, including post-translational modifications of this sequence. In a particular embodiment, the VH comprises one, two, or three of the HVRs selected from: (a) HVR-H1 comprising an amino acid sequence with SEQ ID NO: 117, (b) HVR-H2 comprising an amino acid sequence with SEQ ID NO: 120, and (c) HVRH3 comprising an amino acid sequence with SEQ ID NO: 121.

[00183] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a variable light chain (VL) domain that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of any of SEQ ID NOs: 20, 111-113. In certain embodiments, a VL sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in any of the SEQ ID NOs: 20, 111-113. In certain embodiments, substitutions, insertions, or deletions occur outside Petition 870260079322, dated 07 / 08 / 2026, p. 70 / 523 62 / 167 of the HVR regions (i.e., in the FRs). Optionally, the anti-C5 antibody comprises the VL sequence in any of the SEQ ID NOs: 20, 111-113, including post-translational modifications of this sequence. In a particular embodiment, the VL comprises one, two, or three of the HVRs selected from (a) HVR-L1 comprising an amino acid sequence from any of the SEQ ID NOs: 75, 84, 122, 129; (b) HVR-L2 comprising an amino acid sequence from any of the SEQ ID NOs: 85, 94, 123-124, 130; and (c) HVR-L3 comprising an amino acid sequence from any of the SEQ ID NOs: 95, 104, 125, 131.

[00184] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a VL that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 20. In certain embodiments, the VL sequence is an amino acid sequence of SEQ ID NO: 20. In certain embodiments, a VL sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but an anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids were substituted, inserted and / or deleted in SEQ ID NO: 20. In certain embodiments, substitutions, insertions or deletions occur outside the HVR regions (i.e., in FRs).Optionally, the anti-C5 antibody comprises the VL sequence at SEQ ID NO: 20, including post-translational modifications of this sequence. In a particular embodiment, the VL comprises one, two, or three of the HVRs selected from (a) HVR-L1 comprising an amino acid sequence at SEQ ID NO: 84; (b) HVR-L2 comprising... Petition 870260079322, dated 07 / 08 / 2026, p. 71 / 523 63 / 167 comprises an amino acid sequence with SEQ ID NO: 94; and (c) HVR-L3 comprises an amino acid sequence with SEQ ID NO: 104.

[00185] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a VL that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 111. In certain embodiments, the VL sequence is an amino acid sequence of SEQ ID NO: 111. In certain embodiments, a VL sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but An anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 111. In certain embodiments, substitutions, insertions, or deletions occur outside of HVR regions (i.e., in FRs).Optionally, the anti-C5 antibody comprises the VL sequence at SEQ ID NO: 111, including post-translational modifications of this sequence. In a particular embodiment, the VL comprises one, two, or three of the HVRs selected from (a) HVR-L1 comprising an amino acid sequence at SEQ ID NO: 122; (b) HVR-L2 comprising an amino acid sequence at SEQ ID NO: 123; and (c) HVR-L3 comprising an amino acid sequence at SEQ ID NO: 125.

[00186] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a VL that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: Petition 870260079322, dated 07 / 08 / 2026, p. 72 / 523 64 / 167 112. In certain embodiments, the VL sequence is the amino acid sequence of SEQ ID NO: 112. In certain embodiments, the VL sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody comprising that sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 112. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-C5 antibody comprises the VL sequence at SEQ ID NO: 112, including post-translational modifications of this sequence.In a particular embodiment, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence with SEQ ID NO: 122; (b) HVR-L2 comprising the amino acid sequence with SEQ ID NO: 123; and (c) HVR-L3 comprising the amino acid sequence with SEQ ID NO: 125.

[00187] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a VL that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 113. In certain embodiments, the VL sequence is the amino acid sequence of SEQ ID NO: 113. In certain embodiments, the VL sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but An anti-C5 antibody comprising this sequence retains the ability to bind to C5. In certain embodiments, a total of 1 to 10 amino acids were... Petition 870260079322, dated 07 / 08 / 2026, p. 73 / 523 65 / 167 substituted, inserted, and / or deleted in SEQ ID NO: 113. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-C5 antibody comprises the VL sequence in SEQ ID NO: 113, including post-translational modifications of this sequence. In a particular embodiment, the VL comprises one, two, or three HVRs selected from (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 124; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125.

[00188] In another aspect, an anti-C5 antibody is provided, wherein the antibody comprises a VH according to any of the embodiments provided above and a VL according to any of the embodiments provided above. In one embodiment, the antibody comprises VH and VL sequences in any of SEQ ID NOs: 10, 106-110 and in any of SEQ ID NOs: 20, 111-113, respectively, including post-translational modifications of these sequences. In one embodiment, the antibody comprises a VH sequence from SEQ ID NO: 10 and a VL sequence from SEQ ID NO: 20. In one embodiment, the antibody comprises a VH sequence from SEQ ID NO: 106 and a VL sequence from SEQ ID NO: 111.

[00189] In another embodiment, the antibody comprises a VH sequence from SEQ ID NO: 107 and a VL sequence from SEQ ID NO: 111. In a further embodiment, the antibody comprises a VH sequence from SEQ ID NO: 108 and a VL sequence from SEQ ID NO: 111. In another embodiment, the antibody comprises a VH sequence from SEQ ID NO: 109 and a VL sequence from SEQ ID NO: 111. In another embodiment, the antibody comprises a sequence Petition 870260079322, dated 07 / 08 / 2026, p. 74 / 523 66 / 167 VH sequence of SEQ ID NO: 109 and a VL sequence of SEQ ID NO: 112. In another embodiment, the antibody comprises a VH sequence of SEQ ID NO: 109 and a VL sequence of SEQ ID NO: 113. In another embodiment, the antibody comprises a VH sequence of SEQ ID NO: 110 and a VL sequence of SEQ ID NO: 113.

[00190] In one aspect, an anti-C5 antibody is provided wherein the antibody comprises a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 54, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 64, and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 74 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 84; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 94; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 104.

[00191] In another aspect, an anti-C5 antibody is provided wherein the antibody comprises a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118, and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 123; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125.

[00192] In another aspect, an anti-C5 antibody is provided wherein the antibody comprises a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 119 and (c) an HVR-H3 comprising the sequence Petition 870260079322, dated 07 / 08 / 2026, p. 75 / 523 67 / 167 amino acids of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 123; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125.

[00193] In another aspect, an anti-C5 antibody is provided wherein the antibody comprises a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118, and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 124; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125.

[00194] In certain embodiments, an anti-C5 antibody of the present invention comprises a VH according to any of the embodiments provided above and a heavy chain constant region comprising the amino acid sequence of any of the SEQ ID NOs: 33, 34, 35, 114, 115 and 116. In certain embodiments, an anti-C5 antibody of the present invention comprises a VL according to any of the embodiments provided above and a light chain constant region comprising the amino acid sequence of any of the SEQ ID NOs: 36, 37 and 38.

[00195] In another aspect, the invention provides an antibody that binds to the same epitope as an anti-C5 antibody provided herein. For example, in certain embodiments, an antibody is provided that binds to the same epitope as an antibody described in Table 2. As demonstrated by the working examples below, all anti Petition 870260079322, dated 07 / 08 / 2026, p. 76 / 523 68 / 167 anti-C5 bodies described in Table 2 are grouped in the same compartment as the C5 epitope and exhibit pH-dependent binding characteristics.

[00196] In a further aspect, the invention provides an antibody that binds to the same epitope as an antibody provided herein. In a further aspect, the invention provides an antibody that binds to the same epitope as an antibody described in Tables 7 or 8. In certain embodiments, an antibody is provided that binds to an epitope within a fragment consisting of amino acids 33 to 124 of the beta chain of C5 (SEQ ID NO: 40), comprising at least one fragment selected from the group consisting of amino acids 47-57, 70-76 and 107-110. In certain embodiments, an antibody is provided that binds to an epitope within a fragment consisting of amino acids 33 to 124 of the beta chain of C5 (SEQ ID NO: 40) comprising at least one fragment selected from the group consisting of amino acids Thr47, Glu48, Ala49, Phe50, Asp51, Ala52, Thr53, Lys57, His70, Val71, His72, Ser74, Glu76, Val107, Ser108, ​​Lys109 and His110.In another embodiment, an epitope of an anti-C5 antibody of the present invention is a conformational epitope.

[00197] In a further aspect of the invention, an anti-C5 antibody according to any of the above embodiments is a monoclonal antibody, including a chimeric, humanized or human antibody. In one embodiment, an anti-C5 antibody is an antibody fragment, for example, an Fv, Fab, Fab', scFv, diabody or F(ab')2 fragment. In another embodiment, the antibody is a full-length antibody, for example, an intact IgG1 or IgG4 antibody or another antibody class or isotype as defined herein.

[00198] In a further aspect, an anti-C5 antibody according to any of the above embodiments may incorporate any Petition 870260079322, dated 07 / 08 / 2026, p. 77 / 523 69 / 167 one of the features, alone or in combination, as described in Sections 1-7 below. Antibody Affinity

[00199] In certain embodiments, an antibody provided here has a dissociation constant (Kd) of 1 micro M or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less or 0.001 nM or less (e.g., 10-8M or less, e.g., from 10-8M to 10-13M, e.g., from 10-9M to 10-13M).

[00200] In one embodiment, Kd is measured by means of a radiolabeled antigen binding assay (RIA). In one embodiment, an RIA is performed with a Fab version of an antibody of interest and its antigen. For example, the affinity of a Fab binding solution for an antigen is measured by equilibrating Fab with a minimum concentration of (125I)-labeled antigen in the presence of a titration series of unlabeled antigen, then capturing the bound antigen with a plate coated with an anti-Fab antibody (see, for example, Chen et al., J. Mol. Biol. 293: 865-881(1999)).To establish the conditions for the assay, multi-well MICROTITER plates (registered trademark) (Thermo Scientific) are coated overnight with 5 µg / ml of an anti-Fab capture antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6) and subsequently blocked with 2% (w / v) bovine serum albumin in PBS for two to five hours at room temperature (approximately 23°C). In a non-adsorbent plate (Nunc #269620), 100 pM or 26 pM [125I]-antigen is mixed with serial dilutions of a Fab of interest (e.g., consistent with the evaluation of the anti-VEGF antibody, Fab-12, in Presta et al., Cancer Res. 57: 4593-4599 (1997)). The Fab of interest is then incubated overnight; however, incubation may continue for one day. Petition 870260079322, dated 07 / 08 / 2026, page 78 / 523 70 / 167 longer period (e.g., approximately 65 hours) to ensure equilibrium is reached. Then, the mixtures are transferred to the capture plate for incubation at room temperature (e.g., for one hour). The solution is then removed and the plate washed eight times with 0.1% polysorbate 20 (TWEEN-20 (trademark)) in PBS. When the plates are dry, 150 µl / well of scintillating agent (MICROSCINT-20™; Packard) are added and the plates are counted on a TOPCOUNT™ gamma counter (Packard) for ten minutes. Concentrations of each Fab that give less than or equal to 20% of the maximum binding are chosen for use in competitive binding assays.

[00201] According to another embodiment, Kd is measured using a BIACORE (trademark) surface plasma resonance assay. For example, an assay using BIACORE (trademark) 2000 or BIACORE (trademark) 3000 (BIACORE (trademark), Inc., Piscataway, NJ) is performed at 25 degrees C with the antigen immobilized on CM5 chips at ~10 response units (RU). In one embodiment, carboxymethylated dextran biosensor chips (CM5, BIACORE (trademark), Inc.) are activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodi-imide hydrochloride (EDC) and N-hydroxy-succinimide (NHS) according to the vendor's instructions. The antigen is diluted with 10 mM sodium acetate, pH 4.8, to 5 µg / ml (~0.2 µM) before injection at a flow rate of 5 µL / minute to obtain approximately 10 response units (RU) of the bound protein. After antigen injection, 1 M ethanolamine is injected to block unreacted groups.For kinetic measurements, serial twice dilutions of Fab (0.78 nM to 500 nM) are injected into PBS with 0.05% polysorbate 20 surfactant (TWEEN-20™) (in English, PBST) at 25 degrees C at a flow rate of approximately 25 micro l / minute. Petition 870260079322, dated 07 / 08 / 2026, page 79 / 523 71 / 167 Association rates (kon) and dissociation rates (koff) are calculated using a simple one-to-one Langmuir linkage model (BIACORE Assessment Software (registered trademark) version 3.2) by simultaneously fitting association and dissociation sensorgrams. The dissociation equilibrium constant (Kd) is calculated as the koff / kon ratio. See, for example, Chen et al., J. Mol. Biol. 293: 865-881 (1999).If the association rate constant exceeds 106M-1s-1 by the surface plasma resonance assay above, then the association rate constant can be determined using the fluorescence quenching technique which measures the increase or decrease in fluorescence emission intensity (excitation = 295 nm; emission = 340 nm, 16 nm bandpass) at 25 degrees C of a 20 nM anti-antigen antibody (Fab form) in PBS, pH 7.2, in the presence of increasing antigen concentrations, as measured in a spectrophotometer, such as a stop-flow equipped spectrophotometer (Aviv Instruments) or an 8000 series SLM-AMINCO™ spectrophotometer (ThermoSpectronic) with a stirring cuvette. 2. Antibody Fragments

[00202] In certain embodiments, an antibody provided herein is an antibody fragment. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments and other fragments described below. For a review of specific antibody fragments, see Hudson et al., Nat. Med. 9: 129-134 (2003). For a review of scFv fragments, see, for example, Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pages 269-315 (1994); see also WO 93 / 16185; and U.S. Patents Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab')2 fragments comprising residues of... Petition 870260079322, dated 07 / 08 / 2026, page 80 / 523 72 / 167 operation of the binding epitope and which have an increased in vivo half-life see U.S. Patent No. 5,869,046.

[00203] Diabodies are antibody fragments with two antigen-binding sites that can be bivalent or bispecific. See, for example, EP 404,097; WO1993 / 01161; Hudson et al., Nat. Med. 9: 129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9: 129-134 (2003).

[00204] Single-domain antibodies are antibody fragments comprising all or part of the variable heavy chain domain or all or part of the variable light chain domain of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, for example, U.S. Patent No. 6,248,516).

[00205] Antibody fragments can be made by means of various techniques including, but not limited to, proteolytic digestion of an intact antibody, as well as production by recombinant host cells (e.g., E. coli or phage), as described herein. 3. Chimeric and Humanized Antibodies

[00206] In certain embodiments, an antibody provided herein is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81: 6851-6855 (1984)). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate, such as a monkey) and a human constant region. In a further example, a chimeric antibody is a class-swapped antibody, in which the class or subclass has been swapped relative to that of the antibody. Petition 870260079322, dated 07 / 08 / 2026, page 81 / 523 73 / 167 parental. Chimeric antibodies include antigen-binding fragments of the same.

[00207] In certain embodiments, a chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. In general, a humanized antibody comprises one or more variable domains, in which HVRs, for example, CDRs (or portions thereof) are derived from a non-human antibody and FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody will optionally also comprise at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are replaced by corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), for example, to restore or enhance the antibody's specificity or affinity.

[00208] Humanized antibodies and methods for making them are reviewed in, for example, Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008) and are further described, for example, in Riechmann et al., Nature 332: 323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86: 10029-10033 (1989); United States Patents Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Kashmiri et al., Methods 36: 25-34 (2005) (which describes the complementarity-determining region (CDR) graft); Padlan, Mol. Immunol. 28: 489-498 (1991) (which describes resurfacing); Dall'Acqua et al., Methods 36: 4360 (2005) (which describes FR shuffling); and Osbourn et al., Methods 36: 61-68 (2005) and Klimka et al., Br. J. Cancer 83: 252-260 (2000) (which describes the guided selection approach for FR shuffling).

[00209] Human structural regions that can be used for the Petition 870260079322, dated 07 / 08 / 2026, p. 82 / 523 74 / 167 humanization includes, but is not limited to: structural regions selected using the best-fit method (see, for example, Sims et al., J. Immunol. 151: 2296 (1993)); structural regions derived from the consensus sequence of human antibodies from a particular subgroup of variable light or heavy chain regions (see, for example, Carter et al., Proc. Natl. Acad. Sci. USA 89: 4285 (1992); and Presta et al., J. Immunol. 151: 2623 (1993)); mature human structural regions (with somatic mutation) or human germline structural regions (see, for example, Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008)); and structural regions derived from screening FR libraries (see, for example, Baca et al., J. Biol. Chem. 272: 10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271: 22611-22618 (1996)). 4. Human Antibodies

[00210] In certain embodiments, an antibody provided herein is a human antibody. Human antibodies can be produced using various known techniques. Human antibodies are described in general in van Dijk and van de Winkel, Curr. Opin. Pharma. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

[00211] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigenic challenge. Such animals typically contain all or a portion of the human immunoglobulin loci, which replace endogenous immunoglobulin loci or are present extrachromosomally or randomly integrated into the animal's chromosomes. In such transgenic mice, the transgenic immunoglobulin loci are usually inactivated.For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg. Petition 870260079322, dated 07 / 08 / 2026, p. 83 / 523 75 / 167 Nat. Biotech. 23: 1117-1125 (2005). See also, for example, U.S. Patents Nos. 6,075,181 and 6,150,584, which describe the XENOMOUSE™ technology; U.S. Patent No. 5,770,429, which describes the HUMAB technology (trademark); U.S. Patent No. 7,041,870, which describes the KM MOUSE technology (trademark); and U.S. Patent Application Publication No. US 2007 / 0061900, which describes the VELOCIMOUSE technology (trademark)). Human variable regions of intact antibodies generated by such animals may be further modified, for example, by combining with a different human constant region.

[00212] Human antibodies can also be made using hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described (see, for example, Kozbor, J. Immunol. 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pages 5163 (Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol. 147: 86 (1991).) Human antibodies generated using human B-cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA 103: 3557-3562 (2006). Additional methods include those described, for example, in U.S. Patent No. 7,189,826 (which describes the production of human monoclonal IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue 26(4): 265-268 (2006) (which describes human-human hybridomas).Human hybridoma technology (Triome Technology) is also described in Vollmers, Histology and Histopathology 20(3): 927-937 (2005) and Vollmers, Methods and Findings in Experimental and Clinical Pharmacology 27(3): 185-191 (2005).

[00213] Human antibodies can also be generated through Petition 870260079322, dated 07 / 08 / 2026, page 84 / 523 76 / 167 Isolation of variable domain sequences from a Fv clone selected from human-derived phage display libraries. Such variable domain sequences can be combined with a desired human constant domain. Techniques for selecting human antibodies from antibody libraries are described below. 5. Library-Derived Antibodies

[00214] The antibodies of the invention can be isolated by screening combinatorial libraries for antibodies with the desired activity or activities. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing the desired binding characteristics. Such methods are reviewed, for example, in Hoogenboom et al., Methods in Molecular Biology 178: 137 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001) and further described, for example, in McCafferty et al., Nature 348: 552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks, Meth.Mol. Biol. 248: 161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Academic. Sci. USA 101(34): 12467-12472 (2004); Lee et al., J. Immunol.Methods 284(1-2): 119-132 (2004).

[00215] In certain phage display methods, VH and VL gene repertoires are separately cloned by polymerase chain reaction (PCR) and randomly recombined into phage libraries which can be screened for the antigen-binding phage, as described in Winter et al., Ann. Rev. Immunol. 12: 433-455 (1994). Phages typically present as antibody fragments or single-stranded Fv fragments (scFv) or as Fab fragments. Phage libraries Petition 870260079322, dated 07 / 08 / 2026, page 85 / 523 77 / 167 immunized cells provide high-affinity antibodies for the immunogen without the need for hybridoma construction. Alternatively, a virgin repertoire can be cloned to provide a single source of antibodies for a wide variety of non-self and self antigens without any immunization, as described in Griffiths et al., EMBO J, 12: 725-734 (1993). Finally, virgin libraries can also be made synthetically by cloning non-rearranged V gene segments from stem cells and using PCR primers containing a random sequence to encode the highly variable CDR3 regions and achieve in vitro rearrangement, as described by Hoogenboom, J. Mol. Biol. 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example: U.S. Patent No. 5,750,373 and U.S. Publications Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598. 2007 / 0237764, 2007 / 0292936 and 2009 / 0002360.

[00216] Antibodies or antibody fragments isolated from human antibody libraries are considered here as human antibodies or human antibody fragments. 6. Multispecific Antibodies

[00217] In certain embodiments, an antibody provided here is a multispecific antibody, for example, a bispecific antibody. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In certain embodiments, one of the binding specificities is for C5 and the other is for any other antigen. In certain embodiments, bispecific antibodies can bind to two different epitopes of C5. Bispecific antibodies can also be used to target cytotoxic agents to cells expressing C5. Bispecific antibodies can be prepared as antibodies to Petition 870260079322, dated 07 / 08 / 2026, page 86 / 523 78 / 167 full length or antibody fragments.

[00218] Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy-chain-light chain pairs that have different specificities (see Milstein and Cuello, Nature 305: 537 (1983)), WO 93 / 08829 and Traunecker et al., EMBO J. 10: 3655 (1991)) and knob-in-hole manipulation (see, for example, U.S. Patent No. 5,731,168). Antibodies can also be made by manipulating electrostatic interaction effects to produce Fc antibody molecules (WO2009 / 089004A1); crosslinking of two or more antibodies or fragments (see, for example, U.S. Patent No. 4,676,980 and Brennan et al., Science 229: 81 (1985)); using leucine zippers to produce bispecific antibodies (see, for example, Kostelny et al., J. Immunol. 148(5): 1547-1553 (1992)); using diabody technology to make bispecific antibody fragments (see, for example, Hollinger et al., Proc.Natl. Acad. Sci. USA 90: 6444-6448 (1993)); and using single-chain Fv dimers (scFv) (see, for example, Gruber et al., J. Immunol. 152: 5368 (1994)); and preparing trispecific antibodies as described, for example, in Tutt et al., J. Immunol. 147: 60 (1991).

[00219] Manipulated antibodies with three or more functional antigen-binding sites, including Octopus antibodies, are also included here (see, for example, US document 2006 / 0025576).

[00220] The antibody or fragment thereof also includes a Dual Acting Fab or DAF comprising an antigen-binding site that binds to C5 as well as another different antigen (see document US2008 / 0069820, for example). 7. Variant Antibodies

[00221] In certain embodiments, variants of the amino acid sequence of the antibodies provided here are contemplated. By Petition 870260079322, dated 07 / 08 / 2026, p. 87 / 523 79 / 167 For example, it may be desirable to enhance the binding affinity and / or other biological properties of the antibody. Variants of the amino acid sequence of an antibody can be prepared by introducing appropriate modifications to the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and / or insertions and / or substitutions of residues within the antibody's amino acid sequences. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, for example, antigen binding. Variants with Replacement, Insertion and Deletion

[00222] In certain embodiments, antibody variants are provided that have one or more amino acid substitutions. Sites of interest for substitutional mutagenesis include the HVRs and FRs. Conservative substitutions are shown in Table 1 under the heading of preferred substitutions. More substantial alterations are provided in Table 1 under the heading of exemplary substitutions and as further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into an antibody of interest and the products screened for desired activity, e.g., maintained / enhanced antigen binding, decreased immunogenicity, or enhanced ADCC or CDC. Table 1 Original Residue Exemplary Substitutions Preferred Substitutions Wing (A) Val; Leu; Ile Val Arg (R) Lys; Gln; Asn Lys Asn (N) Gln; His; Asp; Lys; Arg Gln Asp (D) Glu; Asn Glu Cys (C) Ser; Only Ser Gln (Q) Asn; Glu Asn Glu (E) Asp; Gln Asp Petition 870260079322, of 07 / 08 / 2026, p. 88 / 523 80 / 167 Gly (G) Ala Ala His (H) Asn; Gln; Lys; Arg Arg Ile (I) Leu; Val; But; No; Phe; Norleucine Leu Leu (L) Norleucine; Ile; Val; But; No; Phe Ile Lys (K) Arg; Gln; Asn Arg Met (M) Leu; Phe; Ile Leu Phe (F) Trp; Leu; Val; Ile; No; Tyr Tyr Pro (P) Ala Ala Ser (S) Thr Thr Thr(T) Val; Ser Ser Trp (W) Tyr; Phe Tyr Tyr (Y) Trp; Phe; Thr; Ser Phe Val (V) Ile; Leu; But; Phe; No; Norleucine Leu

[00223] Amino acids can be grouped according to the common properties of the side chain: (1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[00224] Non-conservative substitutions will involve replacing a member of one of these classes with another class.

[00225] One type of substitution variant involves the substitution of one or more residues in the hypervariable region of a parental antibody (e.g., a human or humanized antibody). In general, the resulting variants selected for further study will have modifications (e.g., enhancements) in certain biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parental antibody and / or will substantially retain certain biological properties of the parental antibody. An exemplary substitution variant is an affinity-matured antibody which can be conveniently generated, for example, using phage-display-based affinity maturation techniques, such as those described herein. Briefly, one or more HVRs are mutated and the variant antibodies are displayed on phage and screened for biological activity in Petition 870260079322, dated 07 / 08 / 2026, page 89 / 523 81 / 167 particular (e.g., link affinity).

[00226] Alterations (e.g., substitutions) can be made to HVRs, for example, to improve antibody affinity. Such alterations can be made to HVR hotspots, i.e., codon-encoded residues that mutate at high frequency during the somatic maturation process (see, for example, Chowdhury, Methods Mol. Biol. 207: 179-196 (2008)) and / or antigen-contacting residues, with the resulting HV or VL variant being tested for binding affinity. Affinity maturation by constructing and re-selecting secondary libraries has been described, for example, in Hoogenboom et al., in Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)).In some affinity maturation modalities, diversity is introduced into the variable genes chosen for maturation through any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity. Another method for introducing diversity involves HVR-targeted approaches, in which multiple HVR residues (e.g., 4 to 6 residues at a time) are randomized. The HVR residues involved in antigen binding can be specifically identified, for example, using alanine-tracking mutagenesis or modeling. CDR-H3 and CDR-L3, in particular, are commonly targeted.

[00227] In certain embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, provided that such changes do not substantially reduce the antibody's ability to bind to the antigen. For example, conservative changes (e.g., conservative substitutions as provided) Petition 870260079322, dated 07 / 08 / 2026, p. 90 / 523 82 / 167 here) which do not substantially reduce binding affinity can be made to the HVRs. Such changes may, for example, be outside the antigen-contacting residues in the HVRs. In certain embodiments of the variant VH and VL sequences given above, each HVR is unaltered or contains no more than one, two, or three amino acid substitutions.

[00228] A useful method for identifying antibody residues or regions that can be targeted for mutagenesis is termed alanine-scavenging mutagenesis, as described by Cunningham, Science 244: 1081-1085 (1989). In this method, a target residue or group of residues (e.g., charged residues such as arg, asp, his, lys, and glu) are identified and replaced with neutral or negatively charged residues (e.g., alanine or polyalanine) to determine if the antibody-antigen interaction is affected. Additional substitutions can be introduced at amino acid locations, demonstrating functional sensitivity to the initial substitutions. Alternatively or additionally, a crystal structure of an antigen-antibody complex can identify contact points between the antibody and the antigen. Such contact residues and neighboring residues can be targeted or eliminated as candidates for substitution.Variants can be tracked to determine if they contain the desired properties.

[00229] Insertions into the amino acid sequence include fusions at the amino and / or carboxy terminus which vary in length from a single residue to polypeptides containing a hundred or more residues, as well as intra-sequence insertions of a single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue. Other variants of antibody molecule insertion include fusion of the N- or C-terminus of the antibody to an enzyme (e.g., for ADEPT) or a Petition 870260079322, dated 07 / 08 / 2026, p. 91 / 523 83 / 167 polypeptide that increases the serum half-life of the individual. b. Glycosylation Variants

[00230] In certain embodiments, an antibody is provided that is manipulated to increase or decrease the extent to which the antibody is glycosylated. The addition or elimination of glycosylation sites in an antibody can conveniently be achieved by altering the amino acid sequence so that one or more glycosylation sites are created or removed.

[00231] Where the antibody comprises an Fc region, the carbohydrate coupled to it can be altered. Native antibodies produced by mammalian cells typically comprise a biantenna branched oligosaccharide which is usually coupled by an N-linkage to Asn297 of the CH2 domain of the Fc region. See, for example, Wright et al., TIBTECH 15: 26-32 (1997). The oligosaccharide may include various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as a fucose coupled to a GlcNAc in the trunk of the biantenna oligosaccharide structure. In some embodiments, modifications of the oligosaccharide in an antibody of the invention can be made in order to create antibody variants with certain enhanced properties.

[00232] In one embodiment, the antibody variants that are provided have a carbohydrate structure that does not exhibit fucose coupled (directly or indirectly) to an Fc region. For example, the amount of fucose in such an antibody may be between 1% and 80%, between 1% and 65%, between 5% and 65%, or between 20% and 40%. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain in Asn297 relative to the sum of all glyco-structures coupled to Asn297 (e.g., complex, hybrid, and mannose-rich structures), as measured by MALDI-TOF mass spectrometry, as described in the document. Petition 870260079322, dated 07 / 08 / 2026, page 92 / 523 84 / 167 mento WO2008 / 077546, for example. Asn297 refers to the asparagine residue located around position 297 in the Fc region (I numbering of residues in the Fc region); however, Asn297 may also be located approximately + / - 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to small sequence variations in the antibodies. Such fucosylation variants may have increased ADCC function. See, for example, U.S. Patent Publications Nos. US 2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd.). Examples of publications related to defucosylated or fucose-deficient antibody variants include: documents US 2003 / 0157108; WO2000 / 61739; WO2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US 2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO2003 / 085119; WO2003 / 084570; WO2005 / 035586; WO2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., J. Mol. Biol.336: 1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng. 87:614 (2004). Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells deficient in fucosylation protein (Ripka et al., Arch. Biochem. Biophys. 249: 533-545 (1986); US document 2003 / 0157108, Presta, L; and WO2004 / 056312, Adams et al., especially in Example 11) and knockout cell lines, such as CHO cells silenced for the alpha-1,6-fucosyltransferase gene, FUT8 (see, for example, Yamane-Ohnuki et al., Biotech. Bioeng. 87: 614 (2004); Kanda et al., Biotechnol. Bioeng. 94(4): 680-688 (2006); and WO2003 / 085107).

[00233] Additionally, antibody variants with bifurcated oligosaccharides are provided, for example, in which a biantennary oligosaccharide coupled to the Fc region of the antibody is bifurcated by GlcNAc. Such antibody variants may have reduced fucosylation. Petition 870260079322, dated 07 / 08 / 2026, p. 93 / 523 85 / 167 and / or enhanced ADCC function. Examples of such antibody variants are described, for example, in document WO2003 / 011878 (Jean Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and document US 2005 / 0123546 (Umana et al.). Antibody variants with at least one galactose residue in the oligosaccharide coupled to the Fc region are also provided. Such antibody variants may have enhanced CDC function. Such antibody variants are described, for example, in document WO1997 / 30087 (Patel et al.); document WO1998 / 58964 (Raju, S.); and document WO1999 / 22764 (Raju, S.). c. Region Variants Fc

[00234] In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3, or IgG4 region) comprising an amino acid modification (e.g., a substitution) at one or more amino acid positions.

[00235] In certain embodiments, the invention considers an antibody variant that has some, but not all, effector functions, making it a desirable candidate for applications where the antibody's in vivo half-life is important, while certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be conducted to confirm the reduction / or depletion of CDC and / or ADCC activities. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks binding to Fc gamma R9 (thus likely lacking ADCC activity) but retains the ability to bind to FcRn. Primary cells for mediation of Petition 870260079322, dated 07 / 08 / 2026, p. 94 / 523 86 / 167 ADCC, NK cells, express only Fc gamma RIII, while monocytes express Fc gamma RI, Fc gamma RII, and Fc gamma RIII. FcR expression in hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9: 457-492 (1991). Non-limiting examples of in vitro assays to evaluate the ADCC activity of a molecule of interest are described in U.S. Patent No. 5,500,362 (see, for example, Hellstrom et al., Proc. Nat'l Acad. Sci. USA 83: 7059-7063 (1986)) and Hellstrom et al., Proc. Nat'l Acad. Sci. USA 82: 1499-1502 (1985); United States Patent No. 5,821,337 (see Bruggemann et al., J. Exp. Med. 166: 1351-1361 (1987)). Alternatively, non-radioactive assay methods may be employed (see, for example, ACTI™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc., Mountain View, CA); and CytoTox 96 non-radioactive cytotoxicity assay (trademark) (Promega, Madison, WI)).Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of the molecule of interest can be evaluated in vivo, for example, in an animal model, such as that described in Clynes et al., Proc. Nat'l Acad. Sci. USA 95: 652-656 (1998). C1q binding assays can also be performed to confirm that the antibody is unable to bind to C1q and therefore has no CDC activity. See, for example, C1q and C3c binding ELISA in WO2006 / 029879 and WO2005 / 100402. To assess complement activation, a CDC assay can be performed (see, for example, GazzanoSantoro et al., J. Immunol. Methods 202: 163 (1996); Cragg et al., Blood 101: 1045-1052 (2003); and Cragg et al., Blood 103: 2738-2743 (2004)).Binding to FcRn and in vivo determinations of elimination / half-life can also be performed using co-op methods. Petition 870260079322, dated 07 / 08 / 2026, p. 95 / 523 87 / 167 known in the art (see, for example, Petkova et al., Int'l. Immunol. 18(12): 1759-1769 (2006)).

[00236] Antibodies with reduced effector function include those with substitution of one or more of the residues in the Fc region 238, 265, 269, 270, 297, 327, and 329 (U.S. Patent No. 6,737,056). Such Fc mutants include mutants with substitutions at two or more amino acid positions 265, 269, 270, 297, and 327, including the so-called Fc mutant DANA, with substitution of residues 265 and 297 for alanine (U.S. Patent No. 7,332,581).

[00237] Certain antibody variants with increased or decreased binding to FcRs are described. (See, for example, U.S. Patent No. 6,737,056; document WO2004 / 056312 and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)).

[00238] In certain embodiments, an antibody variant comprises an Fc region with one or more amino acid substitutions that increase ADCC, for example, substitutions at positions 298, 333 and / or 334 of the Fc region (EU numbering of residues).

[00239] In some embodiments, changes are made to the Fc region that result in C1q binding (i.e., increased or decreased) and / or altered Complement-Dependent Cytotoxicity (CDC), for example, as described in U.S. Patent No. 6,194,551, document WO1999 / 51642 and Idusogie et al., J. Immunol. 164: 4178-4184 (2000).

[00240] Antibodies with increased half-lives and enhanced binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117: 587 (1976) and Kim et al., J. Immunol. 24: 249 (1994)), are described in document US2005 / 0014934 (Hinton et al.). These antibodies comprise an Fc region with one or more substitutions that enhance Petition 870260079322, dated 07 / 08 / 2026, p. 96 / 523 88 / 167 linking the Fc region to FcRn. Such Fc variants include those with substitutions in one or more residues of the Fc region: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424 or 434, for example, substitution of residue 434 of the Fc region (United States Patent No. 7,371,826).

[00241] See also Duncan, Nature 322: 738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821; and document WO1994 / 29351 regarding other examples of Fc region variants.

[00242] d. Cysteine-Manipulated Antibody Variants

[00243] In certain embodiments, it may be desirable to produce cysteine-manipulated antibodies, for example, tioMAbs, in which one or more residues of an antibody are replaced by cysteine ​​residues. In particular embodiments, the substituted residues occur at accessible sites of the antibody. By replacing these residues with cysteine, reactive thiol groups are thus positioned at accessible sites of the antibody and can be used to conjugate the antibody to other moieties, such as drug moieties or drug-ligand moieties, to create an immunoconjugate, as described later herein. In certain embodiments, any one or more of the following residues may be replaced by cysteine: C205 (Kabat numbering) of the light chain; A118 (EU numbering) of the heavy chain and S400 (EU numbering) of the Fc region of the heavy chain.Cysteine-based antibodies can be generated as described, for example, in U.S. Patent No. 7,521,541. e. Antibody Derivatives

[00244] In certain embodiments, an antibody provided herein may be further modified to contain additional non-proteinaceous moieties which are known in the art and readily available. The moieties suitable for antibody derivatization Petition 870260079322, dated 07 / 08 / 2026, page 97 / 523 89 / 167 includes, but is not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (such as homopolymers or random copolymers) and dextran or poly(n-vinylpyrrolidone)polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyethoxylated polyols (e.g., glycerol), polyvinyl alcohol and mixtures thereof. Polyethylene glycol propionaldehyde may have many advantages in manufacturing due to its stability in water. The polymer can be of any molecular weight and can be branched or unbranched.The number of polymers coupled to the antibody can vary, and if more than one polymer is coupled, they may be the same or different molecules. In general, the number and / or type of polymers used for derivatization can be determined based on considerations that include, but are not limited to, the particular properties or functions of the antibody to be enhanced, whether the antibody derivative will be used in therapy under defined conditions, etc.

[00245] In another embodiment, antibody conjugates and a non-proteinaceous moiety are provided that can be selectively heated by radiation exposure. In one embodiment, the non-proteinaceous moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation can be of any wavelength and includes, but is not limited to, wavelengths that do not damage ordinary cells but heat the non-proteinaceous moiety to a temperature at which cells near the antibody-non-proteinaceous moiety are killed. B. Recombinant Methods and Compositions Petition 870260079322, dated 07 / 08 / 2026, p. 98 / 523 90 / 167

[00246] Antibodies can be produced using recombinant methods and compositions, for example, as described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding an anti-C5 antibody described herein is provided. Such nucleic acid may encode an amino acid sequence comprising VL and / or an amino acid sequence comprising VH of the antibody (e.g., the light and / or heavy chains of the antibody). In a further embodiment, one or more vectors (e.g., expression vectors) comprising such nucleic acid are provided.In one such embodiment, a host cell comprises (e.g., has been transformed with): (1) a vector comprising an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antibody or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and a second vector comprising a nucleic acid encoding an amino acid sequence comprising the VH of the antibody. In one embodiment, the host cell is eukaryotic, e.g., Chinese Hamster Ovary (CHO) cell or a lymphoid cell (e.g., Y0, NS0, Sp20 cell).In one embodiment, a method is provided for the production of an anti-C5 antibody, wherein the method comprises culturing a host cell comprising a nucleic acid encoding an antibody as provided above under conditions suitable for antibody expression and, optionally, recovering the antibody from the host cell (or from the host cell culture medium).

[00247] For the recombinant production of an anti-C5 antibody, a nucleic acid encoding an antibody, for example, as described above, is isolated and inserted into one or more vectors for cloning and / or further expression in a host cell. Such nucleic acid Petition 870260079322, dated 07 / 08 / 2026, page 99 / 523 91 / 167 co can be readily isolated and sequenced using conventional procedures (e.g., using oligonucleotide probes that are able to specifically bind to the genes encoding the heavy and light chains of the antibody).

[00248] Suitable host cells for cloning and expression of antibody-encoding vectors include the prokaryotic or eukaryotic cells described herein. For example, antibodies can be produced in bacteria, particularly when glycosylation and Fc effector function are not required. For expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Patents Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pages 245–254, which describes the expression of antibody fragments in E. coli). After expression, the antibody can be isolated from the bacterial cell suspension in a soluble fraction and can be further purified.

[00249] In addition to prokaryotes, eukaryotic microbes, such as filamentous fungi or yeast, are suitable hosts for cloning and expression of antibody-encoding vectors, including fungal and yeast strains whose glycosylation pathways have been humanized, resulting in the production of an antibody with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004) and Li et al., Nat. Biotech. 24: 210-215 (2006).

[00250] Host cells suitable for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified that can be used in conjunction with cells. Petition 870260079322, dated 07 / 08 / 2026, page 100 / 523 92 / 167 insect squid, particularly for the transformation of Spodoptera frugiperda cells.

[00251] Plant cell cultures can also be used as hosts. See, for example, U.S. Patents Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978 and 6,417,429 (which describe PLANTIBODIES™ technology for producing antibodies in transgenic plants).

[00252] Vertebrate cells can also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension can be useful. Other examples of mammalian host cell lines are SV40-transformed monkey kidney CV1 cell line (COS-7); human embryonic kidney cell line (293 or 293 cells as described, for example, in Graham et al., J. Gen Virol. 36: 59 (1977)); hamster pup kidney cells (BHK); mouse Sertoli cells (TM4 cells as described, for example, in Mather, Biol. Reprod. 23: 243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); dog kidney cells (MDCK); buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells as described, for example, in Mather et al., Annals NY Acad. Sci.383: 44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian cell lines include Chinese hamster ovary (CHO) cells, including CHO DHFR cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77: 4216 (1980)); and myeloma cell lines such as Y0, NS0, and Sp2 / 0. For a review of other useful mammalian cell lines for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ), pp. 255-268. Petition 870260079322, dated 07 / 08 / 2026, page 101 / 523 93 / 167 (2003).

[00253] Polyclonal antibodies are preferably generated in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, for example, keyhole lympet hemocyanin, serum albumin, bovine thyroglobulin or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, sulfosuccinimide maleimidobenzoyl ester (conjugation via cysteine ​​residues), N-hydroxysuccinimide (via lysine residues), glutaraldehyde, succinic anhydride, SOCl2 or R1N=C=NR, where R and R1 are different alkyl groups.

[00254] Animals (generally non-human mammals) are immunized against the antigen, immunogenic conjugates, or derivatives by combining, for example, 100 micrograms or 5 micrograms of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites. One month later, the animals are inoculated again with 1 / 5 to 1 / 10 of the original amount at multiple sites. Seven to 14 days later, the animals are bled and the serum is assayed for antibody titer. Animals are inoculated until the titer reaches a plateau. Preferably, the animal is inoculated with the conjugate of the same antigen, but conjugated to a different protein and / or through a different crosslinking reagent. Conjugates can also be made in recombinant cell culture as fusion proteins.Aggregating agents, such as alum, are suitable for use in enhancing the immune response.

[00255] Monoclonal antibodies are obtained from a substantially homogeneous population of antibodies, that is, the antibodies Petition 870260079322, dated 07 / 08 / 2026, page 102 / 523 94 / 167 individual posocytes comprising the population are identical except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerizations, amidations) that may be present in small amounts. Therefore, the monoclonal modifier indicates the antibody's character as not being a mixture of distinct antibodies.

[00256] For example, monoclonal antibodies can be made using the hybridoma method first described by Kohler et al. Nature 256(5517): 495-497 (1975). In the hybridoma method, a mouse or other suitable host animal, such as a hamster, is immunized as described above to generate lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes can be immunized in vitro.

[00257] The immunization agent will typically include the antigenic protein or a fusion variant thereof. Generally, peripheral blood lymphocytes (PBLs) are used if cells of human origin are desired, or spleen or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusion agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice, Academic Press (1986), pp. 59-103).

[00258] Immortalized cell lines are generally transformed mammalian cells, particularly rodent, bovine, and human myeloma cells. Rat or mouse myeloma cell lines are usually employed. The hybridoma cells thus prepared are seeded and cultured in a suitable culture medium that preferably contains one or more substances that inhibit cell growth or survival. Petition 870260079322, dated 07 / 08 / 2026, page 103 / 523 95 / 167 of parental myeloma cells, unfused. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT), the culture medium for hybridomas will typically include hypoxanthine, aminopterin, and thymidine (HAT medium), which are substances that prevent the growth of HGPRT-deficient cells.

[00259] Preferred immortalized myeloma cells are those that fuse efficiently, support stable high-level antibody production by selected antibody-producing cells, and are sensitive to media such as HAT medium. Among these, murine myeloma cell lines are preferred, such as those derived from MOPC-21 and MPC-11 mouse tumors, made available by the Salk Institute Cell Distribution Center, San Diego, California, USA, and SP-2 cells (and their derivatives, e.g., X63-Ag8-653) made available by the American Type Culture Collection, Manassas, Virginia, USA. Human myeloma and mouse-human heteromyeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor et al., J Immunol. 133(6): 3001-3005 (1984); Brodeur et al., Techniques and Applications of Monoclonal Antibody Production, Marcel Dekker, Inc., New York (1987), pages 51-63).

[00260] The culture medium in which the hybridoma cells are grown is assayed for the production of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of the monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as a radioimmunoassay (RIA) or an enzyme-linked immunosorbent assay (ELISA). Such techniques and assays are known. For example, binding affinity can be determined by the Scatchard analysis of Munson, Anal. Biochem. 107(1): 220-239 (1980). Petition 870260079322, dated 07 / 08 / 2026, page 104 / 523 96 / 167

[00261] After antibody-producing hybridoma cells with the desired specificity, affinity, and / or activity are identified, clones can be subcloned using limiting dilution procedures and cultured using standardized methods (Goding, supra). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 media. Additionally, hybridoma cells can be cultured in vivo as tumors in a mammal.

[00262] Monoclonal antibodies secreted by subclones are adequately separated from culture medium, ascites fluid or serum by conventional immunoglobulin purification procedures such as, for example, chromatography with Sepharose A protein, hydroxyapatite, gel electrophoresis, dialysis or affinity chromatography.

[00263] Antibodies can be produced by immunizing a suitable host animal against an antigen. In one embodiment, the antigen is a polypeptide comprising a full-length C5. In one embodiment, the antigen is a polypeptide comprising the beta chain (: SEQ ID NO: 40) of C5. In one embodiment, the antigen is a polypeptide comprising the MG1-MG2 domain (: SEQ ID NO: 43) of the beta chain of C5. In one embodiment, the antigen is a polypeptide comprising the MG1 domain (: SEQ ID NO: 41) of the beta chain of C5. In one embodiment, the antigen is a polypeptide comprising the region corresponding to amino acids at positions 19 to 180 of the beta chain of C5. In one embodiment, the antigen is a polypeptide comprising the region corresponding to amino acids at positions 33 to 124 of the beta chain of C5.In one embodiment, the antigen is a polypeptide comprising at least one fragment selected from amino acids 47-57, 70-76, and 107-110 of the beta chain (: SEQ ID NO: 40). Petition 870260079322, dated 07 / 08 / 2026, page 105 / 523 97 / 167 of C5. In one embodiment, the antigen is a polypeptide comprising a C5 beta chain fragment comprising at least one amino acid selected from the group consisting of Thr47, Glu48, Ala49, Phe50, Asp51, Ala52, Thr53, Lys57, His70, Val71, His72, Ser74, Glu76, Val107, Ser108, ​​Lys109, and His110. In another embodiment, the antigen is a polypeptide comprising a C5 beta chain fragment comprising at least one amino acid selected from the group consisting of Glu48, Asp51, His70, His72, Lys109, and His110. Also included in the present invention are antibodies produced by immunizing an animal against the antigen. The antibodies may incorporate any of the features, individually or in combination, as described in Exemplary Anti-C5 Antibodies above. C. Essays

[00264] The anti-C5 antibodies provided here can be identified, screened, or characterized for their physical / chemical properties and / or biological activities by various assays known in the art. 1. Bonding Tests and Other Tests

[00265] In one aspect, an antibody of the invention is tested for its antigen-binding activity, for example, by means of known methods such as ELISA, Western blot, BIACORE (registered trademark), etc.

[00266] In another aspect, competition assays can be used to identify an antibody that competes for binding to C5 with an anti-C5 antibody described herein. In certain embodiments, when such a competing antibody is present in excess, it blocks (i.e., reduces) binding to a reference antibody for C5 by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or more. In some circumstances Petition 870260079322, dated 07 / 08 / 2026, p. 106 / 523 In 98 / 167 instances, binding is inhibited by at least 80%, 85%, 90%, 95% or more. In certain embodiments, such competing antibodies bind to the same epitope (e.g., a linear or conformational epitope) that is bound by the anti-C5 antibody described herein (e.g., an anti-C5 antibody described in Table 2). Detailed exemplary methods for mapping an epitope to which an antibody binds are provided in Morris, Epitope Mapping Protocols, in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ) (1996).

[00267] In an exemplary competition assay, immobilized C5 is incubated in a solution comprising a first labeled (reference) antibody that binds to C5 and a second unlabeled antibody that is being tested for its ability to compete with the first antibody for binding to C5. The second antibody may be present in a hybridoma supernatant. As a control, immobilized C5 is incubated in a solution comprising the first labeled antibody but not the second unlabeled antibody. After incubation under conditions permissive for binding of the first antibody to C5, the excess unbound antibody is removed and the amount of marker associated with immobilized C5 is measured. If the amount of marker associated with immobilized C5 is substantially reduced in the test sample compared to the control sample, this indicates that the second antibody is competing with the first antibody for binding to C5.See Harlow and Lane, Antibodies: A Laboratory Manual, Chapter 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY) (1988).

[00268] In another exemplary competition assay, BIACORE (registered trademark) is used to determine the ability of a test anti-C5 antibody to compete for binding to C5 with a second (reference) anti-C5 antibody. In a further aspect, in which Petition 870260079322, dated 07 / 08 / 2026, p. 107 / 523 99 / 167 A BIACORE (registered trademark) instrument (e.g., BIACORE (registered trademark) 3000) is operated according to the manufacturer's recommendations. The C5 protein is captured on a BIACORE (registered trademark) CM5 chip using a known standardized technique to generate a C5-coated surface. Typically, 200-800 C5 resonance units will be coupled to the chip (an amount that gives easily measurable levels of binding, but which are rapidly saturated by the concentrations of the assay antibody to be used). The two antibodies (i.e., the assay antibody and the reference antibody) to be evaluated for their ability to compete with each other are mixed in a 1:1 molar ratio of binding sites in a suitable buffer to create a test mixture.When concentrations based on binding sites are calculated, the molecular weight of an assay or reference antibody is assumed to be the total molecular weight of the corresponding antibody divided by the number of C5 binding sites on the antibody. The concentration of each antibody (i.e., the assay and reference antibodies) in the test mixture must be high enough to rapidly saturate the binding sites for that antibody on the C5 molecules captured on the BIACORE chip (trademark). The assay and reference antibodies in the mixture are at the same molar concentration (based on binding), typically between 1.00 and 1.5 micromolar (based on binding site). Separate solutions containing only the assay antibody and only the reference antibody are also prepared. The assay antibody and the reference antibody in these solutions must be in the same buffer and at the same concentration and under the same conditions as in the test mixture.The test mixture containing the assay antibody and the reference antibody is passed over the C5-coated BIACORE chip (trademark) and the total amount of binding is recorded. The chip is then treated. Petition 870260079322, dated 07 / 08 / 2026, page 108 / 523 100 / 167 method to remove the bound assay antibody or reference antibody without damaging the C5-coated chip. Typically, this is done by treating the chip with 30 mM HCl for 60 seconds. The assay antibody solution alone is then passed over the C5-coated surface and the amount of binding is recorded. The chip is treated again to remove all bound antibody without damaging the C5-coated chip. The reference antibody solution alone is then passed over the C5-coated surface and the amount of binding is recorded. The chip is treated again to remove all bound antibody without damaging the C5-coated chip. The maximum theoretical binding of the assay antibody and reference antibody mixture is then calculated and is the sum of the binding of each antibody (i.e., assay and reference) when passed only over the C5 surface.If the actual recorded binding of the mixture is less than this theoretical maximum, then the assay antibody and the reference antibody are competing with each other for binding to C5. Therefore, in general, a competing anti-C5 antibody is one that will bind to C5 in the above blocking assay (trademark) such that during the assay and in the presence of the reference anti-C5 antibody, the recorded binding is between 80% and 0.1% (e.g., 80% > 4%) of the theoretical maximum binding, specifically between 75% and 0.1% (e.g., 75% to 4%) of the theoretical maximum binding, and more specifically between 70% and 0.1% (e.g., 70% to 4%) of the theoretical maximum binding (as defined above) of the assay antibody and the reference antibody in the combination.

[00269] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from antibodies CFA0341 and CFA0330. In some embodiments, an anti-C5 antibody competes for binding to C5 with an antibody selected from: Petition 870260079322, dated 07 / 08 / 2026, p. 109 / 523 101 / 167 CFA0538, CFA0501, CFA0599, CFA0307, ​​CFA0366, CFA0675, and CFA0672. In some embodiments, an anti-C5 antibody competes for binding to C5 with antibody CFA0329. In some embodiments, an anti-C5 antibody competes for binding to C5 with antibody CFA0666.

[00270] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair of the CFA0305 or 305LO5 antibody.

[00271] In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a neutral pH than at an acidic pH. In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from: CFA0538, CFA0501, CFA0599, CFA0307, ​​CFA0366, CFA0675, and CFA0672. In some embodiments, an anti-C5 antibody competes for binding to C5 with the CFA0666 antibody. In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[00272] In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a neutral pH than at an acidic pH. In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair of the CFA0305 or 305LO5 antibody. In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[00273] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from a VH of SEQ ID: SEQ ID NO: 22 and a VL of SEQ ID: SEQ ID NO: 26 or a VH of SEQ ID: SEQ ID NO: 21 and a VL of SEQ ID: SEQ ID Petition 870260079322, dated 07 / 08 / 2026, p. 110 / 523 102 / 167 NO: 25. In some embodiments, an anti-C5 antibody competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID : SEQ ID NO: 5 and a VL of SEQ ID : SEQ ID NO: 15; (b) a VH of SEQ ID : SEQ ID NO: 4 and a VL of SEQ ID : SEQ ID NO: 14; (c) a VH of SEQ ID : SEQ ID NO: 6 and a VL of SEQ ID : SEQ ID NO: 16; (d) a VH of SEQ ID : SEQ ID NO: 2 and a VL of SEQ ID : SEQ ID NO: 12; (e) a VH of SEQ ID : SEQ ID NO: 3 and a VL of SEQ ID : SEQ ID NO: 13; (f) a VH of SEQ ID : SEQ ID NO: 1 and a VL of SEQ ID : SEQ ID NO: 11; (g) a VH of SEQ ID : SEQ ID NO: 9 and a VL of SEQ ID : SEQ ID NO: 19; (h) a VH of SEQ ID : SEQ ID NO: 7 and a VL of SEQ ID : SEQ ID NO: 17; and (i) a VH of SEQ ID : SEQ ID NO: 8 and a VL of SEQ ID : SEQ ID NO: 18. In some embodiments, an anti-C5 antibody competes for binding to C5 with an antibody comprising a VH of SEQ ID : SEQ ID NO: 23 and a VL of SEQ ID : SEQ ID NO: 27.In some embodiments, an anti-C5 antibody competes for binding to C5 with an antibody comprising a VH of SEQ ID: SEQ ID NO: 7 and a VL of SEQ ID: SEQ ID NO: 17.

[00274] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID : SEQ ID NO: 1 and a VL of SEQ ID : SEQ ID NO: 11; (b) a VH of SEQ ID : SEQ ID NO: 22 and a VL of SEQ ID : SEQ ID NO: 26; (c) a VH of SEQ ID : SEQ ID NO: 21 and a VL of SEQ ID : SEQ ID NO: 25; (d) a VH of SEQ ID : SEQ ID NO: 5 and a VL of SEQ ID : SEQ ID NO: 15; (e) a VH of SEQ ID : SEQ ID NO: 4 and a VL of SEQ ID : SEQ ID NO: 14; (f) a VH of SEQ ID: SEQ ID NO: 6 and a VL of SEQ ID: SEQ ID NO: 16; (g) a VH of SEQ ID: SEQ ID NO: 2 and a VL of SEQ ID: SEQ ID NO: 12; (h) a VH of Petition 870260079322, dated 07 / 08 / 2026, page 111 / 523 103 / 167 SEQ ID : SEQ ID NO: 3 e uma VL de SEQ ID : SEQ ID NO: 13; (i) uma VH de SEQ ID : SEQ ID NO: 9 e uma VL de SEQ ID : SEQ ID NO: 19; (j) uma VH de SEQ ID : SEQ ID NO: 7 e uma VL de SEQ ID : SEQ ID NO: 17; (k) uma VH de SEQ ID : SEQ ID NO: 8 e uma VL de SEQ ID : SEQ ID NO: 18; (l) uma VH de SEQ ID : SEQ ID NO: 23 e uma VL de SEQ ID : SEQ ID NO: 27; e (m) uma VH de SEQ ID : SEQ ID NO: 10 e uma VL de SEQ ID : SEQ ID NO: 20.

[00275] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID : SEQ ID NO: 22 and a VL of SEQ ID : SEQ ID NO: 26; (b) a VH of SEQ ID : SEQ ID NO: 21 and a VL of SEQ ID : SEQ ID NO: 25; (c) a VH of SEQ ID : SEQ ID NO: 5 and a VL of SEQ ID : SEQ ID NO: 15; (d) a VH of SEQ ID : SEQ ID NO: 4 and a VL of SEQ ID : SEQ ID NO: 14; (e) a VH of SEQ ID : SEQ ID NO: 6 and a VL of SEQ ID : SEQ ID NO: 16; (f) a VH of SEQ ID: SEQ ID NO: 2 and a VL of SEQ ID: SEQ ID NO: 12; (g) a VH of SEQ ID: SEQ ID NO: 3 and a VL of SEQ ID: SEQ ID NO: 13; (h) a VH of SEQ ID: SEQ ID NO: 9 and a VL of SEQ ID: SEQ ID NO: 19; (i) a VH of SEQ ID: SEQ ID NO: 7 and a VL of SEQ ID: SEQ ID NO: 17; (j) a VH of SEQ ID: SEQ ID NO: 8 and a VL of SEQ ID: SEQ ID NO: 18; (k) a VH of SEQ ID: SEQ ID NO: 23 and a VL of SEQ ID: SEQ ID NO: 27.

[00276] In certain embodiments, an anti-C5 antibody of the present invention competes for binding to C5 with an antibody comprising a VH and VL pair selected from a VH of SEQ ID : SEQ ID NO: 1 and a VL of SEQ ID : SEQ ID NO: 11 or a VH of SEQ ID : SEQ ID NO: 10 and a VL of SEQ ID : SEQ ID NO: 20.

[00277] In additional embodiments, the anti-C5 antibody binds to Petition 870260079322, dated 07 / 08 / 2026, p. 112 / 523 104 / 167 C5 has a greater affinity for neutral pH than for acidic pH.In certain embodiments, an anti-C5 antibody binds to C5 with a higher affinity at neutral than at acidic pH and competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID : SEQ ID NO: 1 and a VL of SEQ ID : SEQ ID NO: 11; (b) a VH of SEQ ID : SEQ ID NO: 5 and a VL of SEQ ID : SEQ ID NO: 15; (c) a VH of SEQ ID : SEQ ID NO: 4 and a VL of SEQ ID : SEQ ID NO: 14; (d) a VH of SEQ ID : SEQ ID NO: 6 and a VL of SEQ ID : SEQ ID NO: 16; (e) a VH of SEQ ID : SEQ ID NO: 2 and a VL of SEQ ID : SEQ ID NO: 12; (f) a VH of SEQ ID: SEQ ID NO: 3 and a VL of SEQ ID: SEQ ID NO: 13; (g) a VH of SEQ ID: SEQ ID NO: 9 and a VL of SEQ ID: SEQ ID NO: 19; (h) a VH of SEQ ID: SEQ ID NO: 7 and a VL of SEQ ID: SEQ ID NO: 17; (i) a VH of SEQ ID: SEQ ID NO: 8 and a VL of SEQ ID: SEQ ID NO: 18; and (j) a VH of SEQ ID: SEQ ID NO: 10 and a VL of SEQ ID: SEQ ID NO: 20.In additional embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[00278] In some embodiments, the anti-C5 antibody binds to C5 with a higher affinity at neutral than at acidic pH and competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID : SEQ ID NO: 5 and a VL of SEQ ID : SEQ ID NO: 15; (b) a VH of SEQ ID : SEQ ID NO: 4 and a VL of SEQ ID : SEQ ID NO: 14; (c) a VH of SEQ ID : SEQ ID NO: 6 and a VL of SEQ ID : SEQ ID NO: 16; (d) a VH of SEQ ID : SEQ ID NO: 2 and a VL of SEQ ID : SEQ ID NO: 12; (e) a VH of SEQ ID: SEQ ID NO: 3 and a VL of SEQ ID: SEQ ID NO: 13; (f) a VH of SEQ ID: SEQ ID NO: 1 and a VL of SEQ ID: SEQ ID NO: 11; (g) a VH of SEQ ID: SEQ ID NO: 9 and a VL of SEQ ID: SEQ ID NO: 19; (h) a VH of SEQ ID: SEQ ID NO: 7 and Petition 870260079322, dated 07 / 08 / 2026, page 113 / 523 105 / 167 a VL of SEQ ID : SEQ ID NO: 17; and (i) a VH of SEQ ID : SEQ ID NO: 8 and a VL of SEQ ID : SEQ ID NO: 18. In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[00279] In some embodiments, the anti-C5 antibody binds to C5 with a higher affinity at neutral than at an acidic pH and competes for binding to C5 with an antibody comprising a VH and VL pair selected from a VH of SEQ ID : SEQ ID NO: 1 and a VL of SEQ ID : SEQ ID NO: 11 or a VH of SEQ ID : SEQ ID NO: 10 and a VL of SEQ ID : SEQ ID NO: 20. In further embodiments, the anti-C5 antibody binds to C5 with a higher affinity at a pH of 7.4 than at a pH of 5.8.

[00280] In certain embodiments, it can be determined whether an anti-C5 antibody of the present invention binds to a particular epitope as follows: C5 point mutants in which an amino acid (except alanine) of C5 is replaced by alanine are expressed in cells 293 and the binding of an anti-C5 antibody to the C5 mutants is tested by ELISA, Western blot or BIACORE (trademark); wherein a substantial reduction or elimination of binding of the anti-C5 antibody to the C5 mutant relative to its binding to wild-type C5 indicates that the anti-C5 antibody binds to an epitope comprising that amino acid on C5. In certain embodiments, the amino acid on C5 to be replaced by alanine is selected from the group consisting of Glu48, Asp51, His70, His72, Lys109 and His110 of the beta chain of C5 (: SEQ ID NO: 40). In additional embodiments, the amino acid on C5 to be replaced by alanine is Asp51 or Lys109 of the beta chain of C5 (: SEQ ID NO: 40).

[00281] In certain embodiments, it can be determined whether an anti-C5 antibody with pH-dependent binding characteristics binds to a given epitope as follows: point mutants Petition 870260079322, dated 07 / 08 / 2026, p. 114 / 523 106 / 167 C5 mutants in which a histidine residue is replaced by another amino acid (e.g., tyrosine) are expressed in cells 293 and the binding of an anti-C5 antibody to the C5 mutants is tested by ELISA, Western blot, or BIACORE (trademark); wherein a substantial reduction in the binding of the anti-C5 antibody to wild-type C5 at an acidic pH relative to its binding to the C5 mutant at an acidic pH indicates that the anti-C5 antibody binds to an epitope comprising that histidine residue on C5. In further embodiments, the binding of the anti-C5 antibody to wild-type C5 at a neutral pH is not substantially reduced relative to its binding to the C5 mutant at a neutral pH. In certain embodiments, the histidine residue on C5 to be replaced by another amino acid is selected from the group consisting of His70, His72, and His110 of the beta chain of C5 (: SEQ ID NO: 40).In an additional embodiment, the histidine residue His70 is replaced by tyrosine. 2. Activity Trials

[00282] In one aspect, assays are provided to identify anti-C5 antibodies that have biological activity. Biological activity may include, for example, inhibition of C5 activation, prevention of C5 cleavage to form C5a and C5b, blocking of C5 convertase access to the cleavage site on C5, blocking of hemolytic activity caused by C5 activation, etc. Antibodies with such biological activity in vivo and / or in vitro are also provided.

[00283] In certain embodiments, an antibody of the invention is tested for such biological activity.

[00284] In certain embodiments, it is determined whether a test antibody inhibits the cleavage of C5 into C5a and C5b by means of the methods described, for example, in Isenman et al., J Immunol. 124(1): 326-331 (1980). In another embodiment, this is determined by means of Petition 870260079322, dated 07 / 08 / 2026, p. 115 / 523 107 / 167 methods for specific detection of cleaved C5a and / or C5b proteins, for example, ELISAs or Western blots. Where a decreased amount of a C5 cleavage product (i.e., C5a and / or C5b) is detected in the presence of (or after contact with) the test antibody, the test antibody is detected as an antibody that can inhibit C5 cleavage. In certain embodiments, the concentration and / or physiological activity of C5a can be measured by methods, for example, chemotaxis assays, RIAs or ELISAs (see, for example, Ward and Zvaifler J. Clin. Invest. 50(3): 606-616 (1971)).

[00285] In certain embodiments, it is determined whether a test antibody blocks C5 convertase access to C5 by means of methods for detecting protein interactions between C5 convertase and C5, for example, ELISAs or BIACORES (registered trademark). Where the interactions are diminished in the presence (or after contact with) the test antibody, the test antibody is identified as an antibody that can block C5 convertase access to C5.

[00286] In certain embodiments, C5 activity can be measured as a function of its ability to subject cells to lysis in an individual's body fluids. The ability to lyse cells or its reduction by C5 can be measured by methods well known in the art, for example, a conventional hemolytic assay, such as a hemolysis assay described by Kabat and Mayer (eds.), Experimental Immunochemistry, 2nd Edition, 135-240, Springfield, IL, CC Thomas (1961), pages 135-139 or a conventional variation of this assay, such as the chicken erythrocyte hemolysis method described, for example, in Hillmen et al., N. Engl. J. Med. 350(6): 552-559 (2004). In certain embodiments, C5 activity or its inhibition is quantified using a CH50eq assay. The CH50eq assay is a method for measuring total complement activity in serum. This test is a lytic assay that uses erythrocytes. Petition 870260079322, dated 07 / 08 / 2026, page 116 / 523 108 / 167 sensitized by the antibody as the activator of the classical complement pathway and various dilutions of the test serum to determine the amount required to allow 50% lysis (CH50). The percentage of hemolysis can be determined, for example, using a spectrophotometer. The CH50eq assay provides an indirect measure of terminal complement complex (TCC) formation, since TCC itself is directly responsible for the measured hemolysis. Inhibition of C5 activation can also be detected and / or measured using the methods described and exemplified in the working examples. Assays using these or other types of suitable candidate antibodies capable of inhibiting C5 activation can be screened. In certain modalities, inhibition of C5 activation includes at least a decrease of 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% or more in C5 activation in an assay when compared with the effect of a negative control under similar conditions.In some modalities, it refers to an activation inhibition of at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% or more. D. Immunoconjugates

[00287] The invention also provides immunoconjugates comprising an anti-C5 antibody conjugated with one or more cytotoxic agents, such as chemotherapeutic agents or drugs, growth-inhibiting agents, toxins (e.g., protein toxins, enzymatically active toxins from bacteria, fungi, plants or of animal origin or fragments thereof) or radioactive isotopes.

[00288] In one embodiment, an immunoconjugate is an antibody-drug conjugate (ADC) in which an antibody is conjugated to one or more drugs including, but not limited to, a maytansinoid (see U.S. Patents Nos. 5,208,020, 5,416,064 and European Patent EP 0 425 235 B1); an auristatin, such as the portions Petition 870260079322, dated 07 / 08 / 2026, page 117 / 523 109 / 167 of the drug monomethylauristatin DE and DF (MMAE and MMAF) (see U.S. Patents Nos. 5,635,483 and 5,780,588 and 7,498,298); a dolastatin; a calicheamicin or derivative thereof (see U.S. Patents Nos. 5,712,374, 5,714,586, 5,739,116, 5,767,285, 5,770,701, 5,770,710, 5,773,001 and 5,877,296; Hinman et al., Cancer Res. 53: 3336-3342 (1993); and Lode et al., Cancer Res. 58: 2925-2928 (1998)); an anthracycline, such as daunomycin or doxorubicin (see Kratz et al., Current Med. Chem. 13: 477-523 (2006); Jeffrey et al., Bioorganic & Med. Chem. Letters 16: 358-362 (2006); Torgov et al., Bioconj. Chem. 16: 717-721 (2005); Nagy et al. al., Proc. Natl. Sci. USA 97: 829-834 (2000); 6,630,579); methotrexate; vindesine; a taxane, such as docetaxel, paclitaxel, larotaxel, tesetaxel and ortataxel; a trichothecene; and CC1065.

[00289] In another embodiment, an immunoconjugate comprises an antibody as described herein conjugated to an enzymatically active toxin or fragment thereof including, but not limited to, diphtheria A chain, active non-binding fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modecin A chain, alpha-sarcin, Aleurites fordii proteins, diantin proteins, Phytolacca americana proteins (PAPI, PAPII and PAP-S), normodia carantia inhibitor, curcin, crotin, Saponina officinalis inhibitor, gelonin, mitogellin, restrictocin, fenomycin, enomycin and trichothecenes.

[00290] In another embodiment, an immunoconjugate comprises an antibody as described herein conjugated to a radioactive atom to form a radioconjugate. A variety of radioactive isotopes are available for the production of radioconjugates. Examples include At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212 and other isotopes. Petition 870260079322, dated 07 / 08 / 2026, page 118 / 523 110 / 167 radioactive Lu. When the radioconjugate is used for detection, it may comprise a radioactive atom for scintigraphic studies, for example, tc99m or I123, or a spin tracer for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese, or iron.

[00291] Conjugates of an antibody and a cytotoxic agent can be made using a variety of bifunctional protein coupling agents, such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), bifunctional imidoester derivatives (such as dimethyl adipimidate HCl), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bisazide compounds (such as bis(p-azidobenzoyl)hexanediamine), bisdiazonium derivatives (such as bis-(p-diazoniumbenzoyl)ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science 238: 1098 (1987).Carbon-14 labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for the conjugation of a radionuclide to an antibody. See WO94 / 11026. The ligand may be a cleavable ligand which facilitates the release of a cytotoxic drug into the cell. For example, an acid-labile ligand, a peptidase-sensitive ligand, a photosensitive ligand, a dimethyl ligand, or a disulfide-containing ligand (Chari et al., Cancer Res. 52: 127-131 (1992); U.S. Patent No. 5,208,020) may be used.

[00292] Immunoconjugates or ADCs expressly consider Petition 870260079322, dated 07 / 08 / 2026, page 119 / 523 111 / 167 of those listed herein include, but are not limited to, those conjugates prepared with crosslinking reagents including, but not limited to, BMPs, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC and sulfo-SMPB and SVSB (succinimidyl-(4-vinylsulfone)benzoate), which are commercially available (e.g., from Pierce Biotechnology, Inc., Rockford, IL., USA). E. Methods and Compositions for Diagnosis and Detection

[00293] In certain embodiments, any of the anti-C5 antibodies provided herein is useful for detecting the presence of C5 in a biological sample. The term "detect," as used herein, encompasses quantitative or qualitative detection. In certain embodiments, a biological sample comprises a cell or tissue, such as serum, whole blood, plasma, biopsy sample, tissue sample, cell suspension, saliva, sputum, oral fluid, cerebrospinal fluid, amniotic fluid, ascitic fluid, milk, colostrum, mammary gland secretion, lymph, urine, sweat, lacrimal fluid, gastric fluid, synovial fluid, peritoneal fluid, lens fluid, and mucus.

[00294] In one embodiment, an anti-C5 antibody is provided for use in a diagnostic or detection method. In a further aspect, a method for detecting the presence of C5 in a biological sample is provided. In certain embodiments, the method comprises contacting the biological sample with an anti-C5 antibody as described herein under conditions that allow the anti-C5 antibody to bind to C5 and detecting whether a complex is formed between the anti-C5 antibody and C5. Such a method may be an in vitro or in vivo method. In one embodiment, an anti-C5 antibody is used to select individuals eligible for therapy with an anti-C5 antibody, for example, where C5 is a biomarker for patient selection.

[00295] In another modality, a method is provided for selecting Petition 870260079322, dated 07 / 08 / 2026, page 120 / 523 112 / 167 to identify an individual who has a complement-mediated disease or condition involving excessive or uncontrolled C5 activation as suitable for a therapy comprising an anti-C5 antibody of the present invention. In certain embodiments, the method comprises (a) detecting a genetic variation in C5 derived from the individual and (b) selecting the individual as suitable for therapy comprising an anti-C5 antibody of the present invention when the genetic variation is detected in C5 derived from the individual. In another embodiment, a method is provided for selecting a therapy for an individual who has a complement-mediated disease or condition involving excessive or uncontrolled C5 activation.In certain embodiments, the method comprises (a) detecting a genetic variation in C5 derived from the individual and (b) selecting a therapy comprising an anti-C5 antibody of the present invention for the individual when the genetic variation is detected in C5 derived from the individual.

[00296] In another embodiment, a method is provided for treating an individual who has a complement-mediated disease or condition involving excessive or uncontrolled C5 activation. In certain embodiments, the method comprises (a) detecting a genetic variation in C5 derived from the individual, (b) selecting the individual as suitable for therapy comprising an anti-C5 antibody of the present invention when the genetic variation is detected in C5 derived from the individual, and (c) administering an anti-C5 antibody of the present invention to the individual.

[00297] In another embodiment, an anti-C5 antibody of the present invention is provided for use in the treatment of an individual who has a complement-mediated disease or condition involving excessive or uncontrolled C5 activation. In certain embodiments, the individual is treated with an anti-C5 antibody of the present invention. Petition 870260079322, dated 07 / 08 / 2026, page 121 / 523 113 / 167 Convention when genetic variation is detected in the C5 derived from the individual.

[00298] In another embodiment, in vitro use of a genetic variation in C5 is provided to select an individual who has a complement-mediated disease or condition involving excessive or uncontrolled activation of C5 as suitable for a therapy comprising an anti-C5 antibody of the present invention. In certain embodiments, the individual is selected as suitable for therapy when the genetic variation is detected in the C5 derived from the individual. In another embodiment, in vitro use of a genetic variation in C5 is provided to select a therapy for an individual who has a complement-mediated disease or condition involving excessive or uncontrolled activation of C5. In certain embodiments, a therapy comprising an anti-C5 antibody of the present invention is selected for the individual when the genetic variation is detected in the C5 derived from the individual.

[00299] It has been reported that some patients who have genetic variation in C5 exhibit a poor response to therapy comprising an existing anti-C5 antibody (Nishimura et al., N. Engl. J. Med. 370: 632-639 (2014)). It is recommended that such a patient be treated with therapy comprising an anti-C5 antibody of the present invention, since such an antibody has an inhibitory activity against the activation of C5 variants as well as wild-type C5, as demonstrated in the working examples below.

[00300] The detection of a genetic variation in C5 can be performed using a method known in the state of the art. Such a method may include sequencing, PCR, RT-PCR, and a hybridization-based method, such as Southern blot or Northern blot, but without limitation to these. C5 variants may comprise at least one genetic variation. The genetic variation may be selected Petition 870260079322, dated 07 / 08 / 2026, page 122 / 523 114 / 167 nothing of a group consisting of V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N and E1437D. Here, R885H, for example, signifies a genetic variation in which arginine at position 885 is replaced by histidine. In certain embodiments, a C5 variant has biological activity similar to that of wild-type C5.

[00301] Exemplary disorders that can be diagnosed using an antibody of the invention include rheumatoid arthritis (RA); systemic lupus erythematosus (SLE); lupus disease; ischemia-reperfusion injury (IRI); asthma; paroxysmal nocturnal hemoglobinuria (PNH); hemolytic uremic syndrome (HUS) (e.g., atypical hemolytic uremic syndrome (aHUS)); dense deposit disease (DDD); neuromyelitis optica (NMO); multifocal motor neuropathy (MMN); multiple sclerosis (MS); cystic sclerosis; macular degeneration (e.g., age-related macular degeneration (AMD)); hemolysis, hemolysis syndrome due to elevated liver enzymes and low platelet count (HELLP); thrombotic thrombocytopenic purpura (TTP); spontaneous fetal loss; epidermolysis bullosa; recurrent fetal loss; pre-eclampsia; head trauma; myasthenia gravis;Cold agglutinin disease; Sjögren's syndrome; dermatomyositis; bullous pemphigoid; phototoxic reactions; E. coli Shiga toxin-related hemolytic uremic syndrome; typical or infectious hemolytic uremic syndrome (tHUS); C3 glomerulonephritis; antineutrophil cytoplasmic antibody-associated vasculitis (ANCA); humoral and vascular transplant rejection; acute antibody-mediated rejection (AMR); graft dysfunction; myocardial infarction; allogeneic transplantation; sepsis; coronary artery disease; hereditary angioedema; dermatomyositis; Graves' disease; atherosclerosis; mal de; Petition 870260079322, dated 07 / 08 / 2026, page 123 / 523 115 / 167 Alzheimer's disease (AD); Huntington's disease; Creutzfeldt-Jakob disease; Parkinson's disease; cancers; wounds; septic shock; spinal cord injury; uveitis; diabetic eye diseases; retinopathy of prematurity; glomerulonephritis; membranous nephritis; immunoglobulin A nephropathy; adult respiratory distress syndrome (ARDS); chronic obstructive pulmonary disease (COPD); cystic fibrosis; hemolytic anemia; paroxysmal cold hemoglobinuria; anaphylactic shock; allergy; osteoporosis; osteoarthritis; Hashimoto's thyroiditis; type I diabetes; psoriasis; pemphigus; autoimmune hemolytic anemia (AIHA); idiopathic thrombocytopenic purpura (ITP); Goodpasture syndrome; Degos disease; antiphospholipid syndrome (APS); catastrophic APS (CAPS); a cardiovascular disorder; myocarditis; a cerebrovascular disorder; a peripheral vascular disorder; a renovascular disorder;a mesenteric / enteric vascular disorder; vasculitis; Henoch-Schönlein purpura nephritis; Takayasu disease; dilated cardiomyopathy; diabetic angiopathy; Kawasaki disease (arteritis); venous gas embolism (VGE); restenosis after stent placement; rotational atherectomy; membranous nephropathy; Guillain-Barré syndrome (GBS); Fisher syndrome; antigen-induced arthritis; synovial inflammation; viral infections; bacterial infections; fungal infections; and injury resulting from myocardial infarction, cardiopulmonary bypass, and hemodialysis.

[00302] In certain embodiments, labeled anti-C5 antibodies are provided. Labels include, but are not limited to, labels or portions that are directly detected (such as fluorescent, chromophoric, electron-dense, chemiluminescent and radioactive labels), as well as portions such as enzymes or ligands that are detected indirectly, for example, through an enzymatic reaction or molecular interaction. Exemplificative markers Petition 870260079322, dated 07 / 08 / 2026, page 124 / 523 116 / 167 include, but are not limited to, the radioisotopes 32P, 14C, 125I, 3H and 131I, fluorophores such as rare earth chelates or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, luciferase, for example, firefly luciferase and bacterial luciferase (United States Patent No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, horseradish peroxidase (HRP), alkaline phosphatase, beta-galactosidase, glucoamylase, lysozyme, saccharide oxidases, for example, glucose oxidase, galactose oxidase and glucose-6-phosphate dehydrogenase, heterocyclic oxidases such as uricase and xanthine oxidase, coupled with a An enzyme that uses hydrogen peroxide to oxidize a dye precursor such as HRP, lactoperoxidase or microperoxidase, biotin / avidin, spin markers, bacteriophage markers, stable free radicals, and so on. F. Pharmaceutical Formulations

[00303] Pharmaceutical formulations of an anti-C5 antibody as described herein are prepared by mixing such antibody having the desired degree of purity with one or more optional pharmaceutically acceptable carriers (Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions.Pharmaceutically acceptable carriers are generally non-toxic to receptors at the dosages and concentrations used and include, but are not limited to: 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 or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexane; 3-pentanol; and m-cresol); low molecular weight polypeptides (less than about 10 residues); proteins such as serum albumin, gelatin or immunoglobulins; and high-density polymers. Petition 870260079322, dated 07 / 08 / 2026, page 125 / 523 117 / 167 drophilic substances, 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 counter-ions, such as sodium; metal complexes, for example, Zn-protein complexes; and / or nonionic surfactants, such as polyethylene glycol (PEG). Pharmaceutically acceptable exemplary carriers also include interstitial drug dispersing agents, such as soluble neutral active hyaluronidase glycoproteins (sHASEGP), for example, soluble human hyaluronidase PH-20 glycoproteins, such as rHuPH20 (HYLENEX (registered trademark), Baxter International, Inc.). Certain exemplary sHASEGPs and methods of use, including rHuPH20, are described in U.S. Publications Nos. 2005 / 0260186 and 2006 / 0104968.In one aspect, sHASEGP is combined with one or more additional glycosaminoglycanases, such as chondroitinases.

[00304] Exemplary lyophilized antibody formulations are described in U.S. Patent No. 6,267,958. Aqueous antibody formulations include those described in U.S. Patent No. 6,171,586 and in document WO2006 / 044908, the latter formulations including a histidine-acetate buffer.

[00305] The formulation may also contain more than one active ingredient as needed for the particular indication to be treated, preferably those with complementary activities that do not adversely affect each other. Such active ingredients are appropriately present in combinations in amounts that are effective for the intended purpose.

[00306] The active ingredients can be encapsulated in microcapsules prepared, for example, using coacerva techniques. Petition 870260079322, dated 07 / 08 / 2026, page 126 / 523 118 / 167 interfacial tion or polymerization, for example, hydroxymethylcellulose or gelatin microcapsules and (poly(methylmethacrylate)) microcapsules, respectively, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such techniques are described in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980).

[00307] Extended-release preparations may be prepared. Suitable examples of extended-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, the matrices of which are in the form of shaped articles, for example, films or microcapsules.

[00308] The formulations to be used for in vivo administration are generally sterile. Sterilization can be easily achieved, for example, by filtration through sterile filtration membranes. G. Therapeutic Methods and Compositions

[00309] Any of the anti-C5 antibodies provided here can be used in therapeutic methods.

[00310] In one aspect, an anti-C5 antibody for use as a medicament is provided. In further aspects, an anti-C5 antibody for use in the treatment of a complement-mediated disease or condition involving excessive or uncontrolled C5 activation is provided. In certain embodiments, an anti-C5 antibody for use in a treatment method is provided. In certain embodiments, the invention provides an anti-C5 antibody for use in the treatment method of an individual who has a complement-mediated disease or condition involving excessive or uncontrolled C5 activation, comprising administering an effective amount of the anti-C5 antibody to the individual. In one such embodiment Petition 870260079322, dated 07 / 08 / 2026, page 127 / 523 119 / 167 modalities, the method still comprises administering to the individual an effective quantity of at least one additional therapeutic agent. An individual, according to any of the above modalities, is preferably a human being.

[00311] When the antigen is a soluble protein, the binding of an antibody to its antigen can result in a prolonged half-life of the antigen in plasma (i.e., reduced clearance of the antigen from plasma), since the antibody itself has a longer half-life in plasma and serves as a carrier for the antigen. This is due to the recycling of the antigen-antibody complex by FcRn via the endosomal pathway into cells (Roopenian, Nat. Rev. Immunol. 7 (9): 715-725 (2007)). However, an antibody with pH-dependent binding characteristics, which binds to its antigen in a neutral extracellular environment while releasing it into acidic endosomal compartments after entering cells, is expected to have superior antigen neutralization and elimination properties compared to its pH-independent counterpart (Igawa et al., Nat. Biotech. 28 (11): 1203-1207 (2010); Devanaboyina et al., mAbs 5 (6): 851-859 (2013); document WO2009 / 125825).

[00312] In other embodiments, the invention provides an anti-C5 antibody for use in increasing the clearance of C5 from plasma. In certain embodiments, the invention provides an anti-C5 antibody for use in a method for increasing the clearance of C5 from plasma in an individual comprising administering to the individual an effective amount of the anti-C5 antibody to increase the clearance of C5 from plasma. In one embodiment, an anti-C5 antibody increases the clearance of C5 from plasma when compared with a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. An individual, according to any of the modes Petition 870260079322, dated 07 / 08 / 2026, page 128 / 523 120 / 167 of the above is, preferably, a human being.

[00313] In other embodiments, the invention provides an anti-C5 antibody for use in suppressing the accumulation of C5 in plasma. In certain embodiments, the invention provides an anti-C5 antibody for use in a method for suppressing the accumulation of C5 in plasma in an individual comprising administering to the individual an effective amount of the anti-C5 antibody to suppress the accumulation of C5 in plasma. In one embodiment, the accumulation of C5 in plasma is the result of the formation of an antigen-antibody complex. In another embodiment, an anti-C5 antibody suppresses the accumulation of C5 in plasma when compared to a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. An individual, according to either of the above embodiments, is preferably a human being.

[00314] An anti-C5 antibody of the present invention can inhibit C5 activation. In other embodiments, the invention provides an anti-C5 antibody for use in inhibiting C5 activation. In certain embodiments, the invention provides an anti-C5 antibody for use in a method of inhibiting C5 activation in an individual comprising administering to the individual an effective amount of the anti-C5 antibody to inhibit C5 activation. In one embodiment, C5-mediated cytotoxicity is suppressed by inhibiting C5 activation. An individual, according to any of the above embodiments, is preferably a human being.

[00315] In a further aspect, the invention provides the use of an anti-C5 antibody in the manufacture or preparation of a medicament. In one embodiment, the medicament is for the treatment of a complement-mediated disease or condition which involves excessive or uncontrolled activation of C5. In a further embodiment, the medicament is for use in a method of treating Petition 870260079322, dated 07 / 08 / 2026, page 129 / 523 121 / 167 a complement-mediated disease or condition involving excessive or uncontrolled C5 activation comprising administering an effective amount of the drug to an individual who has the complement-mediated disease or condition involving excessive or uncontrolled C5 activation. In one such embodiment, the method further comprises administering an effective amount of at least one additional therapeutic agent to the individual. An individual, according to any of the above embodiments, is preferably a human being.

[00316] In another embodiment, the drug is for enhancing the elimination of C5 from plasma. In a further embodiment, the drug is for use in a method for enhancing the elimination of C5 from plasma in an individual comprising administering to the individual an effective amount of the drug to enhance the elimination of C5 from plasma. In one embodiment, an anti-C5 antibody enhances the elimination of C5 from plasma when compared with a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. An individual, according to any of the above embodiments, may be a human being.

[00317] In another embodiment, the drug is for suppressing the accumulation of C5 in plasma. In another embodiment, the drug is for use in a method for suppressing the accumulation of C5 in plasma in an individual comprising administering to the individual an effective amount of the drug to suppress the accumulation of C5 in plasma. In one embodiment, the accumulation of C5 in plasma is a result of the formation of an antigen-antibody complex. In another embodiment, an anti-C5 antibody suppresses the accumulation of C5 in plasma when compared with a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. An individual, according to either of the above embodiments, may be a human being. Petition 870260079322, dated 07 / 08 / 2026, page 130 / 523 122 / 167

[00318] An anti-C5 antibody of the present invention can inhibit C5 activation. In another embodiment, the drug is for inhibiting C5 activation. In a further embodiment, the drug is for use in a method of inhibiting C5 activation in an individual comprising administering to the individual an effective amount of the drug to inhibit C5 activation. In one embodiment, C5-mediated cytotoxicity is suppressed by inhibiting C5 activation. An individual, according to any of the above embodiments, can be a human being.

[00319] In a further aspect, the invention provides a method of treating a complement-mediated disease or condition involving excessive or uncontrolled C5 activation. In one embodiment, the method comprises administering to an individual who has such a complement-mediated disease or condition involving excessive or uncontrolled C5 activation an effective amount of anti-C5 antibody. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent. An individual according to any of the above embodiments may be a human being.

[00320] In another aspect, the invention provides a method for increasing the clearance of C5 from plasma in an individual. In one embodiment, the method comprises administering to the individual an effective amount of an anti-C5 antibody to increase the clearance of C5 from plasma. In one embodiment, an anti-C5 antibody increases the clearance of C5 from plasma when compared with a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. In one embodiment, an individual is a human being.

[00321] In another aspect, the invention provides a method for suppressing the accumulation of C5 in the plasma of an individual. In one embodiment, the method comprises administering to the individual a quantity Petition 870260079322, dated 07 / 08 / 2026, page 131 / 523 123 / 167 of the effectiveness of an anti-C5 antibody in suppressing C5 accumulation in plasma. In one embodiment, C5 accumulation in plasma is a result of the formation of an antigen-antibody complex. In another embodiment, an anti-C5 antibody suppresses C5 accumulation in plasma when compared to a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. In one embodiment, an individual is a human being.

[00322] An anti-C5 antibody of the present invention can inhibit C5 activation. In another aspect, the invention provides a method for inhibiting C5 activation in an individual. In one embodiment, the method comprises administering to the individual an effective amount of an anti-C5 antibody to inhibit C5 activation. In one embodiment, C5-mediated cytotoxicity is suppressed by inhibiting C5 activation. In one embodiment, an individual is a human being.

[00323] In a further aspect, the invention provides pharmaceutical formulations comprising any of the anti-C5 antibodies provided herein, for example for use in any of the therapeutic methods above. In one embodiment, a pharmaceutical formulation comprises any of the anti-C5 antibodies provided herein and a pharmaceutically acceptable carrier. In another embodiment, a pharmaceutical formulation comprises any of the anti-C5 antibodies provided herein and at least one additional therapeutic agent.

[00324] In another aspect, the pharmaceutical formulation is for the treatment of a complement-mediated disease or condition involving excessive or uncontrolled C5 activation. In another embodiment, the pharmaceutical formulation is to increase the clearance of C5 from plasma. In one embodiment, an anti-C5 antibody increases the clearance of C5 from plasma when compared to a conventional anti-C5 antibody that does not have dependent binding characteristics. Petition 870260079322, dated 07 / 08 / 2026, page 132 / 523 124 / 167 pH. In another embodiment, the pharmaceutical formulation is for suppressing the accumulation of C5 in plasma. In one embodiment, the accumulation of C5 in plasma is a result of the formation of an antigen-antibody complex. In another embodiment, an anti-C5 antibody suppresses the accumulation of C5 in plasma when compared to a conventional anti-C5 antibody that does not have pH-dependent binding characteristics. An anti-C5 antibody of the present invention can inhibit C5 activation. In another embodiment, the pharmaceutical formulation is for inhibiting C5 activation. In one embodiment, C5-mediated cytotoxicity is suppressed by inhibiting C5 activation. In one embodiment, the pharmaceutical formulation is administered to an individual who has a complement-mediated disease or condition involving excessive or uncontrolled C5 activation. An individual, according to any of the above embodiments, is preferably a human being.

[00325] In one aspect, an individual has wild-type C5. In another aspect, an individual has a variant of C5. In certain embodiments, a variant of C5 has biological activity similar to that of wild-type C5. Such a variant of C5 may comprise at least one selected variation of the group consisting of V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N, and E1437D. Here, R885H, for example, means a genetic variation in which arginine at position 885 is replaced by histidine.

[00326] In a further aspect, the invention provides methods for the preparation of a medicament or a pharmaceutical formulation comprising administering any of the anti-C5 antibodies provided herein with a pharmaceutically acceptable vehicle, for example, for use in any of the therapeutic methods above. In one embodiment, the methods for the preparation of a medicament or a pharmaceutical formulation comprise Petition 870260079322, dated 07 / 08 / 2026, page 133 / 523 125 / 167 still add at least one additional therapeutic agent to the drug or pharmaceutical formulation.

[00327] In certain modalities, the complement-mediated disease or condition involving excessive or uncontrolled C5 activation is selected from the group consisting of rheumatoid arthritis (RA); systemic lupus erythematosus (SLE); lupus disease; ischemia-reperfusion injury (IRI); asthma; paroxysmal nocturnal hemoglobinuria (PNH); hemolytic uremic syndrome (HUS) (e.g., atypical hemolytic uremic syndrome (aHUS)); dense deposit disease (DDD); neuromyelitis optica (NMO); multifocal motor neuropathy (MMN); multiple sclerosis (MS); cystic sclerosis; macular degeneration (e.g., age-related macular degeneration (AMD)); Hemolysis, hemolytic syndrome due to elevated liver enzymes and low platelet count (HELLP); thrombotic thrombocytopenic purpura (TTP); spontaneous fetal loss;Epidermolysis bullosa; recurrent fetal loss; pre-eclampsia; head trauma; myasthenia gravis; cold agglutinin disease; Sjögren's syndrome; dermatomyositis; bullous pemphigoid; phototoxic reactions; E. coli Shiga toxin-related hemolytic uremic syndrome; typical or infectious hemolytic uremic syndrome (tHUS); C3 glomerulonephritis; antineutrophil cytoplasmic antibody-associated vasculitis (ANCA); humoral and vascular transplant rejection; acute antibody-mediated rejection (AMR); graft dysfunction; myocardial infarction; allogeneic transplantation; sepsis; coronary artery disease; hereditary angioedema; dermatomyositis; Graves' disease; atherosclerosis; Alzheimer's disease (AD); Huntington's disease; Creutzfeldt-Jakob disease; Parkinson's disease; cancers; wounds; septic shock; spinal cord injury; Petition 870260079322, dated 07 / 08 / 2026, page 134 / 523 126 / 167 spinal; uveitis; diabetic eye diseases; retinopathy of prematurity; glomerulonephritis; membranous nephritis; immunoglobulin A nephropathy; adult respiratory distress syndrome (ARDS); chronic obstructive pulmonary disease (COPD); cystic fibrosis; hemolytic anemia; paroxysmal cold hemoglobinuria; anaphylactic shock; allergy; osteoporosis; osteoarthritis; Hashimoto's thyroiditis; type I diabetes; psoriasis; pemphigus; autoimmune hemolytic anemia (AIHA); idiopathic thrombocytopenic purpura (ITP); Goodpasture's syndrome; Degos disease; antiphospholipid syndrome (APS); catastrophic APS (CAPS); a cardiovascular disorder; myocarditis; a cerebrovascular disorder; a peripheral vascular disorder; a renovascular disorder; a mesenteric / enteric vascular disorder; vasculitis; Henoch-Schönlein purpura nephritis; Takayasu's disease; dilated cardiomyopathy; diabetic angiopathy;Kawasaki disease (arteritis); venous gas embolism (VGE); restenosis after stent placement; rotational atherectomy; membranous nephropathy; Guillain-Barré syndrome (GBS); Fisher syndrome; antigen-induced arthritis; synovial inflammation; viral infections; bacterial infections; fungal infections; and injury resulting from myocardial infarction, cardiopulmonary bypass, and hemodialysis.

[00328] In certain modalities, the complement-mediated disease or condition is an ocular pathological condition. In other modalities, the ocular condition is macular degeneration. In other modalities, macular degeneration is AMD. In other modalities, AMD is the dry form of AMD.

[00329] In certain modalities, the complement-mediated disease or condition is PNH.

[00330] In certain modalities, the complement-mediated disease or condition is a myocardial infarction. Petition 870260079322, dated 07 / 08 / 2026, page 135 / 523 127 / 167

[00331] In certain modalities, the complement-mediated disease or condition is RA.

[00332] In certain modalities, the complement-mediated disease or condition is osteoporosis or osteoarthritis.

[00333] In certain modalities, the complement-mediated disease or condition is inflammation.

[00334] In certain modalities, the complement-mediated disease or condition is cancer.

[00335] The antibodies of the invention can be used alone or in combination with other agents in a therapy. For example, an antibody of the invention can be co-administered with at least one additional therapeutic agent.

[00336] The combination therapies noted above encompass combined administration (where two or more therapeutic agents are included in the same or separate formulations) and separate administration, in which case the administration of the antibody of the invention may occur before, simultaneously with, and / or after the administration of the additional therapeutic agent or agents. In one embodiment, the administration of the anti-C5 antibody and the administration of an additional therapeutic agent occur within about one month or within one, two, or three weeks or within about one, two, three, four, five, or six days of each other.

[00337] An antibody of the invention (and any additional therapeutic agent) can be administered via any suitable means, including parenteral, intrapulmonary and intranasal administration and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal or subcutaneous administration. Dosing can be via any suitable route, for example, injections, such as intravenous or subcutaneous injections, depending in part on whether the Petition 870260079322, dated 07 / 08 / 2026, page 136 / 523 128 / 167 administration is for short-term or chronic use. Various dosing regimens, including but not limited to single or multiple administrations over several time points, bolus administration, and pulsed infusion, are contemplated herein.

[00338] The antibodies of the invention will be formulated, dosed, and administered in a manner consistent with good human practice. Factors for consideration in this context include the particular disorder to be treated, the particular mammal to be treated, the clinical condition of the individual patient, the cause of the disorder, the site of agent release, the method of administration, the administration schedule, and other factors known to physicians. The antibody need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of antibody present in the formulation, the type of disorder or treatment, and other factors discussed above.These are generally used at the dosages above and with routes of administration as described herein, or between approximately 1% and 99% of the dosages described herein, or at any dosage and by any route that is determined empirically / clinically to be appropriate.

[00339] For the prevention or treatment of a disease, the appropriate dosage of an antibody of the invention (when used alone or in combination with one or more additional therapeutic agents) will depend on the type of disease to be treated, the type of antibody, the severity and course of the disease, whether the antibody is administered for preventive or therapeutic purposes, prior therapy, the patient's medical history and response to the antibody, and at the discretion of the attending physician. The antibody is appropriately administered to the patient once or during a series of treatments. Depending on the type and severity of the disease, approximately Petition 870260079322, dated 07 / 08 / 2026, p. 137 / 523 A dosage of 129 / 167 micrograms / kg to 15 mg / kg (e.g., 0.1 mg / kg to 10 mg / kg) of antibody may be the initial candidate dosage for administration to the patient, for example, in one or more administrations or via continuous infusion. A typical daily dosage will range from about 1 microgram / kg to 100 mg / kg or more, depending on the factors mentioned above. For repeated administrations over several days or more, depending on the condition, treatment will be maintained until the desired suppression of disease symptoms occurs. An exemplary antibody dosage will be in the range of about 0.05 mg / kg to about 10 mg / kg. Therefore, one or more doses of about 0.5 mg / kg, 2.0 mg / kg, 4.0 mg / kg, or 10 mg / kg (or any combination thereof) may be administered to the patient.These doses can be administered intermittently, for example, every week or every three weeks (for example, so that the patient receives between about two and about twenty, or for example, about six doses of the antibody). A larger initial dose, followed by one or more smaller doses, may be administered. The progress of this therapy is easily monitored using conventional techniques and assays.

[00340] It should be understood that any of the above formulations or therapeutic methods can be performed using an immunoconjugate of the invention in place of or in addition to an anti-C5 antibody. H. Manufactured Articles

[00341] In another aspect of the invention, a manufactured article is provided containing materials useful for the treatment, prevention and / or diagnosis of the disorders described above. The manufactured article comprises a container and a label or leaflet associated with the container. Suitable containers include, for example, glass bottles, vials, syringes, IV solution bags, etc. The containers may Petition 870260079322, dated 07 / 08 / 2026, page 138 / 523 130 / 167 can be formed from a variety of materials such as glass or plastic. The container holds a composition that is alone or combined with another composition effective for the treatment, prevention and / or diagnosis of the condition and may have a sterile access port (for example, the container may be an intravenous solution bag or a bottle with a cap that can be pierced by the needle of a hypodermic syringe). At least one active agent in the composition is an antibody of the invention. The label or package insert indicates that the composition is used for the treatment of the condition of choice. In addition, the manufactured article may comprise (a) a first container with a composition contained therein, wherein the composition comprises an antibody of the invention; and (b) a second container with a composition contained therein, wherein the composition comprises an additional cytotoxic or therapeutic agent.The manufactured article in this embodiment of the invention may further comprise a package insert indicating that the composition can be used to treat a particular condition. Alternatively or additionally, the manufactured article may also comprise a second (or third) container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may also include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[00342] It should be understood that any of the above manufactured articles may include an immunoconjugate of the invention in place of or in addition to an anti-C5 antibody. EXAMPLES

[00343] The following are examples of methods and compositions of the invention. It should be understood that various other embodiments Petition 870260079322, dated 07 / 08 / 2026, page 139 / 523 131 / 167 can be practiced, given the general description provided above. Example 1 C5 preparation 1.1 Expression and purification of recombinant human and cynomolgus monkey C5

[00344] Recombinant human C5 (NCBI GenBank accession number: NP_001726.2, SEQ ID NO: 39) was transiently expressed using the FreeStyle293-F cell line (Thermo Fisher, Carlsbad, CA, USA). Conditioned medium expressing C5 was diluted with an equal volume of milliQ water, then applied to a Q-sepharose FF or Q-sepharose HP anion exchange column (GE healthcare, Uppsala, Sweden), followed by elution with a NaCl gradient. The fractions containing human C5 were pooled, and then the salt concentration and pH were adjusted to 80 mM NaCl and pH 6.4, respectively. The resulting sample was applied to an SP-sepharose HP cation exchange column (GE healthcare, Uppsala, Sweden) and eluted with a NaCl gradient. Fractions containing human C5 were pooled and subjected to a Hydroxyapatite CHT ceramic column (Bio-Rad Laboratories, Hercules, CA, USA).The human C5 eluate was then applied to a Superdex 200 gel filtration column (GE healthcare, Uppsala, Sweden). The fractions containing human C5 were pooled and stored at -150 degrees C.

[00345] The expression and purification of recombinant C5 from cynomolgus monkey (NCBI GenBank accession number: XP_005580972, : SEQ ID NO: 44) was performed in the same manner as the human counterpart. 1.2 Purification of cynomolgus monkey C5 (cyno C5) plasma

[00346] Cynomolgus monkey plasma samples were applied to SSL7-agarose (Invivogen, San Diego, CA, USA) followed by elution Petition 870260079322, dated 07 / 08 / 2026, page 140 / 523 132 / 167 with 100 mM sodium acetate, pH 3.5. The fractions containing cynoC5 were immediately neutralized and subjected to a Protein A HP column (GE healthcare, Uppsala, Sweden) in tandem with Peptide M agarose (Invivogen, San Diego, CA, USA). The flow through the fraction was then applied to a Superdex 200 gel filtration column (GE healthcare, Uppsala, Sweden). The fractions containing cynoC5 were pooled and stored at -80°C. Example 2 Generation of anti-C5 antibodies 2.1. Antibody screening

[00347] Anti-C5 antibodies were prepared, selected and assayed as follows:

[00348] NZW rabbits aged twelve to sixteen weeks were immunized intradermally with human C5 and / or monkey C5 (50 to 100 micrograms / dose / rabbit). This dose was repeated 4 to 5 times over a period of 2 months. One week after the final immunization, the spleen and blood of the immunized rabbits were collected. Antigen-specific B cells were stained with labeled antigen, sorted with an FCM cell sorter (FACS aria III, BD), and placed in 96-well plates with a density of one cell / well of 25,000 cells / well of EL4 (European Collection of Cell Cultures) cells and activated rabbit T cells in conditioned medium diluted 20 times and cultured for 7 to 12 days. EL4 cells were treated with mitomycin C (Sigma, Cat. No. M4287) for 2 hours and then washed three times. The conditioned medium with activated T cells was prepared by culturing rabbit thymocytes in RPMI-1640 containing Phytohemagglutinin-M (Roche, No.1 1082132-001), 12-myristate 13-phorbol acetate (Sigma, Cat No. P1585) and 2% FBS. After culturing, B cell culture supernatants were collected for analysis. Petition 870260079322, dated 07 / 08 / 2026, page 141 / 523 133 / 167 posterior and the pellets were cryopreserved.

[00349] An ELISA assay was used to test the specificity of antibodies in a B cell culture supernatant. Streptavidin (GeneScript, Cat. No. Z02043) was coated onto a 384-well MAXISorp plate (Nunc, Cat. No. 164688) at 50nM in PBS for 1 hour at room temperature. The plates were then blocked with Blocking One (Nacalai Tesque, Cat. No. 03953-95) diluted 5 times. Human or monkey C5 was labeled with NHSPEG4-Biotin (PIERCE, Cat. No. 21329) and added to the blocked ELISA plates, incubated for 1 hour, and washed. B cell culture supernatants were added to the ELISA plates, incubated for 1 hour, and washed. Binding was detected to goat anti-rabbit peroxidase IgG from Horseradish (BETHYL, Cat. No. A120-111P) followed by the addition of ABTS (KPL, Cat. No. 50-66-06).

[00350] An ELISA assay was used to evaluate the pH-dependent binding of antibodies against C5. Goat anti-rabbit Fc IgG (BETHYL, Cat. No. A120-111A) diluted to 1 microgram / ml with PBS(-) was added to a 384-well MAXISorp plate (Nunc, Cat. No. 164688), incubated for 1 hour at room temperature, and blocked with Blocking One (Nacalai Tesque, Cat. No. 03953-95) diluted 5 times. After incubation, the plates were washed, and B-cell culture supernatants were added. The plates were incubated for 1 hour, washed, and 500 pM of biotinylated human or monkey C5 were added and incubated for 1 hour. After incubation, the plates were washed and incubated with MES buffer pH 7.4 (20 mM MES, 150 mM NaCl, and 1.2 mM CaCl2) or MES buffer pH 5.8 (20 mM MES, 150 mM NaCl, and 1 mM EDTA) for 1 hour at room temperature.After incubation, the binding of biotinylated C5 was detected by the horseradish streptavidin peroxidase conjugate (Thermo Scientific, Cat. No. 21132) followed. Petition 870260079322, dated 07 / 08 / 2026, page 142 / 523 134 / 167 by adding ABTS (KPL, Cat. No. 50-66-06).

[00351] The Octet RED384 system (Pall Life Sciences) was used to evaluate the affinity and pH-dependent binding of antibodies against C5. Antibodies secreted in the supernatant of B cell culture were loaded onto the tip of a Protein A biosensor (Pall Life Sciences) and immersed in 50 nM human or monkey C5 in MES pH 7.4 to analyze the association kinetics. Dissociation kinetics were analyzed in MES buffer pH 7.4 and MES buffer pH 5.8.

[00352] A total of 41,439 B cell lines were screened for affinity and pH-dependent binding to human or monkey C5, and 677 lines were selected and designated as CFA0001-0677. RNA from the selected lines was purified from cryopreserved cell pellets using ZR-96 QuickRNA kits (ZYMO RESEARCH, Cat. No. R1053). DNA encoding the antibody heavy chain variable regions in the selected lines was amplified by reverse transcription PCR and recombined with DNA encoding the F760G4 heavy chain constant region (: SEQ ID NO: 33) or F939G4 heavy chain constant region (: SEQ ID NO: 34). DNA encoding the light chain variable regions was amplified by reverse transcription PCR and recombined with DNA encoding the k0MTC light chain constant region (: SEQ ID NO: 36).Separately, the heavy and light chain genes of an existing humanized anti-C5 antibody, eculizumab (EcuH-G2G4, SEQ ID NO: 29 and EcuL-k0, SEQ ID NO: 30), were synthesized. DNA encoding VH (EcuH, SEQ ID NO: 31) was fused in phase with DNA encoding a modified human IgG4 heavy chain (F760G4, SEQ ID NO: 33), and DNA encoding VL (EcuL, SEQ ID NO: 32) was fused in phase with DNA encoding the constant region of the k0 light chain (SEQ ID NO: 37). Each of the sequences... Petition 870260079322, dated 07 / 08 / 2026, page 143 / 523 Fused 135 / 167 coding cells were also cloned into an expression vector. Antibodies were expressed in FreeStyle™293-F cells (Invitrogen) and purified from the culture supernatant to assess functional activity. Neutralizing antibodies were evaluated by the complement activity inhibition assay using a complement lysis assay as described in Example 5.1.2.2. Epitope compartmentalization by sandwich ELISA

[00353] Anti-C5 antibodies with high affinity, pH dependence, or neutralizing activity were selected for further analysis. A sandwich ELISA assay was used to pool selected antibodies into different epitope bins that bind to the same or overlapping epitopes of the C5 protein. Unlabeled capture antibodies were diluted to 1 microgram / ml with PBS (-) and added to 384-well MAXISorp plates (Nunc, Cat. No. 164688). The plates were incubated for 1 hour at room temperature and blocked with Blocking One (Nacalai Tesque, Cat. No. 03953-95) diluted 5 times. The plates were incubated for 1 hour, washed, and 2 nM of human C5 were added and incubated for 1 hour. After incubation, the plates were washed and labeled detection antibodies (1 microgram / mL, biotinylated with NHS-PEG4Biotin) were added.After 1 hour of incubation, binding of the biotinylated antibody was detected with the streptavidin-horseradish peroxidase conjugate (Thermo Scientific, Cat. No. 21132) followed by the addition of ABTS (KPL, Cat. No. 50-66-06).

[00354] All anti-C5 antibodies were used as both a capture antibody and a detection antibody and comprehensively paired. As shown in Figure 1, mutually competitive antibodies were grouped into 7 epitope bins: CFA0668, CFA0334, and CFA0319 were grouped into epitope A, CFA0647, Petition 870260079322, dated 07 / 08 / 2026, p. 144 / 523 136 / 167 CFA0589, CFA0341, CFA0639, CFA0635, CFA0330, and CFA0318 were clustered under epitope B; CFA0538, CFA0501, CFA0599, CFA0307, ​​CFA0366, CFA0305, CFA0675, CFA0666, and CFA0672 were clustered under epitope C; eculizumab and CFA0322 were clustered under epitope D; CFA0329 was clustered under epitope E; CFA0359 and CFA0217 were clustered under epitope F; and CFA0579, CFA0328, and CFA0272 were clustered under epitope G. Figure 1 shows the epitope binning of some chimeric anti-C5 antibodies. The VH and VL sequences of the anti-C5 antibodies clustered at the C epitope are listed in Table 2. Table 2 Anti-C5 antibodies clustered at the C epitope. : SEQ ID NO: Antibody VH VL HVR-H1 HVR-H2 HVR-H3 HVR-L1 HVR-L2 HVR-L3 CFA0305 1 11 45 55 65 75 85 95 CFA0307 2 12 46 56 66 76 86 96 CFA0366 3 13 47 57 67 77 87 97 CFA0501 4 14 48 58 68 78 88 98 CFA0538 5 15 49 59 69 79 89 99 CFA0599 6 16 50 60 70 80 90 100 CFA0666 7 17 51 61 71 81 91 101 CFA0672 8 18 52 62 72 82 92 102 CFA0675 9 19 53 63 73 83 93 103 2.3. Humanization and optimization

[00355] Humanization of the variable region of some anti-C5 antibodies was performed in order to reduce the potential immunogenicity of the antibodies. Complementarity-determining regions (CDRs) of rabbit anti-C5 antibody were grafted onto homologous structural regions of human antibody (FRs) using a conventional CDR grafting approach (Nature 321: 522-525 (1986)). The genes encoding the humanized VH and VL were synthesized and combined with a modified human IgG4 CH (SG402, : SEQ ID NO: 35) and a human CL (SK1, : SEQ ID NO: 38), respectively, and each of the combined sequences was cloned into an expression vector. Petition 870260079322, dated 07 / 08 / 2026, page 145 / 523 137 / 167

[00356] Several mutations and mutation combinations were examined to identify mutations and mutation combinations that enhance the binding properties of some of the key antibodies. Multiple mutations were also introduced into the humanized variable regions to enhance C5 binding affinity at neutral pH or to reduce C5 binding affinity at neutral pH. One of the optimized variants, 305LO5 (VH, : SEQ ID NO: 10; VL, : SEQ ID NO: 20; HVR-H1, : SEQ ID NO: 54; HVR-H2, : SEQ ID NO: 64; HVR-H3, : SEQ ID NO: 74; HVR-L1, : SEQ ID NO: 84; HVRL2, : SEQ ID NO: 94; and HVR-L3, : SEQ ID NO: 104), was then generated from CFA0305.

[00357] Antibodies were expressed in HEK293 cells cotransfected with a mixture of heavy chain and lead expression vectors and were purified by Protein A. Example 3 Characterization of anti-C5 antibody binding 3.1 Expression and purification of recombinant antibodies

[00358] Recombinant antibodies were transiently expressed using the FreeStyle293-F cell line (Thermo Fisher, Carlsbad, CA, USA). Purification of the antibodies expressed in conditioned medium was performed using a conventional method using protein A. Gel filtration was subsequently conducted if necessary. 3.2. Assessment of pH dependence

[00359] The kinetic parameters of anti-C5 antibodies against recombinant human C5 were evaluated at pH 7.4 and pH 5.8 at 37°C using a BIACORE (registered trademark) T200 instrument (GE Healthcare). ProA / G (Pierce) was immobilized on a CM4 sensor chip using an amine coupling kit (GE Healthcare) according to the settings recommended by GE Healthcare. Antibodies and analytes were diluted in their respective running buffers, ACES. Petition 870260079322, dated 07 / 08 / 2026, page 146 / 523 138 / 167 pH of 7.4 and pH of 5.8 (20 mM ACES, 150 mM NaCl, 1.2 mM CaCl2, 0.05% Tween 20, 0.005% NaNS). Each antibody was captured on the sensor surface by ProA / G. Antibody capture levels were typically 60 to 90 resonance units (RU). Then, recombinant human C5 was injected at concentrations of 10 and 20 nM or 20 and 40 nM followed by dissociation. The surface was regenerated using 25 mM NaOH. The kinetic parameters of both pH conditions were determined by fitting the sensorgrams with a 1:1 binding model using the BIACORE (registered trademark) T200 program, version 2.0 (GE Healthcare). The sensorgrams of all antibodies are shown in Figures 2A and 2B. The association constant (ka), dissociation constant (kd), and binding affinity (KD) of the antibodies are listed in Table 3.All antibodies, except CFA0330 (VH, : SEQ ID NO: 21 and VL, : SEQ ID NO: 25) and CFA0341 (VH, : SEQ ID NO: 22 and VL, : SEQ ID NO: 26) showed a relatively faster dissociation rate at a pH of 5.8 than at a pH of 7.4. Table 3 Kinetic parameters of anti-C5 antibodies under pH conditions of 7.4 and 5.8 Antibody Name pH 7.4 ka kd KD pH 5.8 ka kd KD CFA0305 3.82E+04 5.89E-04 1.54E-08 4.27E+04 1.83E-02 4.30E-07 CFA0307 3.24E+05 2.63E-06 8.13E-09 2.04E+05 3.34E-02 1.64E-07 CFA0366 1.04E+06 9.34E-03 8.99E-09 9.35E+05 7.03E-02 7.52E-08 CFA0501 4.74E+05 1.69E-03 3.56E-09 1.50E+05 2.62E-02 1.74E-07 CFA0538 4.73E+05 1.85E-03 3.91E-09 1.22E+05 3.01E-02 2.46E-07 CFA0599 4.74E+05 2.81E-03 5.93E-09 4.54E+05 3.73E-02 8.21E-08 CFA0666 3.65E+05 6.26E-04 1.71E-09 2.82E+05 9.39E-03 3.33E-08 CFA0672 5023E+05 1.83E-04 3.51E-10 7.11E+04 9.78E-03 1.38E-07 CFA0675 3.83E+05 4.12E-04 1.08E-09 3.89E+05 6.61E-03 1.70E-08 305-LO5 4.48E+05 2.11E-04 4.71E-10 2.03E+06 2.85E-02 1.40E-08 CFA0330 1.66E+06 2.02E-04 1.22E-10 1.22E+06 2.24E-04 1.84E-10 CFA0341 6.28E+05 9.77E-05 1.55E-10 1.24E+06 7.39E-05 5.95E-11 3.3 Evaluation of cross-reactivity

[00360] To observe the cross-reactivity of anti-C5 antibodies Petition 870260079322, dated 07 / 08 / 2026, page 147 / 523 Kinetic analysis was performed using BIACORE (registered trademark) to test anti-C5 antibodies 139 / 167 against human C5 (hC5) and cynomolgus monkey C5 (cynoC5). The assay setup was the same as described in Example 3.2. Recombinant cynoC5 was injected at concentrations of 2, 10, and 50 nM. Kinetic parameters were determined using the same data setup as described in Example 3.2. Binding kinetics and affinity at pH 7.4 are listed in Table 4. The kinetic parameters against hC5 presented in Table 4 are the results from Example 3.2. All anti-C5 antibodies except CFA0672 showed comparable KD against hC5 and cynoC5. The KD of CFA0672 against cynoC5 was 8 times weaker than against hC5. Table 4 Binding kinetics and affinity of anti-C5 antibodies against hC5 and cynoC5 at a pH of 7.4 Antibody Name Affinity Against hC5 ka kd KD Affinity Against cynoC5 ka kd KD CFA0305 3.82E+04 5.89E-04 1.54E-08 1.21E+04 6.70E-04 5.54E-09 CFA0307 3.24E+05 2.63E-06 8.13E-09 2.90E+05 2.23E-03 7.68E-09 CFA0366 1.04E+06 9.34E-03 8.99E-09 5.04E+05 9.04E-03 1.79E-08 CFA0501 4.74E+05 1.69E-03 3.56E-09 2.26E+05 1.56E-03 5.88E-09 CFA0538 4.73E+05 1.85E-03 3.91E-09 3.05E+05 1.66E-03 5.44E-09 CFA0599 4.74E+05 2.81E-03 5.93E-09 5.24E+05 2.35E-03 4.33E-09 CFA0666 3.65E+05 6.26E-04 1.71E-09 3.14E+05 4.93E-04 1.57E-09 CFA0672 5023E+05 1.83E-04 3.51E-10 6.41E+05 1.85E-03 2.88E-09 CFA0675 3.83E+05 4.12E-04 1.08E-09 2.94E+05 3.87E-04 1.29E-09 Example 4 Epitope mapping of anti-C5 antibodies 4.1 Binding of anti-C5 MAbs to C5 beta-chain derived peptides

[00361] Anti-C5 monoclonal antibodies (MAbs) were tested for binding to C5 beta-chain peptides by Western blot analysis. C5 peptides 19-180, 161-340, 321-500, and 481-660, fused to GST-tags (pGEX-4T-1, GE Healthcare Life Sciences, 28-9545-49), were expressed in E. coli (DH5alpha, TOYOBO, DNA-903). E. coli samples were collected after Petition 870260079322, dated 07 / 08 / 2026, page 148 / 523 140 / 167 of the samples were incubated with 1 mM Isopropyl beta-D-1-thiogalactopyranoside (IPTG) for 5 hours at 37°C and centrifuged at 20,000 x g for 1 min to obtain pellets. The pellets were suspended with a sample of buffer solution (2ME+) (Wako, 191-13272) and used for Western blot analysis. The expression of each peptide was confirmed with anti-GST antibody (Abcam, ab9085) (Figure 3). The arrow indicates C5 peptides fused to GST (46-49 kDa). Anti-C5 MAbs: CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672 and CFA0675, linked to 19-180 C5 (Figure 3). 4.2. Expression and purification of the MG1-MG2 (1-225) domain of human C5

[00362] The recombinant MG1-MG2 domain (SEQ ID NO: 43) of the human C5 beta chain was transiently expressed using the FreeStyle293-F cell line (Thermo Fisher, Carlsbad, CA, USA). The conditioned medium expressing the MG1-MG2 domain was diluted with 1 / 2 vol. milliQ water, followed by application to a Q-sepharose FF anion exchange column (GE healthcare, Uppsala, Sweden). The flow through the anion exchange column fraction was adjusted to pH 5.0 and applied to an SP-sepharose cation exchange column (GE healthcare, Uppsala, Sweden) and eluted with a NaCl gradient. Fractions containing the MG1-MG2 domain were collected from the eluent and subsequently filtered through a Superdex 75 gel filtration column (GE healthcare, Uppsala, Sweden) equilibrated with 1x PBS. The fractions containing the MG1-MG2 domain They were then grouped together and stored at -80 degrees C. 4.3. Ability to connect to the MG1-MG2 domain

[00363] The binding capacity of anti-C5 antibodies to the MG1-MG2 domain was measured using the same assay settings as described in Example 3.2, except that measurements were only performed under pH conditions of 7.4. The MG1-MG2 domain Petition 870260079322, dated 07 / 08 / 2026, page 149 / 523 141 / 167 was injected at concentrations of 20 nM and 40 nM. As shown in Figure 4, all antibodies except eculizumab-F760G4 showed an increased binding response, indicating that these antibodies are MG1-MG2 ligands. Eculizumab-F760G4, which is known as an alpha chain ligand, did not show binding to the MG1-MG2 domain. 4.4. Binding of anti-C5 MAbs to peptides derived from the MG1-MG2 domain of C5

[00364] Anti-C5 MAbs were tested for binding to MG1-MG2 domain-derived peptides by Western blot analysis. C5 peptides: 33-124, 45-124, 52-124, 33-111, 33-108, and 45-111 (: SEQ ID NO: 40), fused to GST-tags, were expressed in E. coli. E. coli samples were collected after incubation with 1 mM IPTG for 5 hours at 37°C and centrifuged at 2000 xg for 1 min to obtain pellets. The pellets were suspended with the sample buffer solution (2ME+) and used for Western blot analysis. The expression of C5-derived peptides was confirmed with the anti-GST antibody (Figure 5A). CFA0305 bound only to the 33-124 peptide (Figure 5B). CFA0305 bound to the beta chain of recombinant human C5 (rhC5) (approximately 70 kDa), which was used as a control. Figure 5C summarizes the reaction of anti-C5 MAbs with C5-derived peptides. 4.5. Binding of anti-C5 MAbs to C5 mutants

[00365] As three amino acid residues in the beta chain of C5—E48, D51, and K109—were predicted to be involved in the binding between C5 and anti-C5 MAbs by crystal structure analysis, anti-C5 MAbs were tested for binding to human C5 point mutants by Western blot analysis. C5 point mutants, in which any one of E48, D51, and K109 was substituted for alanine, were expressed in FS293 cells by lipofection. The culture medium Petition 870260079322, dated 07 / 08 / 2026, p. 150 / 523 Sample 142 / 167 was collected 5 days after lipofection and used for Western blot. SDS-PAGE was performed under reducing conditions. The results are shown in Figure 6. Eculizumab bound to the alpha chain of wild-type (WT) C5 and three C5 point mutants, while CFA0305 bound strongly to the beta chain of WT C5, weakly to the E48A mutant of C5, and did not bind to the beta chain of the D51A and K109 mutants of C5, indicating that these 3 amino acid residues are involved in antibody / antigen interactions. Table 5 presents the Western blot analysis of the anti-C5 MAbs (CFA0305, CFA0307, ​​CFA0366, CFA0501, CFA0538, CFA0599, CFA0666, CFA0672, and CFA0675). The anti-C5 MAbs were clustered on the same C epitope, but the binding patterns are subtly different between the antibodies, suggesting that the C5 binding regions to the anti-C5 MAbs are close to each other, but not identical. Table 5 Summary of the reaction of anti-C5 MAbs with C5 mutants WT E48A D51A K109A ECULIZUMABE + + + + CFA0305 + + - - CFA0307 + - - - CFA0366 + - - - CFA0501 + - - - CFA0538 + - - - CFA0599 + - - + CFA0666 + - - + CFA0672 + - - + CFA0675 + + - + 4.6 BIACORE (registered trademark) linkage analysis of anti-C5 antibodies with C5 mutants

[00366] To test whether residues E48, G51, and K109 are indeed involved in antibody / antigen interactions, BIACORE (registered trademark) binding analysis was performed. Three C5 mutants were prepared: E48A, G51A, and K109A, as described in Example 4.5. Culture supernatant samples containing the C5 mutant overexpressed in FS293 cells were prepared in Petition 870260079322, dated 07 / 08 / 2026, page 151 / 523 143 / 167 micrograms / ml of the C5 mutant. For the BIACORE (registered trademark) binding analysis, the sample was diluted with 10x BIACORE (registered trademark) running buffer (ACES pH 7.4, BSA 10 mg / ml, 1 mg / ml carboxymethyl dextran) to a final sample concentration of 4 micrograms / ml of the C5 mutant.

[00367] The interactions of the three C5 mutants with anti-C5 antibodies were evaluated at 37°C using a BIACORE (registered trademark) T200 instrument (GE Healthcare), using the assay condition described in Example 3.2. ACES buffer pH 7.4 containing 10 mg / ml BSA and 1 mg / ml carboxymethyl dextran was used as running buffer. Eculizumab-F760G4 and 305LO5 were captured in different cell streams by the Fc fragment of the mouse anti-human IgG monoclonal antibody (GE Healthcare). Cell stream 2 was used as the reference surface. Wild-type and mutant C5 proteins were injected onto the sensor surface at a concentration of 4 micrograms / ml to interact with the captured antibodies. At the end of each analysis cycle, the sensor surface was regenerated with 3M MgCb. The results were analyzed using the Bia Evaluation program, version 2.0 (GE Healthcare).Reference cell flow curves (cell flow 1) and blank injections of the running buffer were subtracted from cell flow curves with captured antibodies.

[00368] As shown in Figure 7, all three C5 mutants were able to bind to eculizumab with a similar binding profile compared to wild-type C5. For 305LO5, all three mutants showed a lower binding response to 305LO5 compared to wild-type C5. The D51A and K109A mutants reduced C5 binding to 305LO5 to baseline levels. 4.7. Identification of His residues on C5 that contribute to pH-dependent reactions between anti-C5 antibody and C5 Petition 870260079322, dated 07 / 08 / 2026, page 152 / 523 144 / 167

[00369] Crystalline structure analysis revealed that 3 histidine residues on human C5 are located at the antibody / antigen interface. A histidine residue with a typical pKa of approximately 6.0 is known to contribute to pH-dependent protein-protein interactions (Igawa et al., Biochim Biophys Acta 1844(11): 1943-1950 (2014)). To investigate which of the His residues on the antibody / antigen interface contribute to pH-dependent interactions between the anti-C5 antibody and C5, BIACORE (registered trademark) binding analysis was performed. Three human C5 mutants with a single His mutation (H70Y, H72Y, and H110Y) and one mutant with a double His mutation (H70Y + H110Y) were prepared as follows: single His mutants, in which any one of H70, H72, and H110 is replaced by tyrosine, and a double His mutant, in which H70 and H110 are replaced by tyrosine, were expressed in FS293 cells by lipofection.The binding properties of the C5 His mutant to 305LO5, a pH-dependent anti-C5 antibody, were determined by a modified BIACORE assay (trademark) as described in Example 4.6. Briefly, an additional dissociation phase at pH 5.8 was integrated into the BIACORE assay (trademark) immediately after the dissociation phase at pH 7.4, to evaluate the pH-dependent dissociation between the antibody and antigen of the complexes formed at pH 7.4. The dissociation rate at pH 5.8 was determined by processing and fitting data using the Scrubber 2.0 curve fitting program (BioLogic Software).

[00370] As shown in Figure 8, the single His mutation of C5 to H70 or H110 and the double His mutation (H70 + H110) did not affect the binding of C5 to 305LO5 at neutral pH. Meanwhile, the single His mutation to H72 exhibited significant damage to the binding. Petition 870260079322, dated 07 / 08 / 2026, page 153 / 523 145 / 167 of C5 to 305LO5. The dissociation rates at a pH of 5.8 for the C5 His mutants and the C5-wt protein are shown in Table 6. As shown in Table 6, C5-wt showed the fastest dissociation among 305LO5 at a pH of 5.8 among the C5 antigens tested. The single His mutation in H70 exhibited a dissociation rate almost twice as slow at a pH of 5.8, and the single His mutation in His110 resulted in a slightly slower dissociation rate at a pH of 5.8 compared to C5 wt. The double His mutation in both H70 and H110 resulted in a greater effect on pH-dependent binding with a dissociation rate at a pH of 5.8 almost three times slower than C5 wt. Table 6 Dissociation rate value at pH 5.8 for the binding of C5 His mutants to 305LO5 kd antigens (1 / s) C5-wt 1,1E-2 C5-H70Y 5.3E-3 C5-H110Y 9.3E-3 C5-H70Y, H110Y 3.9E-3 Example 5 Inhibitory activity of anti-C5 antibodies on C5 activation 5.1 Inhibition of complement-activated liposome lysis by anti-C5 MAbs

[00371] Anti-C5 MAbs were tested for complement activity inhibition by liposome lysis assay. Thirty µL of normal human serum (6.7%) (Biopredic, SER018) were mixed with 20 µL of diluted MAb in a 96-well plate and incubated on a shaker for 30 min at 25°C. Liposomes sensitized with antibodies against dinitrophenyl (Autokit CH50, Wako, 995-40801) were transferred to each well and the plate was placed on a shaker for 2 min at 25°C. Petition 870260079322, dated 07 / 08 / 2026, page 154 / 523 146 / 167 degrees C. Fifty microliters of substrate solution (Autokit CH50) were added to each well and mixed with stirring for 2 min at 25 degrees C. The final mixture was incubated at 37 degrees C for 40 minutes, and then the OD at 340 nm of the mixture was measured. The percentage of liposome lysis was defined as 100 x [(OD MAb - OD serum and background liposome)] / [(OD without MAb - OD serum and background liposome)]. Figure 9A shows that the anti-C5 MAbs: CFA0305, 0307, ​​0366, 0501, 0538, 0599, 0666, 0672, and 0675, inhibited liposome lysis. Two pH-independent antibodies, CFA0330 and 0341, also inhibited lysis (Figure 9B). 5.2. Inhibition of C5a generation by anti-C5 MAbs

[00372] Anti-C5 MAbs were tested for C5a generation during liposome lysis to confirm that anti-C5 MAbs inhibit C5 cleavage into Ca and C5b. The C5a level in the liposome lysis assay supernatants was quantified using a C5a ELISA kit (R&D systems, DY2037). All MAbs inhibited C5a generation in the supernatants in a dose-dependent manner (Figures 10A and 10B). 5.3. Inhibition of complement-activated hemolysis by anti-C5 MAbs

[00373] Anti-C5 MAbs were tested for inhibition of classical complement activity in a hemolytic assay. Chicken red blood cells (cRBCs) (Innovative research, IC050810) were washed with buffered gelatin / veronal-saline containing 0.5 mM MgCl2 and 0.15 mM CaCl2 (GVB++) (Boston BioProducts, IBB300X) and then sensitized with chicken anti-RBC antibody (Rockland 103-4139) at 1 µg / ml for 15 minutes at 4°C. The cells were then washed with GVB++ and suspended in the same buffer at 5 x 107 cells / ml. In a 96-well round-bottom microtest plate, 50 µl of normal human serum Petition 870260079322, dated 07 / 08 / 2026, page 155 / 523 147 / 167 (20%) (Biopredic, SER019) were mixed with 50 µL of diluted Mab and incubated on a shaker at 37°C for 30 minutes. Sixty µL of the sensitized cRBC suspension were then incubated at 37°C for 30 minutes. After incubation, the plate was centrifuged at 1000 x g for 2 minutes at 4°C. The supernatants (100 µL) were transferred to the wells of a 96-well flat-bottom microtest plate for OD measurement at 415 nm with a reference wavelength of 630 nm. The percentage of hemolysis was defined as 100 x [(OD MAb - OD serum and cRBCs)] / [(OD without MAb - OD serum and background cRBCs)]. Figure 11 shows that the anti-C5 MAbs CFA0305 and 305LO5 inhibited the hemolysis of cRBCs. 5.4. Inhibition of the alternative complement pathway by anti-C5 MAbs

[00374] A hemolytic assay for the alternative pathway was performed similarly to the hemolytic assay for the classical pathway. Blood collected from New Zealand white rabbits (InVivos) was mixed with the same volume of Alsever's solution (Sigma, A3551) and the mixture was used as rabbit RBCs (rRBCs). rRBCs were washed with GVB supplemented with 2 mM MgCb and 10 mM EGTA and suspended in the same buffer at 7 x 10⁸ cells / ml. In a 96-well round-bottom microtest plate, 40 µl of normal human serum (25%) (Biopredic, SER019) were mixed with 40 µl of diluted MAb and incubated on a shaker at 37°C for 30 minutes. Twenty microliters of the rRBC suspension were then added to the wells containing the serum, and the antibody mixture was incubated at 37°C for 60 minutes. After incubation, the plate was centrifuged at 1000 xg for 2 minutes at 4°C.The supernatants (70 µl) were transferred to wells of a 96-well flat-bottom microtest plate for OD measurement at 415 nm with a wavelength of 630 nm. Figure 12 shows that the anti-MAbs... Petition 870260079322, dated 07 / 08 / 2026, p. 156 / 523 148 / 167 C5: CFA0305 and CFA0672 inhibited hemolysis of rRbcs, indicating that these antibodies inhibit alternative complement pathways. Example 6 Pharmacokinetic study of anti-C5 monoclonal antibodies with human C5 in mice 6.1. In vivo assay using C57BL / 6 mice

[00375] The in vivo kinetics of human C5 (Calbiochem) and anti-human C5 antibody were evaluated after administration of human C5 alone or human C5 and anti-human C5 antibody to C57BL / 6 mice (In Vivos or Biological Resource Center, Singapore). A solution of human C5 (0.01 mg / ml) or a mixed solution containing human C5 and anti-human C5 antibody (0.01 mg / ml and 2 mg / ml (CFA0305-F760G4, CFA0307-F760G4, CFA0366-F760G4, CFA0501F760G4, CFA0538-F760G4, CFA0599-F760G4, CFA0666-F760G4, CFA0672-F760G4 and CFA0675-F760G4) or 0.2 mg / ml (CFA0330F760G4 and CFA0341-F760G4), respectively) was administered once at a dose of 10 ml / kg into the tail vein. In this case, the anti-human C5 antibody is present in excess relative to human C5 and, consequently, it is presumed that almost all human C5 is bound to the antibody. Blood was collected at 5 minutes, seven hours, one day, two days, three days, and seven days after administration. The collected blood was immediately centrifuged at 14°C.The plasma was separated at 000 rpm and 4 degrees C for 10 minutes. The separated plasma was stored in a freezer at -80 degrees C before the assay. The anti-human C5 antibodies used are: as described above CFA0305-F760G4, CFA0307-F760G4, CFA0330-F760G4, CFA0341-F760G4, CFA0366-F760G4, CFA0501-F760G4, CFA0538-F760G4, CFA0599-F760G4, CFA0666-F760G4, CFA0672-F760G4 and CFA0675-F760G4. 6.2. Measurement of total human plasma C5 concentration by Petition 870260079322, dated 07 / 08 / 2026, page 157 / 523 149 / 167 electrochemiluminescence assay (ECL)

[00376] The total plasma concentration of human mouse C5 was measured by ECL.

[00377] In the presence of CFA0330-F760G4, CFA0341-F760G4, or human C5 only in the plasma sample, the following method was used. Anti-human C5 antibody (Santa Cruz) was distributed in a bare 96-well MULTI-ARRAY plate (Meso Scale Discovery) and left to stand overnight at 4°C to prepare plates immobilized with anti-human C5 antibody. Calibration curve samples and mouse plasma samples diluted 100 times or more with 1 μg / ml of injected antibody (CFA0330-F760G4 or CFA0341-F760G4) were prepared and incubated for 30 minutes at 37°C. Subsequently, the samples were distributed in the plates immobilized with anti-human C5 antibody and left to stand for one hour at room temperature. Then, anti-human IgG antibody labeled with SULFO-TAG (Meso Scale Discovery) was added and allowed to react for one hour at room temperature, followed by washing.Immediately afterwards, Read Buffer T (x4) (Meso Scale Discovery) was distributed and the measurement was performed using a Sector Imager 2400 (Meso Scale Discovery).

[00378] In the presence of CFA0305-F760G4, CFA0307-F760G4, CFA0366-F760G4, CFA0501-F760G4, CFA0538-F760G4, CFA0599F760G4, CFA0666-F760G4, CFA0672-F760G4 or CFA0675-F760G4 in the plasma sample, the following method was used. Anti-human C5 antibody (CFA0329-F939G4; VH, SEQ ID NO: 23 and VL, SEQ ID NO: 27) was distributed in a bare 96-well MULTI-ARRAY plate (Meso Scale Discovery) and left to stand overnight at 4 degrees C to prepare immobilized plates with anti-human C5 antibody. Samples of calibration curves and plasma samples of Petition 870260079322, dated 07 / 08 / 2026, page 158 / 523 150 / 167 mice diluted 100 times or more with acidic solution (pH 5.5) were prepared and incubated for 30 minutes at 37°C. Subsequently, the samples were distributed onto plates immobilized with anti-human C5 antibody and allowed to stand for one hour at room temperature. Then, SULFO-TAG-labeled anti-human C5 antibody (CFA0300-F939G4; VH, SEQ ID NO: 24 and VL, SEQ ID NO: 28) was added to react for one hour at room temperature and washed. Immediately afterwards, Read Buffer T (x4) (Meso Scale Discovery) was dispensed and the measurement was performed using a Sector Imager 2400 (Meso Scale Discovery).

[00379] The concentration of human C5 was calculated based on the response of the calibration curve using the analytical software SOFTmax PRO (Molecular Devices). The course of C5 concentration in human plasma after intravenous administration, as measured by this method, is shown in Figure 13.The data are represented graphically as the remaining percentage compared to the human plasma concentration of C5 at 5 minutes. 6.3. Measurement of plasma concentration of human anti-C5 antibodies by ECL assay

[00380] The plasma concentration of mouse anti-C5 antibody was measured by ECL. The anti-human IgG antibody fragment (specific for the gamma chain) F(ab')2 (Sigma) or anti-human IgG kappa chain (Antibody Solutions) was distributed in a 96-well MULTI-ARRAY plate (Meso Scale Discovery) and left to stand overnight at 4°C to prepare plates immobilized with anti-human IgG antibody. Calibration curve samples and mouse plasma samples diluted 100 times or more were prepared. Subsequently, the samples were distributed in the plates with immobilized anti-human IgG antibody and left to stand for one hour at room temperature. In Petition 870260079322, dated 07 / 08 / 2026, p. 159 / 523 151 / 167. Subsequently, biotinylated anti-human IgG antibody (Southernbiotech) or SULFOTAG-labeled anti-Fc IgG antibody (Southernbiotech) was added and reacted for one hour at room temperature, followed by washing. Subsequently, only when using biotinylated anti-human IgG antibody, SULFOTAG-labeled streptavidin (Meso Scale Discovery) was added and reacted for one hour at room temperature, followed by washing. Immediately afterward, Read Buffer T (x4) (Meso Scale Discovery) was dispensed, and the measurement was performed using a Sector Imager 2400 (Meso Scale Discovery). The concentration of human anti-C5 antibody was calculated based on the calibration curve response using the analytical software SOFTmax PRO (Molecular Devices). The time course of the plasma concentration of human anti-C5 antibody after intravenous administration, as measured by this method, is shown in Figure 14.The data are presented as the remaining percentage compared to the plasma concentration of human anti-C5 antibody at 5 minutes. 6.4. pH-dependent effect of anti-human C5 antibody binding after in vivo elimination of human C5.

[00381] pH-dependent anti-human C5 antibodies (CFA0305-F760G4, CFA0307-F760G4, CFA0366-F760G4, CFA0501F760G4, CFA0538-F760G4, CFA0599-F760G4, CFA0666-F760G4, CFA0672-F760G4 and CFA0675-F760G4) and pH-independent anti-human C5 antibodies (CFA0330-F760G4 and CFA0341F760G4) were tested in vivo and the resulting plasma concentration of anti-human C5 antibody and the plasma concentration of human C5 were compared. As shown in Figure 14, antibody exposure was comparable. However, the elimination of human C5 administered simultaneously with pH-dependent anti-human C5 antibodies was accelerated when compared to that Petition 870260079322, dated 07 / 08 / 2026, page 160 / 523 152 / 167 of the pH-dependent anti-human C5 antibodies (Figure 13). Example 7 Optimization of anti-C5 monoclonal antibodies (variants 305)

[00382] Several mutations were introduced into the optimized variable region of the anti-C5 antibody, 305LO5, to further enhance its properties, and the optimized variable regions 305LO15, 305LO16, 305LO18, 305LO19, 305LO20, 305LO22, and 305LO23 were generated. The amino acid sequences of VH and VL of the 305 variants are listed in Tables 7 and 8, respectively. The genes encoding the humanized VH were combined with the modified human IgG1 CH variant SG115 (: SEQ ID NO: 114) and the modified human IgG4 CH variants SG422 (: SEQ ID NO: 115) or SG429 (: SEQ ID NO: 116). The genes encoding the humanized VL were combined with a humanized CL (SK1, SEQ ID NO: 38). Separately, heavy and light chain genes encoding a humanized anti-C5 antibody, BNJ441 (BNJ441H, SEQ ID NO: 149; BNJ441L, SEQ ID NO: 150), were synthesized and each was cloned into an expression vector.

[00383] Antibodies were expressed in HEK293 cells cotransfected with a combination of heavy and light chain expression vectors and were purified by protein A. Table 7 Amino acid sequences of VH variants 305 Anticorpo VH HVR-H1 HVR-H2 HVR-H3 305L05 SEQ ID NO: 10 SEQ ID NO: 54 SSYYVA SEQ ID NO: 64 A1YT GSG ATY KASW AKG SEQ ID NO: 74 DGGYDYPTHAMHY 305LO15 SEQ ID NO: 106 SEQ ID NO: 117 SSYYMA SEQ SEQ ID NO: 118 AIFTGSGAEYKAEWAKG SEQ ID NO: 121 DAGYDYPTHAMHY 305LO16 SEQ ID NO: 107 SEQ ID NO: 117 SSYYMA SEQ 108 SEQ ID NO: 117 SSYYMA SEQ ID NO: 118 AIFTGSGAEYKAEWAKG SEQ ID NO: 121 DAGYDYPTHAMHY 305LO19 SEQ ID NO: 109 SEQ ID NO: 117 SSYYMA SEQ ID NO: 118 AIFTGSGAEYKAEWAKG SEQ ID NO: 121 DAGYDYPTHAMHY 305L020 SEQ ID NO: 109 SEQ ID NO: 117 SSYYMA SEQ ID NO: 118 AIFTGSGAEYKAEWAKG SEQ ID NO: 121 DAGYDYPTHAMHY 305LO22 SEQ ID NO: 109 SEQ ID NO: 117 SSYYMA SEQ ID NO: 118 AIFTGSGAEYKAEWAKG SEQ ID NO: 121 D AGYDYPTHAMHY 305LO23 SEQ ID NO: 110 SEQ ID NO: 117 SSYYMA SEQ ID NO: 120 GIFTGSGATYKAEWAKG SEQ ID NO: 121 DAGYDYPTHAMHY Petition 870260079322, on 08 / 07 / 2026, page. 161 / 523 153 / 167 Tabela 8 Sequences of amino acids in VL variants 305 Anticorpo VL HVR-L1 HVR-L2 HVR-L3 305L05 SEQ ID NO: 20 SEQ ID NO: 84 QASQNIGSSLA SEQ ID NO: 94 GASKTHS SEQ ID NO: 104 QSTKVGSSYGNH 305LO15 SEQ ID NO: 111 SEQ ID NO: 122 RASQGISSSLA SEQ ID NO: 123 GASETES SEQ ID NO: 125 QNTKVGSSYG NT 305LO16 SEQ ID NO: 111 SEQ ID NO: 122 RASQGISSSLA SEQ ID NO: 123 GASETES SEQ ID NO: 125 QNTKVGSSYG NT 305LO18 SEQ ID NO: 111 SEQ ID NO: 122 RASQGISSSLA SEQ ID NO: 123 GASETES SEQ ID NO: 125 QNTKVGSSYG NT 305LO19 SEQ ID NO: 111 SEQ ID NO: 122 RASQGISSSLA SEQ ID NO: 123 RASQGISSSLA SEQ ID NO: 123 GASETES SEQ ID NO: 125 QNTKVGSSYG NT 305LO22 SEQ ID NO: 113 SEQ ID NO: 122 RASQGISSSLA SEQ ID NO: 124 GASTTQS SEQ ID NO: 125 QNTKVGSSYG NT 305LO23 SEQ ID NO: 113 SEQ ID NO: 122 RASQGISSSLA SEQ ID NO: 124 GASTTQS SEQ ID NO: 125 QNTKVGSSYG NT Example 8 Characterization of anti-C5 antibody binding (variants 305)

[00384] The kinetic parameters of anti-C5 antibodies against recombinant human C5 were evaluated at 37 degrees C using a BIACORE (registered trademark) T200 instrument (GE Healthcare) under three different conditions; (1) both association and dissociation were at a pH of 744, (2) both association and dissociation were at a pH of 5.8, and (3) association was at a pH of 7.4 and dissociation was at a pH of 5.8. ProA / G (Pierce) was immobilized on a CM1 sensor chip using an amine coupling kit (GE Healthcare) according to the configuration recommended by GE Healthcare. The antibodies and analytes for conditions (1) and (3) were diluted in ACES buffer pH 7.4 (20 mM ACES, 150 mM NaCl, 1.2 mM CaCb, 0.05% Tween 20, 0.005% NaNa) and for condition (2) they were diluted in ACES buffer pH 5.8 (20 mM ACES, 150 mM NaCl, 1.2 mM CaCb, 0.05% Tween 20, 0.005% NaN3). Each antibody was captured on the sensor surface by ProA / G.Antibody uptake levels were typically 60-90 resonance units (RU). Then, recombinant human C5 was... Petition 870260079322, dated 07 / 08 / 2026, page 162 / 523 154 / 167 injected at 3 to 27 nM or 13.3 to 120 nM prepared by three-fold dilution followed by dissociation. The surface was regenerated using 25 mM NaOH. The kinetic parameters in conditions (1) and (2) were determined by fitting the sensorgrams with a 1:1 binding model, and the dissociation rate in condition (3) was determined by fitting the sensorgrams with a 1:1 dissociation to the MCK model using the BIACORE (registered trademark) T200 evaluation program, version 2.0 (GE Healthcare). The pH dependence of all antibodies was shown as a ratio of the dissociation rate of conditions (2) and (1).

[00385] The association rate (ka), dissociation rate (kd), binding affinity (KD), and pH dependence are listed in Table 9. All antibodies showed a faster dissociation rate at pH 5.8 than at pH 7.4, and their pH dependence was around 20 times. Table 9 Kinetic parameters of anti-C5 antibody variants under pH conditions of 7.4 and 5.8 ka (1 / Ms) 7,4_7,4 kd t17s)KD (M) ka (1 / Ms) 5,8_5,8 kd (1 / s)KD (M) 7,4_5,8 kd (1 / s) Dependência do pH L015-SG422 1,40E+064,19E-04 3,00E-10 1,34E+058,79E-03 6,57E-08 1,61E-02 21,0 L015-SG115 1,31E+063,54E-04 2,70E-10 9,49E+048,27E-03 8,72E-08 1,67E-02 23,4 L016-SG422 1,28E+064,12E-04 3,21E-1Q 1,09E+058,69E-03 7,95E-08 1,61E-02 21,1 L018-SG422 1,36E+064,26E-04 3,14E-10 1,39E+058,65E-03 6,24E-08 1,69E-02 20,3 L019-SG422 1,37E+064,76E-04 3,46E-10 1,38E+058,30E-03 6,00E-08 1,61E-02 17,4 LO20-SG115 1,44E+064,67E-04 3,24E-10 1,41E+058,18E-03 5,81 E-08 1,61E-02 17,5 LO20-SG422 1,35E+064,70E-04 3,49E-10 1,36E+058,15E-03 5,99E-08 1,55E-02 17,3 L022-SG115 1,46E+063,82E-04 2,62E-10 1,71E+059,30E-03 5,45E-08 1,46E-02 24,3 L023-SG115 1,53E+064,23E-04 2,77E-10 1,33E+058,55E-03 6,41 E-08 1,73E-02 20,2

[00386] The binding affinity of anti-C5 antibodies (BNJ441, eculizumab, and a 305 variant) to recombinant human C5 at pH 7.4 and pH 5.8 was determined at 37°C using a BIACORE (registered trademark) T200 instrument (GE Healthcare) to assess the effect of pH on antigen binding. The polyclo antibody Petition 870260079322, dated 07 / 08 / 2026, page 163 / 523 155 / 167 goat anti-human IgG (Fc) antibody (KPL #01-10-20) was immobilized on a CM4 sensor chip using an amine coupling kit (GE Healthcare) according to the configuration recommended by GE Healthcare. Antibodies and analytes were diluted in ACES buffer pH 7.4 or ACES buffer pH 5.8 containing 20 mM ACES, 150 mM NaCl, 1.2 mM CaCl2, 0.05% Tween 20, 0.005% NaN3. Antibodies were captured on the sensor surface using the anti-Fc method, and antibody capture levels were typically 50–80 resonance units (RU). Recombinant human C5 was prepared by serial dilution three times starting with 27 nM for assay conditions at pH 7.4 or 135 nM for assay conditions at pH 5.8. The surface was regenerated using 20 mM HCl, 0.01% Tween 20. The data were processed and fitted with a 1:1 linkage model using the BiaEvaluation 2.0 program (GE Healthcare).

[00387] The binding affinity (KD) of BNJ441, eculizumab, and variant 305 with recombinant human C5 at pH 7.4 and pH 5.8 is shown in Table 10. Variant 305 showed a (KD at pH 5.8) / (KD at pH 7.4) ratio of nearly 800, 8 times greater than BNJ441, which showed a (KD at pH 5.8) / (KD at pH 7.4) ratio of only 93. Table 10 Antibody KD (M) pH of 7.4 pH of 5.8 KD ratio at a pH of 5.8 / pH of 7.4 variant 305LO5 1.66E-10 1.32E-07 795 Eculizumab 1.42E-10 2.64E-09 19 BNJ441 1.38E-09 1.28E-07 93 Example 9 Inhibitory activity of anti-C5 antibodies (variants 305) on C5 activation 9.1. Inhibition of complement-activated liposome lysis by Petition 870260079322, dated 07 / 08 / 2026, page 164 / 523 156 / 167 MAbs anti-C5

[00388] Anti-C5 MAbs were tested for complement activity inhibition by liposome lysis assay. Thirty µL of normal human serum (6.7%) (Biopredic, SER019) were mixed with 20 µL of diluted MAb in a 96-well plate and incubated on a shaker for 30 min at room temperature. Liposome solution sensitized with antibodies against dinitrophenyl (Autokit CH50, Wako, 995-40801) was transferred to each well and placed on a shaker for 2 min at 37°C. Fifty µL of substrate solution (Autokit CH50) were added to each well and mixed by shaking for 2 min at 37°C. The final mixture was incubated at 37°C for 40 min and then the OD at 340 nm was measured. The liposome lysis percentage was defined as 100 x [(OD MAb - OD serum and background liposome)] / [(OD without MAb OD serum and background liposome)].Figure 15 shows that the anti-C5 MAbs: 305LO15-SG422, 305LO16-SG422, 305LO18-SG422, 305LO19SG422, 305LO20-SG422, and 305LO20-SG115, inhibited liposome lysis. Two antibodies with Fc variants: 305LO15-SG115 and 305LO23-SG429, also inhibited liposome lysis (Figure 16).

[00389] Anti-C5 MAbs were tested for inhibition of recombinant human C5 (SEQ ID NO: 39). Ten µL of C5-deficient human serum (Sigma, C1163) were mixed with 20 µL of diluted MAb and 20 µL of recombinant C5 (0.1 µg / mL) in a 96-well plate and incubated on a shaker for 1 hour at 37°C. Liposomes (Autokit CH50) were transferred to each well and placed on a shaker for 1 hour at 37°C. Fifty µL of substrate solution (Autokit CH50) were added to each well and mixed by shaking for 2 min at 37°C. The final mixture was incubated at 37°C for 180 min and then OD at 340 nm was Petition 870260079322, dated 07 / 08 / 2026, page 165 / 523 157 / 167 measurement. The percentage of liposome lysis was defined as above. Figure 17 shows that the MAbs: 305LO22-SG115, 305LO22SG422, 305LO23-SG115 and 305LO23-SG422, inhibited liposome lysis. 9.2. Inhibition of C5a generation by anti-C5 MAbs

[00390] Anti-C5 MAbs were tested for C5a generation during liposome lysis to confirm that anti-C5 MAbs inhibit C5 cleavage into C5a and C5b. C5a levels in the supernatants of a liposome lysis assay were quantified using a C5a ELISA kit (R&D systems, DY2037). All MAbs inhibited C5a generation in the supernatants in a dose-dependent manner (Figures 18 and 19). 9.3 Measurement of complement activity in cynomolgus monkey plasma

[00391] Anti-C5 MAbs were tested for inhibition of complement activity in cynomolgus monkey plasma. Anti-C5 MAbs were administered to monkeys (20 mg / kg) and plasma samples were collected periodically until day 56. Chicken red blood cells (cRBCs) (Innovative research, IC05-0810) were washed with buffered gelatin / veronal-saline containing 0.5 mM MgCb and 0.15 mM CaCl2 (GVB++) (Boston BioProducts, IBB-300X) and then sensitized with chicken anti-RBC antibody (Rockland 103-4139) at 1 µg / ml for 15 minutes at 4°C. The cells were then washed with GVB++ and suspended in the same buffer at 1 x 10⁸ cells / ml. In a separate 96-well round-bottom microtest plate, monkey plasma was incubated with sensitized cRBCs at 37°C for 20 minutes. After incubation, the plate was centrifuged at 1000 xg for 2 minutes at 4°C.The supernatants were transferred to the wells of 96-well flat-bottom microtest plates for measurement. Petition 870260079322, dated 07 / 08 / 2026, p. 166 / 523 158 / 167 of OD at 415 nm with a reference wavelength of 630 nm. The percentage of hemolysis was defined as 100 x [(Post-administration OD - Plasma OD and background cRBCs)] / [(Pre-administration OD - Plasma OD and background cRBCs)]. Figure 20 shows that the anti-C5 MAbs: 305LO15-SG422, 305LO15-SG115, 305LO16-SG422, 305LO18-SG422, 305LO19SG422, 305LO20-SG422, 305LO20-SG115 and 305LO23-SG115, inhibited complement activity in plasma. 9.4. Inhibition of the biological activity of C5 variants by anti-C5 MAbs

[00392] Anti-C5 MAbs were tested for inhibition of recombinant human C5 variants: V145I, R449G, V802I, R885H, R928Q, D966Y, S1310N, and E1437D. It has been reported that PNH patients who have the R885H mutation in C5 show a poor response to eculizumab (see, for example, Nishimura et al., New Engl. J. Med. 370: 632-639 (2014)). Each of the human C5 variants was expressed in FS293 cells and the supernatants were used for the following study. Ten microliters of C5-deficient human serum (Sigma, C1163) were mixed with 20 microliters of diluted Mab and 20 microliters of cell culture medium containing a recombinant C5 variant (2-3 micrograms / mL) in a 96-well plate and incubated on a shaker for 0.5 hours at 37°C. Liposomes (Autokit CH50) were transferred to each well and placed on a shaker for 2 minutes at 37°C.Fifty microliters of substrate solution (Autokit CH50) were added to each well and mixed by shaking for 2 min at 37°C. The final mixture was incubated at 37°C for 90 minutes, and then the OD at 340 nm was measured. The percentage of liposome lysis was defined as above. Figure 21 shows that the anti-C5 MAb (eculizumab) did not inhibit the R885H variant of C5, but it did inhibit the other variants tested. Figure 22 shows that the anti-C5 MAb (a 305 variant) inhibited all of them. Petition 870260079322, dated 07 / 08 / 2026, page 167 / 523 159 / 167 variants of C5 tested. 9.5. Inhibition of complement-activated liposome lysis by anti-C5 MAbs

[00393] Anti-C5 MAbs were tested for inhibition of complement activity by liposome lysis assay. Thirty µL of normal human serum (6.7% (Biopredic, SER109)) were mixed with 20 µL of diluted MAb in a 96-well plate and incubated on a shaker for 30 min at room temperature. Liposome solution sensitized with antibodies against dinitrophenyl (Autokit CH50, Wako, 995-40801) was transferred to each well and placed on a shaker for 2 min at 25°C. Fifty µL of substrate solution (Autokit CH50) were added to each well and mixed by shaking for 2 min at 25°C. The final mixture was incubated at 37°C for 45 min, and then the OD at 340 nm was measured. The percentage of liposome lysis inhibition was defined as 100 x [(OD MAb - OD serum and background liposome)] / [(OD without MAb - OD serum and background liposome).Figure 23 shows that the anti-C5 MAbs, BNJ441 and variant 305 inhibited liposome lysis, and that variant 305 has stronger inhibitory activity than BNJ441. Example 10 Pharmacokinetic study of anti-C5 monoclonal antibodies (305 variants) in cynomolgus monkeys 10.1. In vivo trial using cynomolgus monkeys

[00394] The in vivo kinetics of human anti-C5 antibody were evaluated after administration of human anti-C5 antibody to cynomolgus monkeys (Shin Nippon Biomedical Laboratories, Ltd., Japan). A solution of human anti-C5 antibody (2.5 mg / ml) was administered once at a dose of approximately 8 ml / kg into the cephalic vein of the forearm during a 30-minute infusion. Blood was collected pre-administration and at 5 minutes, seven hours, one day, two days, three Petition 870260079322, dated 07 / 08 / 2026, page 168 / 523 160 / 167 days, seven days, fourteen days, twenty-one days, twenty-eight days, thirty-five days, forty-two days, forty-nine days, and fifty-six days after administration. The collected blood was immediately centrifuged at 1,700 x e and 4 degrees C for 10 minutes to separate the plasma. The separated plasma was stored in a freezer at -70 degrees C or below before assay. Anti-human C5 antibodies were prepared as described in Example 7. 10.2. Measurement of total plasma C5 concentration in cynomolgus monkeys by an ELISA assay.

[00395] The total concentration of C5 in cynomolgus monkey plasma was measured by ELISA. Human anti-C5 antibody (internal antibody generated using the method described in Example 2) was distributed onto Nunc-ImmunoPlate MaxiSorp plates (Nalge Nunc International) and left to stand overnight at 4 °C to prepare plates immobilized with cynomolgus monkey anti-C5 antibody. Calibration curve samples and cynomolgus monkey plasma samples diluted 20,000 times with 0.4 µg / ml of injected antibody were prepared and incubated for 60 minutes at 37 °C. Subsequently, the samples were distributed onto plates with immobilized cynomolgus monkey anti-C5 antibody and left to stand for one hour at room temperature. Next, HRP-labeled anti-human IgG antibody (SouthernBiotech) was added and allowed to react for thirty minutes at room temperature, followed by washing.Subsequently, the ABTS ELISA HRP (KPL) substrate was added. The signal was measured by a plate reader at a wavelength of 405 nm. The concentration of cynomolgus monkey C5 was calculated based on the calibration curve response using the analytical software SOFTmax PRO (Molecular Devices). The time course of the concentration of cynomolgus monkey C5 in plasma after intravenous administration, as measured by... Petition 870260079322, dated 07 / 08 / 2026, page 169 / 523 161 / 167 this method, is shown in Figure 24. The data are represented as the remaining percentage compared to the concentration of cynomolgus monkey C5 in plasma at pre-administration. The pH-dependent anti-human C5 antibodies (305LO15-SG422, 305LO15SG115, 305LO16-SG422, 305LO18-SG422, 305LO19-SG422, 305LO20-SG422, 305LO20-SG115, 305LO22-SG422, 305LO23SG422 and 305LO23-SG115) showed less plasma accumulation of C5 when compared with pH-dependent anti-human C5 antibodies. 10.3. Measurement of plasma concentration of human anti-C5 antibody by ELISA assay

[00396] The concentration of human anti-C5 antibody in cynomolgus monkey plasma was measured by ELISA. Human IgG kappa chain anti-antibody (Antibody Solutions) was distributed onto Nunc-ImmunoPlate MaxiSorp plates (Nalge Nunc International) and left to stand overnight at 4°C to prepare immobilized plates with human anti-IgG. Calibration curve samples and cynomolgus monkey plasma samples diluted 100 times or more were prepared. Subsequently, the samples were distributed onto plates immobilized with human anti-IgG antibody and left to stand for one hour at room temperature. Then, HRP-labeled human anti-IgG antibody (SouthernBiotech) was added and reacted for thirty minutes at room temperature, followed by washing. Subsequently, ABTS ELISA HRP substrate (KPL) was added. The signal was measured by a plate reader at a wavelength of 405 nm.The concentration of human anti-C5 antibody was calculated based on the response of the calibration curve using the analytical software SOFTmax PRO (Molecular Devices). The time course of the plasma concentration of human anti-C5 antibody after intravenous administration, as measured by this software. Petition 870260079322, dated 07 / 08 / 2026, page 170 / 523 The pH-dependent anti-human C5 antibodies (305LO15-SG422, 305LO15-SG115, 305LO16SG422, 305LO18-SG422, 305LO19-SG422, 305LO20-SG422, 162 / 167 method) are presented in Figure 25. 305LO20-SG115, 305LO22-SG422, 305LO23-SG422 and 305LO23SG115) exhibited a longer half-life when compared to pH-independent anti-human C5 antibodies. Example 11 X-ray crystal structure analysis of a Fab complex of a variant 305 and the MG1 domain of human C5. 11.1. Expression and purification of the MG1 (20-104) domain of human C5.

[00397] The MG1 domain (amino acid residues 20 1 104 of : SEQ ID NO: 29) fused to a GST marker via a cleavable thrombin ligand (GST-MG1) was expressed in the E. coli strain BL21 DE3 pLysS (promega) using a pGEX-4T-1 vector (GE Healthcare). Protein expression was induced with 0.1 mM isopropyl betaD-1-thiogalactopyranoside (IPTG) at 25°C for 5 hours. The bacterial cell pellet was lysed with Bugbuster (Merck) supplemented with lysonase (Merck) and a complete protease inhibitor cocktail (Roche), followed by purification of GST-MG1 from the soluble fraction using the GSTrap column (GE Healthcare) according to the manufacturer's instructions. The GST marker was cleaved with thrombin (Sigma) and the resulting MG1 domain was further purified using a Superdex 75 gel filtration column (GE Healthcare). The fractions containing the MG1 domain were pooled and stored at -80 degrees C. 11.2. Preparation of the Fab fragment of a variant 305

[00398] Fab fragments of one of the optimized variants of 305 were prepared by the conventional method using limited digestion with papain (Roche Diagnostics, Cat. No. 1047825), followed Petition 870260079322, dated 07 / 08 / 2026, page 171 / 523 163 / 167 by loading onto a protein A column (MabSlect SuRe, GE Healthcare) to remove the Fc fragments, a cation exchange column (HiTrap SP HP, GE Healthcare), and a gel filtration column (Superdex200 16 / 60, GE Healthcare). The fractions containing the Fab fragment were pooled and stored at -80 degrees C. 11.3. Preparation of a complex with Fab variant 305 and the MG1 domain of human C5

[00399] The purified recombinant human C5 MG1 domain was mixed with a Fab fragment from a 305 variant in a 1:1 molar ratio. The complex was purified by gel filtration chromatography (Superdex200, 10 / 300 magnification, GE Healthcare) using a balanced column with 25 mM HEPES pH 7.5, 100 mM NaCl. 11.4. Crystallization

[00400] The purified complexes were concentrated to approximately 10 mg / mL and crystallization was performed by the suspended drop vapor diffusion method in combination with the seeding method at 4°C. The reservoir solution consisted of 0.2 M dehydrated magnesium formate, 15% w / v polyethylene glycol 3350. This resulted in the yield of plate-like crystals in a few days. The crystal was immersed in a solution of 0.2 M dehydrated magnesium formate, 25% w / v polyethylene glycol 3350, and 20% glycerol. 11.5. Data collection and structure determination

[00401] X-ray diffraction data were measured by BL32XU in Spring-8. During the measurement, the crystal was constantly placed in a nitrogen flow at -178 degrees C to maintain the frozen state and a total of 180 X-ray diffraction images were collected using an MX-225HS CCD detector (RAYONIX) coupled to a beamline, while the crystal was rotated 1.0 degree at a time. Petition 870260079322, dated 07 / 08 / 2026, page 172 / 523 164 / 167 The determination of cellular parameters, indexing of diffraction points, and processing of diffraction data obtained from diffraction images were performed using the Xia2 program (J. Appl. Cryst. 43: 186-190 (2010), XDS Package (Acta. Cryst. D66: 125132 (2010)), and Scala (Acta. Cryst. D62: 72-82 (2006)). Finally, diffraction intensity data up to a resolution of 2.11 Angstroms were obtained. Statistical crystallographic data are shown in Table 11. Table 11 I STOPPED Collection of X-ray data and statistical refinements Data Collection Spatial Group P1 Cellular Unit a,b,c (A) α,β,γ(°) 39.79, 55.10, 127.76 89.18 ,86.24, 78.20 Resolution (A) 49.19 -2.11 Total Reflections 112.102 Single Reflections 56.154 Integrity (highest resolution layer) (%) 92.1 (95.8) Rmergea (highest resolution layer) (%) 7.2 (31.7) Refinement Resolution (A) 25.00 -2.11 Reflections 53.398 Rb Factor (Rfreec) (%) 20.42 (26.44) rms deviation from ideal Bond Lengths (A) Bond Angles (°) 0.0088 1.3441 a: imerge = Ihkllj / ij (hkl) - ( / (hkl)} | IlhkíLjhj (hkl), where ij (hkl) and ( / (hkl)} are the intensity of the measurement and the average intensity for the reflection with the indices hkl, respectively. b; factor R = lhkl\FCa\c (hkí)\ - |FObs (hkí)\l lhkl\FObs (hkl), where Fobs and FCa\c are the amplitudes of the observed and calculated structure factor, respectively. c; Rhee is calculated with 5% of the reflection randomly left aside.

[00402] The structure was determined by molecular substitution using the Phaser program (J. Appl. Cryst. 40: 658-674 (2007)). The Fab domain research model was derived from the published crystal structure of human lgG4 Fab (PDB code: 1BBJ) and the MG1 domain research model was from the published crystal structure of human C5 (PDB code: 3CU7, Nat. Immunol. 9: 753-760 (2008)). A model was constructed using the Coot program (Acta Cryst. D66: 486-501 (2010)) and refined using the Refmacõ program (Acta Cryst. D67: 355367 (2011)). The crystallographic reliability factor (R) for the data Petition 870260079322, dated 07 / 08 / 2026, page 173 / 523 The diffraction intensity of 165 / 167 at 25-2.11 Angstroms was 20.42%, with a Free R value of 26.44%. The structure refinement statistics are shown in Table 11. 11.6. Global structure of a Fab complex of variant 305 and the MG1 domain of C5

[00403] The Fab fragment of a 305 variant (305 Fab) linked to the MG1 domain of C5 (MG1) in a 1:1 ratio and the asymmetric unit of the crystal structure contained in the two complexes, Molecules 1 and 2, as illustrated in Figure 26A. Molecules 1 and 2 can be well aligned in an RMSD of 0.717 Angstrom with the C-alpha atom position in all residues, as shown in Figure 26B. The figures discussed below were prepared using Molecule 1.

[00404] In Figures 27A and 27B, the contact region epitope of Fab 305 is mapped onto the amino acid sequence of MG1 and the crystal structure, respectively. The epitope includes MG1 amino acid residues containing one or more atoms located within 4.5 Angstroms of any part of Fab 305 in the crystal structure. Additionally, the epitope at 3 Angstrom(s) is highlighted in Figure 27A. 11.7. Interactions of E48, D51 and K109

[00405] As described in Examples 4.5 and 4.6, anti-C5 MAbs including antibodies of the 305 series were tested for binding to human C5 point mutants E48A, D51A, and K109A by blot binding and BIACORE (trademark) analyses. Although the 305 variants bind strongly to WT C5, they bind only weakly to the E48A C5 mutant and do not bind to the D51A and K109A mutants. The crystal structure of the 305 and MG1 Fab complex revealed that the three amino acids E48, D51, and K109 are all 3.0 Angstroms away from the 305 Fab, forming multiple linkages. Petition 870260079322, dated 07 / 08 / 2026, p. 174 / 523 166 / 167 hydrogen bonds with Fab, as shown in Figure 28A. Upon closer examination, the K109 prison of MG1 is embedded in a notch formed at the Fab heavy chain interface and interacts intimately through three hydrogen bonds with H-CDR3_G97, HCDR3_Y100, and H-CDR3_T100b, and through a salt bond with HCDR3_D95 (Figure 28D). D51 is located between MG1 and the Fab heavy chain of 305 and makes two hydrogen bonds with HCDR1_Ser32 and H-CDR2_Ser54 to fill the gap (Figure 28C). This indicates that K109 and D51 of C5 are both critical residues for antibody binding in the 305 series. On the other hand, E48 is located closer to the surface and forms only one hydrogen bond with Fab, suggesting that its contribution to antibody binding may be less than that of K109 and E51 (Figure 28B). These relationships are consistent with the results of Western blot and BIACORE (trademark) binding analyses of human C5 mutants (Examples 4.5 and 4.6).6) Additional note: The numbering of Fab amino acid residues is based on the Kabat numbering scheme. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991). 11.8. Interactions of H70, H72, and H110 of human C5 and the 305 series of antibodies.

[00406] Crystalline structure analysis revealed that the three histidine residues on human C5, namely H70, H72, and H110, are included in the Fab epitope of variant 305, as shown in Figure 27A and Figure 29A. A BIACORE (registered trademark) linkage analysis was performed to investigate the contribution of these histidine residues to the pH-dependent protein-protein interaction between human C5 and Fab variant 305, using the human C5 mutants H70Y, H72Y, H110Y, and H70Y+H110Y (Example 4.7). H72Y Petition 870260079322, dated 07 / 08 / 2026, page 175 / 523 The 167 / 167 mutation resulted in the complete loss of the Fab variant 305's attachment to C5. This C5 residue is located in a pocket formed by the CDR2 loop of the Fab 305 heavy chain and the MG1 loop (L73, S74, and E76) and tightly fills this space, as shown in Figure 29C. Furthermore, the H72 residue of C5 forms a hydrogen bond with H-CDR2_Y58. The H72Y mutation should not be expected to be tolerated as there is insufficient space to accommodate the bulky tyrosine side chain. A hydrogen bond with H-CDR2_Y58 cannot be maintained. Regarding the contribution of H70 and H110 to pH dependence, the H70Y and H110Y mutations resulted in slower dissociation of the Fab varia...

Claims

1. Use of an antibody that binds to C5, characterized in that it is in the manufacture of a medicament for the treatment of a complement-mediated disease or condition involving excessive or uncontrolled activation of C5, wherein the antibody binds to an epitope within the beta chain of C5 with a higher affinity at neutral pH than at acidic pH, wherein the antibody comprises an Fc region in which lysine at residue 447 according to EU numbering is not present.

2. Use of an antibody that binds to C5, characterized by the fact that it is in the manufacture of a drug to increase the removal of C5 from plasma, wherein the antibody binds to an epitope within the beta chain of C5 with a higher affinity at neutral pH than at acidic pH, wherein the antibody comprises an Fc region in which lysine at residue 447 according to EU numbering is not present.

3. Use, according to any one of claims 1 to 2, characterized in that the antibody has a feature selected from the group consisting of: (a) the antibody contacts amino acids D51 and K109 of C5 (SEQ ID NO: 39); (b) the antibody's affinity for C5 (SEQ ID NO: 39) is greater than the antibody's affinity for a C5 mutant consisting of an E48A substitution of SEQ ID NO: 39; and (c) the antibody binds to a C5 protein consisting of the amino acid sequence of SEQ ID NO: 40 at a pH of 7.4, but does not bind to a C5 protein consisting of the amino acid sequence of SEQ ID NO: 39 with an H72Y substitution at a pH of 7.

4.

4. Use, according to any of claims 1 to Petition 870260079322, dated 07 / 08 / 2026, page 177 / 523 2 / 8 3, characterized in that the antibody competes for binding to C5 with an antibody comprising a VH and VL pair selected from: (a) a VH of SEQ ID NO: 1 and a VL of SEQ ID NO: 11; (b) a VH of SEQ ID NO: 5 and a VL of SEQ ID NO: 15; (c) a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 14; (d) a VH of SEQ ID NO: 6 and a VL of SEQ ID NO: 16; (e) a VH of SEQ ID NO: 2 and a VL of SEQ ID NO: 12; (f) a VH with SEQ ID NO: 3 and a VL with SEQ ID NO: 13; (g) a VH with SEQ ID NO: 9 and a VL with SEQ ID NO: 19; (h) a VH with SEQ ID NO: 7 and a VL with SEQ ID NO: 17; (i) a VH with SEQ ID NO: 8 and a VL with SEQ ID NO: 18; and (j) a VH with SEQ ID NO: 10 and a VL with SEQ ID NO:

20.

5. Use, according to any one of claims 1 to 4, characterized in that the antibody binds to: (a) an epitope within the MG1-MG2 domain of the beta chain of C5; (b) an epitope within a fragment consisting of amino acids 33-124 of the beta chain (SEQ ID NO: 40) of C5; (c) an epitope within the beta chain (SEQ ID NO: 40) of Petition 870260079322, dated 07 / 08 / 2026, p. 178 / 523 3 / 8 C5 comprising at least one fragment selected from the group consisting of amino acids 47-57, 70-76 and 107-110; (d) an epitope within a beta-chain fragment (SEQ ID NO: 40) of C5 comprising at least one amino acid selected from the group consisting of Glu48, Asp51, His70, His72, Lys109, and His110; and / or (e) the same epitope as an antibody described in Tables 2, 7, or 8.

6. Use, according to any one of claims 1 to 5, characterized in that the antibody inhibits the activation of C5.

7. Use, according to any one of claims 1 to 6, characterized in that the antibody inhibits the activation of the C5 R885H variant.

8. Use, according to any one of claims 1 to 7, characterized in that the antibody is a monoclonal antibody.

9. Use, according to any one of claims 1 to 8, characterized in that the antibody is a human, humanized or chimeric antibody.

10. Use, according to any one of claims 1 to 9, characterized in that the antibody is an antibody fragment that binds to C5.

11. Use, according to any one of claims 1 to 10, characterized in that the antibody comprises (a) an HVR-H3 comprising the amino acid sequence DX1GYX2X3PTHAMX4X5, wherein X1 is G or A, X2 is V, Q or D, X3 is T or Y, X4 is Y or H, X5 is L or Y (SEQ ID NO: 128), (b) an HVR-L3 comprising the amino acid sequence QX1TX2VGSSYGNX3, wherein Xi is S, C, N or T, X2 is F or K, X3 is A, T or H (SEQ ID NO: 131) and (c) an HVR-H2 comprising the amino acid sequence Petition 870260079322, dated 07 / 08 / 2026, page. 179 / 523 4 / 8 X1IX2TGSGAX3YX4AX5WX6KG, where Xi is C, A or G, X2 is Y or F, X3 is T, D or E, X4 is Y, K or Q, X5 is S, D or E, X6 is A or V (SEQ ID NO: 127).

12. Use, according to any one of claims 1 to 11, characterized in that the antibody comprises (a) an HVR-H1 comprising the amino acid sequence SSYYX1X2, wherein X1 is M or V, X2 is C or A (SEQ ID NO: 126), (b) an HVR-H2 comprising the amino acid sequence X1IX2TGSGAX3YX4AX5WX6KG, wherein X1 is C, A or G, X2 is Y or F, X3 is T, D or E, X4 is Y, K or Q, X5 is S, D or E, X6 is A or V (SEQ ID NO: 127) and (c) an HVR-H3 comprising the amino acid sequence DX1GYX2X3PTHAMX4X5, wherein X1 is G or A, X2 is V, Q or D, X3 is T or Y, X4 is Y or H, X5 is L or Y (SEQ ID NO: 128).

13. Use according to claim 12, characterized in that the antibody further comprises (a) an HVR-L1 comprising the amino acid sequence X1ASQX2IX3SX4LA, wherein X1 is Q or R, X2 is N, Q or G, X3 is G or S, X4 is D, K or S (SEQ ID NO: 129); (b) an HVR-L2 comprising the amino acid sequence GASX1X2X3S, wherein X1 is K, E or T, X2 is L or T, X3 is A, H, E or Q (SEQ ID NO: 130); and (c) an HVR-L3 comprising the amino acid sequence QX1TX2VGSSYGNX3, wherein X1 is S, C, N or T, X2 is F or K, X3 is A, T or H (SEQ ID NO: 131).

14. Use, according to any one of claims 1 to 10, characterized in that the antibody comprises (a) an HVR-L1 comprising the amino acid sequence X1ASQX2IX3SX4LA, wherein X1 is Q or R, X2 is N, Q or G, X3 is G or S, X4 is D, K or S (SEQ ID NO: 129); (b) an HVR-L2 comprising the amino acid sequence GASX1X2X3S, wherein X1 is K, E or T, X2 is L or T, X3 is A, H, E or Q (SEQ ID NO: 130); and (c) an HVR-L3 comprising the amino acid sequence QX1TX2VGSSYGNX3, in Petition 870260079322, dated 07 / 08 / 2026, p. 180 / 523 5 / 8 where X1 is S, C, N, or T, X2 is F or K, X3 is A, T, or H (SEQ ID NO: 131).

15. Use, according to any one of claims 1 to 10, characterized in that the antibody comprises: (1) a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118 and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 123; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125; (2) a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 119 and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122;(b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 123; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125; (3) a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118 and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 124; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125;or Petition 870260079322, of 07 / 08 / 2026, p. 181 / 523 6 / 8 (4) a VH sequence containing (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 117, (b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 120 and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 121 and a VL sequence containing (a) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 122; (b) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 124; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 125.; 16. Use, according to claim 12, characterized in that the antibody further comprises a variable heavy chain domain structural region FR1 comprising the amino acid sequence of any of SEQ ID NOs: 132-134; a variable heavy chain domain structural region FR2 comprising the amino acid sequence of any of SEQ ID NOs: 135-136; a variable heavy chain domain structural region FR3 comprising the amino acid sequence of any of SEQ ID NOs: 137-139; and a variable heavy chain domain structural region FR4 comprising the amino acid sequence of any of SEQ ID NOs: 140-141.

17. Use, according to claim 14 or 15, characterized in that the antibody further comprises a variable light chain domain structural region FR1 comprising the amino acid sequence of any of SEQ ID NOs: 142-143; a variable light chain domain structural region FR2 comprising the amino acid sequence of any of SEQ ID NOs: 144-145; a variable light chain domain structural region FR3 comprising the amino acid sequence of any of SEQ ID NOs: 146-147; and a variable light chain domain structural region FR4 comprising the amino acid sequence of SEQ ID NO:

148.

18. Use, according to any one of claims 1 to 10, characterized in that the antibody comprises (a) a VH sequence having at least 95% sequence identity with the amino acid sequence of any one of SEQ ID NOs: 10, 106-110; (b) a VL sequence having at least 95% sequence identity with the amino acid sequence of any one of SEQ ID NOs: 20, 111-113; or (c) a VH sequence conforming to (a) and a VL sequence conforming to (b).

19. Use according to claim 18, characterized in that the antibody comprises a VH sequence of any one of SEQ ID NOs: 10, 106-110, and / or the antibody comprises a VL sequence of any one of SEQ ID NOs: 20, 111-113.

20. Use, according to claim 19, characterized in that the antibody comprises a VH sequence of any one of SEQ ID NOs: 10, 106-110 and a VL sequence of any one of SEQ ID NOs: 20, 111-113.

21. Use, according to any one of claims 1 to 10, characterized in that the antibody comprises: (a) a VH sequence of SEQ ID NO: 106 and a VL sequence of SEQ ID NO: 111; (b) a VH sequence of SEQ ID NO: 107 and a VL sequence of SEQ ID NO: 111; (c) a VH sequence of SEQ ID NO: 108 and a VL sequence of SEQ ID NO: 111; (d) a VH sequence of SEQ ID NO: 109 and a VL sequence of SEQ ID NO: 111; (e) a VH sequence of SEQ ID NO: 109 and a VL sequence of SEQ ID NO: 112; Petition 870260079322, dated 07 / 08 / 2026, p. 183 / 523 8 / 8 (f) a VH sequence from SEQ ID NO: 109 and a VL sequence from SEQ ID NO: 113; or (g) a VH sequence from SEQ ID NO: 110 and a VL sequence from SEQ ID NO:

113.

22. Use, according to any one of claims 1 to 20, characterized in that the antibody is a full-length IgG1 or IgG4 antibody.

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