Monoclonal antibodies, compositions and methods for detecting complement factor d

Abstract

To provide monoclonal antibodies and compositions comprising such antibodies for use in detecting the presence and amount of mature Factor D and Pro-Factor D.SOLUTION: Disclosed herein are monoclonal antibodies that specifically bind to human mature Factor D and that do not bind to human Pro-Factor D, monoclonal antibodies that specifically bind to human Pro-Factor D and do not bind to human mature Factor D, and monoclonal antibodies that bind to both human mature Factor D and human Pro-Factor D. Also disclosed are methods of using the monoclonal antibodies, and compositions comprising the same, to detect the mature and / or the pro-form of Factor D in biological samples, to determine the status of the Alternative Pathway of Complement (APC) in a mammalian subject, or to determine the status of Factor D after treatment with a MASP-3 inhibitory agent which inhibits the conversion of Pro-Factor D to mature Factor D.SELECTED DRAWING: Figure 18

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 066,942, filed August 18, 2020, U.S. Provisional Application No. 63 / 066,948, filed August 18, 2020, and U.S. Provisional Application No. 63 / 197,833, filed June 7, 2021, which are incorporated by reference in their entireties.

[0002] FIELD OF THE INVENTION The present invention relates to monoclonal antibodies for use in detecting the presence and amount of mature factor D and pro-factor D, and compositions comprising such antibodies.

[0003] Sequence Listing Statement The Sequence Listing for this application is provided in text format in lieu of a paper copy and is incorporated herein by reference. The name of the text file containing the Sequence Listing is: MP_1_0316_PCT_Sequence_Listing_20210816_ST25. This text file is 139 KB; it was created on August 16, 2021, and has been submitted via EFS-Web along with the submission of this application. [Background technology]

[0004] background The complement system supports innate host defense against pathogens, and dysregulated and unabated complement activity can also function as a major driver of autoimmune disease, causing uncontrolled inflammation and the spread of tissue destruction. However, dysregulated and unabated complement activity can also function as a major driver of disease, causing uncontrolled inflammation and the spread of tissue destruction. The alternative complement pathway (APC) is generally described as a downstream amplifier of complement activity, enhancing the host immune response following complement activation via the classical and lectin pathways. However, the APC's ability to generate a positive feedback loop of protease complexes that actively drive the formation of new complexes of the same type is unique within the complement pathway (Lachmann PJ, Adv Immunol 104:115-49, 2009).

[0005] Complement factor D (CFD) is a serine protease essential for APC activation. Factor D cleaves factor B bound to C3b to generate the C3b / Bb enzyme, the active component of the alternative pathway C3 / C5 convertase. While CFD is expressed as an inactive zymogen (referred to herein as "pro-factor D"), CFD circulates in plasma primarily as a cleaved mature serine protease (referred to herein as "mature factor D"). As described in WO2013 / 180834 (Patent Document 1) and WO2013 / 192240 (Patent Document 2), it has recently been determined that MASP-3 is responsible for converting complement factor D (CFD) from its zymogen form (pro-factor D) to its activated form (mature factor D), and thus the MASP-3 protein plays an important upstream regulatory role in APCs. As further described in WO2018 / 026722 (Patent Document 3), which is incorporated herein by reference, a number of high-affinity anti-MASP-3 inhibitory antibodies have been produced that bind to the serine protease domain of MASP-3 and inhibit its catalytic activity.

[0006] A current problem in the field of complement research is that anti-factor D antibodies in commercially available test kits do not distinguish between pro-factor D and the activated form (mature factor D). In the plasma of wild-type animals or humans, the majority of systemic factor D has already been processed to its mature form by in vivo MASP-3 activity, making in vitro assessment of APC inhibition by MASP-3 inhibitors impossible using conventional assays. Therefore, there is a need for detection reagents and assays for measuring the presence and amount of pro-factor D and / or mature factor D in biological samples for use as biomarkers of APC status, thereby enabling in vitro assessment of APC inhibition by MASP-3 inhibitors. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] WO2013 / 180834 [Patent Document 2] WO2013 / 192240 [Patent Document 3] WO2018 / 026722 [Non-patent literature]

[0008] [Non-Patent Document 1] Lachmann PJ, Adv Immunol 104:115-49, 2009 Summary of the Invention

[0009] overview This Summary is provided to introduce various concepts in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.

[0010] The present invention addresses the need for detection reagents and assays for determining the presence and amount of pro-factor D and / or mature factor D in biological samples.

[0011] In one aspect, the present disclosure provides an isolated antibody, or antigen-binding fragment thereof, that specifically binds to an epitope in the amino-terminal region of human maturation factor D, the epitope comprising or consisting of the amino acid sequence ILGGREA (SEQ ID NO:5). In one embodiment, the isolated antibody or fragment thereof specifically binds to human maturation factor D (SEQ ID NO:3) and does not bind to human pro-factor D (SEQ ID NO:2). In one embodiment, the antibody is a monoclonal antibody. In one embodiment, the present disclosure provides a nucleic acid molecule encoding the heavy chain variable region CDRs and / or the light chain variable region CDRs of an antibody or fragment thereof that specifically binds to human maturation factor D.

[0012] In another aspect, the present disclosure provides an isolated antibody, or antigen-binding fragment thereof, that specifically binds to an epitope on the activation ("pro") peptide of human factor D, the epitope comprising or consisting of "APPRGR" (SEQ ID NO:4). In one embodiment, the antibody specifically binds to human pro-factor D (SEQ ID NO:2) and does not bind to mature factor D (SEQ ID NO:3). In one embodiment, the antibody is a monoclonal antibody. In one embodiment, the present disclosure provides a nucleic acid molecule encoding the heavy chain variable region CDRs and / or the light chain variable region CDRs of an antibody or fragment thereof that specifically binds to human pro-factor D.

[0013] In another aspect, the present disclosure provides a kit for detecting the presence or amount of maturation factor D and / or pro-factor D in a test sample, the kit comprising: (a) at least one container; and (b) at least one antibody or fragment thereof that specifically binds to human maturation factor D and / or pro-factor D.

[0014] In another aspect, the present disclosure provides an isolated antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human pro-factor D, wherein the antibody comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85-88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89-93, wherein the CDRs are numbered according to the Kabat numbering system.

[0015] In another aspect, the present disclosure provides a method for determining the presence or amount of maturation factor D in a test sample, comprising: (a) contacting the test sample with a maturation factor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or amount of the antibody or fragment thereof bound to maturation factor D, wherein the presence of binding indicates the presence or amount of maturation factor D in the sample; and wherein the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof binds to an epitope in the N-terminal region of maturation factor D represented by amino acid ILGGREA (SEQ ID NO:5).

[0016] In another aspect, the present disclosure provides a method for determining the presence or amount of pro-factor D in a test sample, comprising: (a) contacting the test sample with an anti-human pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or amount of the antibody or fragment thereof bound to pro-factor D, wherein the presence of binding indicates the presence or amount of pro-factor D in the sample; and wherein the anti-human mature pro-factor D-specific antibody or antigen-binding fragment thereof specifically binds to an epitope in the activation ("pro") peptide of human factor D designated as "APPRGR" (SEQ ID NO:4).

[0017] In another aspect, the disclosure provides a method for assessing the degree of alternative pathway complement (APC) activation in a test sample, comprising: (a) providing the test sample; (b) performing an immunoassay comprising at least one of: (i) capturing and detecting mature factor D in the test sample, wherein the mature factor D is captured or detected with a mature factor D-specific monoclonal antibody or fragment thereof that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but does not bind to pro-factor D; and / or (ii) capturing and detecting pro-factor D in the test sample, wherein the pro-factor D is captured or detected with a pro-factor D-specific monoclonal antibody or fragment thereof that specifically binds to an epitope on the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but does not bind to mature factor D; and (c) (b)(i) comparing the level of mature factor D detected according to (b)(i) with a predetermined level or a control sample, and / or (b)(ii) comparing the level of pro-factor D detected according to (b)(ii) with a predetermined level or a control sample, wherein the level of mature factor D and / or pro-factor D detected in the test sample is indicative of the degree of alternative pathway complement activation.

[0018] In another aspect, the present disclosure provides a method for monitoring the effectiveness of treatment with a MASP-3 inhibitory antibody in a mammalian subject, the method comprising: (a) administering a fixed dose of a MASP-3 inhibitory antibody to the mammalian subject at a first time point; (b) assessing a first concentration of maturation factor D and / or pro-factor D in a biological sample obtained from the subject after step (a); (c) treating the subject with the MASP-3 inhibitory antibody at a second time point; (d) assessing a second concentration of maturation factor D and / or pro-factor D in a biological sample obtained from the subject after step (c); and (e) comparing the level of maturation factor D and / or pro-factor D assessed in step (b) with the level of maturation factor D and / or pro-factor D assessed in step (d) to determine the effectiveness of the MASP-3 inhibitory antibody in the mammalian subject.

[0019] In another aspect, the present disclosure provides a method of treating a mammalian subject suffering from or at risk of developing an alternative pathway disease or disorder, comprising administering a MASP-3 inhibitory antibody to the subject when the subject is determined to have (i) a lower or decreased level of pro-factor D in one or more samples taken from the subject compared to a predetermined pro-factor D level or compared to a pro-factor D level in one or more control samples; and / or (ii) a higher or increased level of mature factor D in one or more samples taken from the subject compared to a predetermined mature factor D level or compared to a mature factor D level in one or more control samples.

[0020] In another aspect, the present disclosure provides a pharmaceutical composition comprising a MASP-3 inhibitory antibody in an aqueous solution comprising a buffer system having pH 6.0±5%, 20±5% mM histidine, 100±5% mg / mL sucrose, and 0.035%±5% polysorbate 80, wherein the MASP-3 inhibitory antibody is present at a concentration of 110 mg / mL±5%, and the MASP-3 inhibitory antibody has an HC-CDR1 comprising SEQ ID NO:231 (GKWIE); TIFF2025076475000002.tif4128 or and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO:239, an LC-CDR2 comprising SEQ ID NO:178 (WASTRES); and an LC-CDR3 comprising SEQ ID NO:244 (KQSYNIPT).

[0021] In another aspect, the present disclosure provides an article of manufacture containing a pharmaceutical composition comprising a MASP-3 inhibitory antibody, wherein the MASP-3 inhibitory antibody is in a unit dosage form of 10 mg to 1000 mg suitable for therapeutic administration to a human subject. [The present invention 1001] An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in the amino-terminal region of human maturation factor D, wherein the epitope comprises or consists of the amino acid sequence ILGGREA (SEQ ID NO:5). [The present invention 1002] 1001. An isolated antibody or antigen-binding fragment thereof of the present invention, which specifically binds to human maturation factor D (SEQ ID NO:3) and does not bind to human pro-factor D (SEQ ID NO:2). [The present invention 1003] 1003. The isolated antibody or antigen-binding fragment thereof of claim 1001 or 1002, wherein said antibody is a monoclonal antibody. [The present invention 1004] The isolated antibody or antigen-binding fragment thereof of any of claims 1001 to 1003, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. [The present invention 1005] An isolated antibody or antigen-binding fragment thereof according to any one of claims 1001 to 1004 of the present invention, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. [The present invention 1006] The isolated antibody or antigen-binding fragment thereof of any one of 1001 to 1005 of the present invention, which is a single-chain molecule. [The present invention 1007] 1007. The isolated antibody or antigen-binding fragment thereof of any of claims 1001 to 1006, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. [The present invention 1008] K<10 nM D 1007. An isolated antibody or antigen-binding fragment thereof of any one of claims 1001 to 1007, which binds to human maturation factor D at 1007. [The present invention 1009] 9. The isolated antibody or antigen-binding fragment thereof of any of claims 1001 to 1008, which is labeled with a detectable moiety. [The present invention 1010] An isolated antibody or antigen-binding fragment thereof according to any one of 1001 to 1009 of the present invention, which is immobilized on a substrate. [The present invention 1011] An isolated antibody or antigen-binding fragment thereof according to any one of claims 1001 to 1010, wherein the isolated antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, and the CDRs are numbered according to the Kabat numbering system. [The present invention 1012] The antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D has the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO: 65), where X at position 1 is T, I, or S, and X at position 3 is G or I; (b) amino acid sequence HC-CDR2 containing TIFF2025076475000004.tif4128 (wherein X at position 11 is H or N and X at position 16 is S or R); (c) amino acid sequence HC-CDR3 containing TIFF2025076475000005.tif4128 (in which X at position 6 is R, G, or N, X at position 7 is S or Y, X at position 8 is F, I, or V, and X at position 10 is D or H); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69) (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54). 10. The isolated antibody or antigen-binding fragment thereof of any of claims 1001 to 1011, comprising a binding domain comprising: [The present invention 1013] The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:25, (b) HC-CDR2 comprising SEQ ID NO:27, (c) HC-CDR3 comprising SEQ ID NO:29, (d) LC-CDR1 comprising SEQ ID NO:50, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1001 to 1012 of the present invention, comprising: [The present invention 1014] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:12 or SEQ ID NO:13; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:19; (c) a VH comprising SEQ ID NO: 12 and a VL comprising SEQ ID NO: 18; and / or (d) a VH domain comprising SEQ ID NO:13 and a VL domain comprising SEQ ID NO:19 1013. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [The present invention 1015] The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:33, (b) HC-CDR2 comprising SEQ ID NO:34, (c) HC-CDR3 comprising SEQ ID NO:36, (d) LC-CDR1 comprising SEQ ID NO:58, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1001 to 1012 of the present invention, comprising: [The present invention 1016] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:14; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:20; and / or (c) a VH domain comprising SEQ ID NO:14 and a VL domain comprising SEQ ID NO:20 1015. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [The present invention 1017] The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:38, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:41, (d) LC-CDR1 comprising SEQ ID NO:60, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. The isolated antibody or antigen-binding fragment thereof of any one of 1001 to 1012 of the present invention, comprising: [The present invention 1018] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:15; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:21; and / or (c) a VH domain comprising SEQ ID NO:15 and a VL domain comprising SEQ ID NO:21 1017. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [The present invention 1019] The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:43, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:41, (d) LC-CDR1 comprising SEQ ID NO:62, (e) LC-CDR22 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1001 to 1012 of the present invention, comprising: [The present invention 1020] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:16; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:22; and / or (c) a VH domain comprising SEQ ID NO: 16 and a VL domain comprising SEQ ID NO: 22 1019. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [The present invention 1021] The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:43, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:47, (d) LC-CDR1 comprising SEQ ID NO:63, (e) LC-CDR2 comprising SEQ ID NO:64, and (f) LC-CDR3 comprising SEQ ID NO:54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1001 to 1012 of the present invention, comprising: [The present invention 1022] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:17; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:23; and / or (c) a VH domain comprising SEQ ID NO:17 and a VL domain comprising SEQ ID NO:23 1021. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [The present invention 1023] A nucleic acid molecule encoding the CDR of the heavy chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of any one of the present inventions 1011 to 1022. [The present invention 1024] A nucleic acid molecule encoding the CDR of the light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of any one of the present inventions 1011 to 1022. [The present invention 1025] A cloning vector or expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of the present invention 1023 and / or the present invention 1024. [The present invention 1026] A cell comprising at least one of a nucleic acid molecule encoding an antibody or an antigen-binding fragment thereof that specifically binds to human maturation factor D of the present invention 1023 or the present invention 1024. [The present invention 1027] A method for producing an isolated antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, comprising culturing a cell of the present invention 1026 under conditions that allow expression of a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, and isolating the anti-maturation factor D-specific antibody or antigen-binding fragment thereof. [The present invention 1028] A composition comprising an antibody or an antigen-binding fragment thereof that specifically binds to human maturation factor D of any one of the present inventions 1001 to 1022. [The present invention 1029] A substrate for use in immunoassays, comprising at least one antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of any one of the present inventions 1001 to 1022. [The present invention 1030] A kit for detecting the presence or amount of maturation factor D in a test sample, comprising (a) at least one container and (b) at least one antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of any of the present inventions 1001 to 1022. [The present invention 1031] An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human factor D, wherein the epitope comprises or consists of "APPRGR" (SEQ ID NO:4). [The present invention 1032] 1031. An isolated antibody or antigen-binding fragment thereof of the present invention, which specifically binds to human pro-factor D (SEQ ID NO:2) and does not bind to mature factor D (SEQ ID NO:3). [The present invention 1033] The isolated antibody or antigen-binding fragment thereof of the present invention 1031 or 1032, wherein said antibody is a monoclonal antibody. [The present invention 1034] The isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1033, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. [This invention 1035] An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1034 of the present invention, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. [The present invention 1036] An isolated antibody or antigen-binding fragment thereof according to any one of 1031 to 1035 of the present invention, which is a single-chain molecule. [This invention 1037] 1037. The isolated antibody or antigen-binding fragment thereof of any of claims 1031 to 1036, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. [The present invention 1038] K<10 nM D An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1037 of the present invention, which binds to human profactor D at [This invention 1039] 1038. The isolated antibody or antigen-binding fragment thereof of any of claims 1031 to 1038, which is labeled with a detectable moiety. [The present invention 1040] An isolated antibody or antigen-binding fragment thereof according to any one of 1031 to 1039 of the present invention, which is immobilized on a substrate. [This invention 1041] An isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D of any of the present inventions 1031 to 1039, wherein the antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, and the CDRs are numbered according to the Kabat numbering system. [The present invention 1042] An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1041 of the present invention, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 139, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 145. [This invention 1043] The six CDRs are: (a) HC-CDR1, which comprises the amino acid sequence XYWMS (SEQ ID NO:201), where X at position 1 is N, S, or T; (b) the amino acid sequence (c) HC-CDR2 comprising the amino acid sequence AWFAX (SEQ ID NO: 203) (wherein X at position 5 is S, Y, or N); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence WASTRES (SEQ ID NO: 178); and (f) an LC-CDR3 comprising the amino acid sequence LQYYXYPYT (SEQ ID NO: 205) (wherein X at position 5 is T or S). 1043. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1042, comprising a binding domain comprising: [This invention 1044] The following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 149 or SEQ ID NO: 155; (b) HC-CDR2 comprising SEQ ID NO: 151 or SEQ ID NO: 156; (c) HC-CDR3 comprising SEQ ID NO: 153; (d) LC-CDR1 comprising SEQ ID NO: 176; (e) LC-CDR2 comprising SEQ ID NO: 178; and (f) LC-CDR3 comprising SEQ ID NO: 180. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1043, comprising a binding domain comprising: [This invention 1045] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:136; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:137; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:142; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:143; (e) a VH domain comprising SEQ ID NO: 136 and a VL domain comprising SEQ ID NO: 142; and / or (f) a VH domain comprising SEQ ID NO: 137 and a VL domain comprising SEQ ID NO: 143 1044. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [The present invention 1046] The six CDRs are: (a) HC-CDR1 comprising SEQ ID NO: 158; (b) HC-CDR2 comprising SEQ ID NO: 159 or SEQ ID NO: 163; (c) HC-CDR3 comprising SEQ ID NO: 161 or SEQ ID NO: 165; (d) LC-CDR1 comprising SEQ ID NO: 184 or SEQ ID NO: 189; (e) LC-CDR2 comprising SEQ ID NO: 178; and (f) LC-CDR3 comprising SEQ ID NO: 187. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1043, comprising a binding domain comprising: [This invention 1047] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:138; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:139; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:144; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:145; (d) a VH domain comprising SEQ ID NO: 138 and a VL domain comprising SEQ ID NO: 144; and / or (e) a VH domain comprising SEQ ID NO: 139 and a VL domain comprising SEQ ID NO: 145 1046. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [This invention 1048] An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1041 of the present invention, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NO: 140 and SEQ ID NO: 141, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO: 146 and SEQ ID NO: 147. [This invention 1049] The six CDRs are: (a) CDR-H1 comprising SEQ ID NO:167; (b) CDR-H2 comprising SEQ ID NO:169 or SEQ ID NO:173; (c) CDR-H3 comprising SEQ ID NO:171 or SEQ ID NO:174; (d) CDR-L1 comprising SEQ ID NO:194; (e) CDR-L2 comprising SEQ ID NO:196 or SEQ ID NO:199; and (f) CDR-L3 comprising SEQ ID NO:198 or SEQ ID NO:200. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1031 to 1041 of the present invention, comprising a binding domain comprising: [The present invention 1050] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:140; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:141; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:146; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:147; (e) a VH domain comprising SEQ ID NO:140 and a VL domain comprising SEQ ID NO:146; and / or (f) a VH domain comprising SEQ ID NO: 141 and a VL domain comprising SEQ ID NO: 147 1049. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [This invention 1051] A nucleic acid molecule encoding the CDR of the heavy chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human profactor D of any of 1031 to 1050 of the present invention. [This invention 1052] A nucleic acid molecule encoding the CDR of the light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human profactor D of any of 1031 to 1050 of the present invention. [This invention 1053] A cloning vector or expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human profactor D of the present invention 1051 and / or the present invention 1052. [This invention 1054] A cell comprising at least one of the nucleic acid molecules encoding the antibody or antigen-binding fragment thereof of the present invention 1051 and / or the present invention 1052 that specifically binds to human profactor D. [This invention 1055] A method for producing an isolated antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, comprising culturing a cell of the invention 1054 under conditions that allow expression of a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, and isolating the anti-pro-factor D-specific antibody or antigen-binding fragment thereof. [The present invention 1056] A composition comprising an antibody or an antigen-binding fragment thereof that specifically binds to human profactor D of any of the present inventions 1031 to 1050. [This invention 1057] A substrate for use in an immunoassay, comprising at least one antibody or antigen-binding fragment thereof that specifically binds to human profactor D of any of 1031 to 1050 of the present invention. [This invention 1058] A kit for detecting the presence or amount of pro-factor D in a test sample, comprising (a) at least one container and (b) at least one antibody or antigen-binding fragment thereof that specifically binds to human mature factor D of any of the present inventions 1031 to 1050. [This invention 1059] An isolated antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D, wherein the antibody comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85 to 88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89 to 93, and the CDRs are numbered according to the Kabat numbering system. [The present invention 1060] 1059. The isolated antibody or antigen-binding fragment thereof of the present invention, wherein said antibody is a monoclonal antibody. [The present invention 1061] The isolated antibody or antigen-binding fragment thereof of 1059 or 1060, wherein said antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. [The present invention 1062] An isolated antibody or antigen-binding fragment thereof according to any one of claims 1059 to 1061 of the present invention, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. [This invention 1063] 1063. The isolated antibody or antigen-binding fragment thereof of any of claims 1059 to 1062, wherein the antibody is a single-chain molecule. [This invention 1064] The isolated antibody or antigen-binding fragment thereof of any of claims 1059 to 1063, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. [This invention 1065] K<10 nM DAn isolated antibody or antigen-binding fragment thereof according to any one of claims 1059 to 1064 of the present invention, which binds to human factor D at [The present invention 1066] 1066. The isolated antibody or antigen-binding fragment thereof of any of claims 1059 to 1065, which is labeled with a detectable moiety. [This invention 1067] An isolated antibody or antigen-binding fragment thereof according to any one of 1059 to 1066 of the present invention, which is immobilized on a substrate. [The present invention 1068] An isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D of any of SEQ ID NOs. 1059 to 1067 of the present invention, wherein the antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, and the CDRs are numbered according to the Kabat numbering system. [This invention 1069] The following six CDRs: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO:95; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO:97; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO:99; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO:111; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO:113; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO:115. 1069. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1059 to 1068, comprising a binding domain comprising: [The present invention 1070] The six CDRs are: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO: 101; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO: 103 or 107; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO: 105 or 108; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO: 60 or 123; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO: 119, 124, or 126; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO: 121 or 125. 1069. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1059 to 1068, comprising a binding domain comprising: [This invention 1071] (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:85; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:86; (c) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:87; (d) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:88; (e) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:89; (f) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:90; (g) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:91; (h) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:92; (i) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:93; (j) a VH domain comprising SEQ ID NO:85 and a VL domain comprising SEQ ID NO:89 or SEQ ID NO:90; (k) a VH domain comprising SEQ ID NO:86 and a VL domain comprising SEQ ID NO:91; (l) a VH domain comprising SEQ ID NO:87 and a VL domain comprising SEQ ID NO:92; and / or (m) a VH domain comprising SEQ ID NO:88 and a VL domain comprising SEQ ID NO:93 1070. An isolated antibody or antigen-binding fragment thereof of the present invention, comprising at least one of: [This invention 1072] A nucleic acid molecule encoding the CDR of the heavy chain variable region of an antibody or an antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D of any one of 1059 to 1071 of the present invention. [This invention 1073] A nucleic acid molecule encoding the CDR of the light chain variable region of an antibody or an antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D of any one of 1059 to 1071 of the present invention. [This invention 1074] A cloning vector or expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that binds to an epitope common to human mature factor D and human pro-factor D of the present invention 1072 and / or 1073. [This invention 1075] A cell comprising at least one of the nucleic acid molecules encoding the antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human pro-factor D of the present invention 1072 and / or the present invention 1073. [This invention 1076] A method for producing an isolated antibody or antigen-binding fragment thereof that binds to an epitope common to human mature factor D and human pro-factor D, comprising culturing a cell of the present invention 1075 under conditions that allow expression of a nucleic acid molecule encoding the antibody or antigen-binding fragment thereof that binds to factor D, and isolating the anti-factor D-specific antibody or antigen-binding fragment thereof. [This invention 1077] A composition comprising an antibody or an antigen-binding fragment thereof that specifically binds to an epitope common to human maturation factor D and human profactor D according to any one of claims 1059 to 1071 of the present invention. [This invention 1078] A substrate for use in immunoassays, comprising at least one antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D of any one of 1059 to 1071 of the present invention. [This invention 1079] A kit for detecting the presence or amount of factor D in a test sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that binds to an epitope common to human mature factor D and human profactor D of any one of the present inventions 1059 to 1071. [The present invention 1080] 1. A kit comprising at least one monoclonal antibody that specifically detects or quantifies human maturation factor D (SEQ ID NO:3) and / or pro-factor D (SEQ ID NO:2) in an immunoassay, wherein the at least one monoclonal antibody is (i) a maturation factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope encompassing the amino terminus of human maturation factor D, wherein the epitope comprises or consists of the amino acid sequence ILGGREA (SEQ ID NO:5), and the antibody does not bind to human pro-factor D (SEQ ID NO:2); and / or (ii) A pro-Factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human Factor D, wherein the epitope comprises or consists of "APPRGR" (SEQ ID NO:4), and the antibody does not bind to mature Factor D (SEQ ID NO:3). Includes a kit. [This invention 1081] The kit of the present invention 1080, further comprising an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D (SEQ ID NO:3) and human pro-factor D (SEQ ID NO:2). [This invention 1082] The kit of invention 1080 or 1081, further comprising at least one container. [This invention 1083] Any of the kits of the present inventions 1080 to 1082, wherein the antibody or antigen-binding fragment thereof of subpart (i) that specifically binds to human maturation factor D comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, and the CDRs are numbered according to the Kabat numbering system. [This invention 1084] Any of the kits of the present inventions 1080 to 1083, wherein the antibody or antigen-binding fragment of subpart (ii) that specifically binds to an epitope in the activation ("pro") peptide of human factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, the CDRs being numbered according to the Kabat numbering system. [This invention 1085] The kit of any one of claims 1080 to 1084, wherein the immunoassay is an enzyme-linked immunosorbent assay (ELISA). [The present invention 1086] 1080. The kit of the present invention, wherein the mature factor D-specific antibody or antigen-binding fragment thereof of subpart (i) is a coating antibody. [This invention 1087] 1080. The kit of the present invention, wherein the mature factor D-specific antibody or antigen-binding fragment thereof of subpart (i) is a detection antibody. [This invention 1088] The kit of claim 1080, wherein the pro-factor D-specific antibody or antigen-binding fragment thereof of subpart (ii) is a coating antibody. [This invention 1089] 1080. The kit of claim 1080, wherein the pro-factor D-specific antibody or antigen-binding fragment thereof of subpart (ii) is a detection antibody. [The present invention 1090] A kit of the present invention 1081, wherein the anti-factor D antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85 to 88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89 to 93, and the CDRs are numbered according to the Kabat numbering system. [This invention 1091] 1. A method for determining the presence or amount of mature factor D in a test sample, comprising: (a) contacting a test sample with a mature factor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence, absence or amount of said antibody or antigen-binding fragment thereof bound to maturation factor D, wherein the presence of said binding indicates the presence or amount of maturation factor D in said sample. Including, The method, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof binds to an epitope in the N-terminal region of maturation factor D represented by amino acids ILGGREA (SEQ ID NO:5). [This invention 1092] 1092. The method of claim 1091, wherein the antibody or antigen-binding fragment thereof specifically binds to human mature factor D (SEQ ID NO:3) and does not bind to human pro-factor D (SEQ ID NO:2). [This invention 1093] 13. The method of claim 1091 or 1092, wherein said anti-human maturation factor D-specific antibody or antigen-binding fragment thereof is immobilized on a substrate. [This invention 1094] The method of any one of claims 1091 to 1093, wherein said immunoassay is an ELISA assay. [This invention 1095] Any of the methods of claims 1091 to 1094, wherein the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof is labeled with a detectable moiety, and step (b) comprises detecting the presence or amount of the detectable moiety. [This invention 1096] Any of the methods of claims 1091 to 1094, wherein the anti-human maturation factor D-specific antibody or its antigen-binding fragment is naked (i.e., unlabeled), and the presence or amount of the antibody or its fragment bound to maturation factor D is detected using a labeled antibody that binds to the anti-maturation factor D antibody. [This invention 1097] Any of the methods of claims 1091 to 1096, wherein the anti-human maturation factor D specific antibody or its antigen-binding fragment is immobilized (i.e., captured / coated) on a substrate, and the bound maturation factor D is detected using a second antibody that binds to a different epitope of factor D. [This invention 1098] 8. The method of any of claims 1091 to 1097, wherein the test sample is a biological sample obtained from a mammalian subject, for example, the biological sample is selected from the group consisting of blood, serum, plasma, urine, and cerebrospinal fluid. [This invention 1099] The method of claim 1098, wherein said mammalian subject is suffering from or at risk of developing an alternative pathway disease or disorder. [The present invention 1100] The method of claim 1098 or 1099, wherein the mammalian subject has been treated with a complement inhibitor, such as an alternative complement pathway inhibitor, such as a pro-factor D maturation inhibitor, such as a MASP-3 inhibitory antibody or an antigen-binding fragment thereof. [The present invention 1101] Any of the methods of the present inventions 1091 to 1100, wherein the anti-human maturation factor D-specific antibody or its antigen-binding fragment comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23. [The present invention 1102] The anti-human maturation factor D-specific antibody or antigen-binding fragment thereof has the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO: 65), where X at position 1 is T, I, or S, and X at position 3 is G or I; (b) amino acid sequence HC-CDR2 containing TIFF2025076475000009.tif4128 (wherein X at position 11 is H or N, and X at position 16 is S or R); (c) amino acid sequence HC-CDR3 containing TIFF2025076475000010.tif4128 (wherein X at position 6 is R, G, or N, X at position 7 is S or Y, X at position 8 is F, I, or V, and X at position 10 is D or H); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69) (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54). 1102. The method of any of claims 1091 to 1101, comprising a binding domain comprising: [The present invention 1103] The anti-human maturation factor D specific antibody or antigen-binding fragment thereof has the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:25, (b) HC-CDR2 comprising SEQ ID NO:27, (c) HC-CDR3 comprising SEQ ID NO:29, (d) LC-CDR1 comprising SEQ ID NO:50, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. The method of claim 1102, comprising a binding domain comprising: [The present invention 1104] 1. A method for determining the presence or amount of pro-factor D in a test sample, comprising: (a) contacting a test sample with an anti-human profactor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or amount of said antibody or antigen-binding fragment thereof bound to pro-factor D, wherein the presence of said binding indicates the presence or amount of pro-factor D in said sample. Including, A method wherein the anti-human mature pro-factor D specific antibody or antigen-binding fragment thereof specifically binds to an epitope in the activation ("pro") peptide of human factor D designated as "APPRGR" (SEQ ID NO:4). [This invention 1105] 1105. The method of claim 1104, wherein said antibody or antigen-binding fragment thereof specifically binds to human pro-factor D (SEQ ID NO:2) and does not bind to human mature factor D (SEQ ID NO:3). [The present invention 1106] 1106. The method of claim 1104 or 1105, wherein said anti-human profactor D-specific antibody or antigen-binding fragment thereof is immobilized on a substrate. [This invention 1107] The method of any one of claims 1104 to 1106, wherein said immunoassay is an ELISA assay. [This invention 1108] Any of the methods of claims 1104 to 1107, wherein the anti-human profactor D-specific antibody or antigen-binding fragment thereof is labeled with a detectable moiety, and step (b) comprises detecting the presence or amount of the detectable moiety. [This invention 1109] Any of the methods of claims 1104 to 1107, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof is naked (i.e., unlabeled), and the presence or amount of the antibody or fragment thereof bound to mature factor D is detected using a labeled antibody that binds to the anti-pro-factor D antibody. [The present invention 1110] Any of the methods of claims 1104 to 1109, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof is immobilized (i.e., captured / coated) on a substrate, and the bound pro-factor D is detected using a second antibody that binds to a different epitope of factor D. [The present invention 1111] The method of any of claims 1104 to 1110, wherein the test sample is a biological sample obtained from a mammalian subject, for example, the biological sample is selected from the group consisting of blood, serum, plasma, urine, and cerebrospinal fluid. [The present invention 1112] The method of claim 1111, wherein said mammalian subject is suffering from or at risk of developing an alternative pathway disease or disorder. [The present invention 1113] The method of invention 1111 or 1112, wherein the mammalian subject has been treated with a complement inhibitor, such as an alternative complement pathway inhibitor, such as a pro-factor D maturation inhibitor, such as a MASP-3 inhibitory antibody or its antigen-binding fragment. [This invention 1114] Any of the methods of claims 1104 to 1113, wherein the anti-human profactor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR-1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, and the CDRs are numbered according to the Kabat numbering system. [This invention 1115] Any of the methods of claims 1104 to 1113, wherein the anti-human profactor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 139, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 145, and the CDRs are numbered according to the Kabat numbering system. [The present invention 1116] Any of the methods of claims 1104 to 1113, wherein the anti-human profactor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NO: 140 and SEQ ID NO: 141, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO: 146 and SEQ ID NO: 147, and the CDRs are numbered according to the Kabat numbering system. [This invention 1117] The anti-human pro-factor D specific antibody or antigen-binding fragment thereof comprises the following six CDRs: (a) CDR-H1 comprising SEQ ID NO: 167, (b) CDR-H2 comprising SEQ ID NO: 169 or SEQ ID NO: 173, (c) CDR-H3 comprising SEQ ID NO: 171 or SEQ ID NO: 174, (d) CDR-L1 comprising SEQ ID NO: 194, (e) CDR-L2 comprising SEQ ID NO: 196 or SEQ ID NO: 199, and (f) CDR-L3 comprising SEQ ID NO: 198 or SEQ ID NO: 200. The method of any one of claims 1104 to 1113, comprising a binding domain comprising: [This invention 1118] 1. A method for assessing the degree of alternative pathway complement (APC) activation in a test sample, comprising: (a) providing a test sample; (b) performing an immunoassay comprising at least one of the following: (i) capturing and detecting mature factor D in the test sample, wherein the mature factor D is captured or detected with a mature factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but not pro-factor D; and / or (ii) capturing and detecting pro-factor D in the test sample, wherein the pro-factor D is captured or detected with a pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but does not bind to mature factor D; and (c) comparing the level of mature factor D detected according to (b)(i) with a predetermined level or a control sample and / or comparing the level of pro-factor D detected according to (b)(ii) with a predetermined level or a control sample, wherein the detected level of mature factor D and / or pro-factor D in the test sample indicates the degree of alternative pathway complement activation. A method comprising: [This invention 1119] The method of the present invention 1118, wherein step (b)(i) comprises capturing mature factor D with a mature factor D-specific monoclonal antibody or its antigen-binding fragment that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but not to pro-factor D, and detecting with an antibody or its antigen-binding fragment that binds to an epitope common to both human mature factor D and human pro-factor D. [The present invention 1120] The method of the present invention 1118, wherein step (b)(i) comprises capturing mature factor D with an anti-factor D antibody or its antigen-binding fragment that binds to an epitope common to both human mature factor D and human pro-factor D, and detecting with a mature factor D-specific monoclonal antibody or its antigen-binding fragment that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but not pro-factor D. [This invention 1121] Any of the methods of claims 1118 to 1120, wherein step (b)(ii) comprises capturing pro-factor D with a pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but not mature factor D, and detecting with an antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D. [This invention 1122] Any of the methods of claims 1118 to 1120, wherein step (b)(ii) comprises capturing pro-factor D with an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, and detecting with a monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but does not bind to mature factor D. [This invention 1123] Any of the methods of the present inventions 1118 to 1122, wherein the mature factor D-specific monoclonal antibody or its antigen-binding fragment that specifically binds to an epitope in "ILGGREA" (SEQ ID NO: 5) present in mature factor D but does not bind to pro-factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, and the CDRs are numbered according to the Kabat numbering system. [This invention 1124] Any of the methods of inventions 1118 to 1123, wherein the pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO: 4) present in pro-factor D but does not bind to mature factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141 and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, the CDRs being numbered according to the Kabat numbering system. [This invention 1125] 1125. The method of any one of claims 1118 to 1124, wherein said test sample is a biological sample obtained from a mammalian subject. [Invention 1126] 1125. The method of claim 1125, wherein said biological sample comprises whole blood, serum, plasma, urine, or cerebrospinal fluid. [This invention 1127] 1127. The method of any of claims 1118 to 1126, wherein said test sample comprises a complement inhibitor, eg, an alternative complement pathway inhibitor, eg, a pro-factor D maturation inhibitor, eg, a MASP-3 inhibitor. [This invention 1128] The method of the present invention 1125, wherein the mammalian subject has been treated with a complement inhibitor, e.g., an alternative complement pathway inhibitor, e.g., a pro-factor D maturation inhibitor, e.g., a MASP-3 inhibitor, and the degree of inhibition of the alternative pathway is measured using the assay. [This invention 1129] 1129. The method of any one of claims 1125 to 1128, wherein said mammalian subject is a human subject. [The present invention 1130] The method of claim 1129, wherein said human subject is suffering from, at risk of developing, or suspected of having an alternative pathway disease or disorder. [This invention 1131] The alternative pathway disease or disorder is selected from the group consisting of paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD, and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, and polyposis. 1130. The method of the present invention, wherein the patient is selected from the group consisting of: idiopathic sclerosis, Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft-versus-host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis. [This invention 1132] The method of claim 1128, wherein the control sample is a sample taken from the subject before treatment with a MASP-3 inhibitor or a sample taken at an earlier time point during the course of treatment with a MASP-3 inhibitor. [This invention 1133] The method of the present invention 1128 or 1132, wherein the MASP-3 inhibitor is a MASP-3 inhibitory antibody or its antigen-binding fragment. [This invention 1134] The MASP-3 inhibitory antibody is a monoclonal antibody or an antigen-binding fragment thereof that binds to MASP-3; and (i) a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 220, 222, 223, 225, 226, and 228, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 221, 224, and 227, wherein the CDRs are numbered according to the Kabat numbering system; (ii) HC-CDR1 comprising SEQ ID NO:229 (TDDIN); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 236 (LEDTY), an HC-CDR3 comprising SEQ ID NO: 236 (LEDTY), and a light chain variable region comprising LC-CDR1 comprising TIFF2025076475000013.tif4128, LC-CDR2 comprising SEQ ID NO:178 (WASTRES), and LC-CDR3 comprising SEQ ID NO:242 (KQSYNLYT); (iii) HC-CDR1 comprising SEQ ID NO: 230 (SYGMS); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 237 (GGEAMDY); and a HC-CDR3 comprising SEQ ID NO: 237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising TIFF2025076475000015.tif4128, an LC-CDR2 comprising SEQ ID NO:241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO:243 (WQGTHFPWT); or (iv) HC-CDR1 comprising SEQ ID NO: 231 (GKWIE); TIFF2025076475000016.tif4128 or a heavy chain variable region comprising HC-CDR2 comprising SEQ ID NO: 238 (SEDV), and HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising LC-CDR1 comprising SEQ ID NO: 239, LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT). The method of the present invention 1133, comprising at least one of the following: [This invention 1135] 1. A method for monitoring the effectiveness of treatment with a MASP-3 inhibitory antibody or antigen-binding fragment thereof in a mammalian subject, comprising: (a) administering a dose of a MASP-3 inhibitory antibody or antigen-binding fragment thereof to a mammalian subject at a first time point; (b) assessing a first concentration of mature factor D and / or pro-factor D in a biological sample obtained from the subject after step (a); (c) treating the subject with the MASP-3 inhibitory antibody or antigen-binding fragment thereof at a second time point; (d) assessing a second concentration of mature factor D and / or pro-factor D in a biological sample obtained from the subject after step (c); and (e) comparing the level of mature factor D and / or pro-factor D assessed in step (b) with the level of mature factor D and / or pro-factor D assessed in step (d) to determine the efficacy of the MASP-3 inhibitory antibody or antigen-binding fragment thereof in the mammalian subject. A method comprising: [This invention 1136] The method of the present invention 1135 further comprises a step of adjusting the dose of the MASP-3 inhibitory antibody or its antigen-binding fragment. [This invention 1137] The method of the present invention 1136, wherein the dose of the MASP-3 inhibitory antibody or its antigen-binding fragment administered to the subject is increased if the level of the maturation factor D is higher than the control or reference standard. [This invention 1138] The method of the present invention 1136, wherein the dose of the MASP-3 inhibitory antibody or its antigen-binding fragment administered to the subject is increased if the level of pro-factor D is lower than the control or reference standard. [This invention 1139] The method of the present invention 1137 or 1138, wherein when an increased dose of the MASP-3 inhibitory antibody or its antigen-binding fragment is administered to the subject, steps (b) to (e) are repeated to determine whether the increased dose is sufficient to adjust the levels of mature factor D and / or pro-factor D to the desired level compared to the respective control or reference standard. [This invention 1140] 1135. The method of claim 1135, wherein steps (b) and (d) comprise assessing the concentration of mature factor D in said biological sample by immunoassay. [This invention 1141] The method of claim 1140, wherein the immunoassay comprises (i) a first monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope comprising or consisting of amino acids ILGGREA (SEQ ID NO:5) in the N-terminal region of human maturation factor D, but does not bind to human pro-factor D, and (ii) a second antibody or antigen-binding fragment thereof that binds to an epitope common to both human maturation factor D and human pro-factor D, wherein the first and second antibodies or antigen-binding fragments function together in the immunoassay to specifically detect or quantify the amount of mature factor D protein (SEQ ID NO:3) but not pro-factor D protein (SEQ ID NO:2) that may be present in the biological sample. [This invention 1142] 1135. The method of claim 1135, wherein steps (b) and (d) comprise assessing the concentration of pro-factor D in said biological sample by immunoassay. [This invention 1143] The method of claim 1142, wherein the immunoassay comprises (i) a first monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope comprising or consisting of amino acids APPRGR (SEQ ID NO:4) in the propeptide of human factor D, but does not bind to human mature factor D, and (ii) a second antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the first and second antibodies or antigen-binding fragments function together in the immunoassay to specifically detect or quantify the amount of pro-factor D protein (SEQ ID NO:2) but not mature factor D protein (SEQ ID NO:3) that may be present in the biological sample. [This invention 1144] 1144. The method of any one of claims 1135 to 1143, wherein said mammalian subject is a human subject. [Invention 1145] The method of claim 1144, wherein said human subject is suffering from or at risk of developing an alternative pathway disease or disorder. [Invention 1146] The alternative pathway disease or disorder is selected from the group consisting of paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD, and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, and polyposis. 1145. The method of the present invention, wherein the patient is selected from the group consisting of: idiopathic sclerosis, Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft-versus-host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis. [This invention 1147] The method of the present invention 1135, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding fragment. [Invention 1148] The MASP-3 inhibitory antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof that binds to MASP-3; and (i) a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 220, 222, 223, 225, 226, and 228, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 221, 224, and 227, wherein the CDRs are numbered according to the Kabat numbering system; (ii) HC-CDR1 comprising SEQ ID NO:229 (TDDIN); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 236 (LEDTY); and a HC-CDR3 comprising SEQ ID NO: 236 (LEDTY). a light chain variable region comprising LC-CDR1 comprising TIFF2025076475000019.tif5128, LC-CDR2 comprising SEQ ID NO:178 (WASTRES), and LC-CDR3 comprising SEQ ID NO:242 (KQSYNLYT); (iii) HC-CDR1 comprising SEQ ID NO: 230 (SYGMS); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 237 (GGEAMDY); and a HC-CDR3 comprising SEQ ID NO: 237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising TIFF2025076475000021.tif4128, an LC-CDR2 comprising SEQ ID NO:241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO:243 (WQGTHFPWT); or (iv) HC-CDR1 comprising SEQ ID NO: 231 (GKWIE); TIFF2025076475000022.tif4128 or a heavy chain variable region comprising HC-CDR2 comprising SEQ ID NO: 238 (SEDV), and HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising LC-CDR1 comprising SEQ ID NO: 239, LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT). The method of the present invention 1147, comprising at least one of the following: [This invention 1149] 1. A method of treating a mammalian subject suffering from or at risk of developing an alternative pathway disease or disorder, comprising administering to the subject: (i) the level of pro-factor D in one or more samples taken from the subject is low or decreased compared to a predetermined pro-factor D level or compared to the level of pro-factor D in one or more control samples; and / or (ii) the level of maturation factor D in one or more samples taken from the subject is elevated or increased compared to the predetermined level of maturation factor D or compared to the level of maturation factor D in one or more control samples. A method comprising the step of administering an MASP-3 inhibitory antibody or its antigen-binding fragment to the subject when the subject is determined to have MASP-3. [This invention 1150] 1149. The method of claim 1149, wherein the level of pro-factor D in one or more samples taken from said subject is determined by performing an immunoassay comprising the use of a pro-factor D-specific monoclonal antibody. [This invention 1151] The method of claim 1150, wherein the immunoassay comprises (i) a first monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope comprising or consisting of amino acids APPRGR (SEQ ID NO:4) in the propeptide of human factor D, but does not bind to human mature factor D, and (ii) a second antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the first and second antibodies or antigen-binding fragments function together in the immunoassay to specifically detect or quantify the amount of pro-factor D protein (SEQ ID NO:2) but not mature factor D protein (SEQ ID NO:3) that may be present in the sample. [This invention 1152] 1149. The method of claim 1149, wherein the level of mature factor D in one or more samples taken from the subject is determined by performing an immunoassay comprising the use of a mature factor D-specific monoclonal antibody or an antigen-binding fragment thereof. [This invention 1153] 1152. The method of claim 1152, wherein the immunoassay comprises: (i) a first monoclonal antibody, or antigen-binding fragment thereof, that specifically binds to an epitope comprising or consisting of amino acids ILGGREA (SEQ ID NO:5) in the N-terminal region of human maturation factor D, but not human pro-factor D; and (ii) a second antibody, or antigen-binding fragment thereof, that binds to an epitope common to both human maturation factor D and human pro-factor D, wherein the first and second antibodies, or antigen-binding fragments thereof, function together in the immunoassay to specifically detect or quantify the amount of mature factor D protein (SEQ ID NO:3) but not pro-factor D protein (SEQ ID NO:2) that may be present in the sample. [This invention 1154] 1154. The method of any of claims 1149 to 1153, wherein said mammalian subject is a human subject. [Invention 1155] The human subject is a patient suffering from paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (AMD, including wet AMD and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple sclerosis, Guillain-Barré syndrome, Alzheimer's disease, or Alzheimer's disease. 1154. The method of claim 1154, wherein the patient suffers from or is at risk of developing an alternative pathway disease or disorder selected from the group consisting of: myeloma, myeloma syndrome, myeloma stenosis, myeloma fibroelastosis, myeloma fibroelastosis (MFS), myeloma fibroelastosis (MFD ... [Invention 1156] The method of the present invention 1149, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding fragment. [This invention 1157] The MASP-3 inhibitory antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof that binds to MASP-3; and (i) a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 220, 222, 223, 225, 226, and 228, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 221, 224, and 227, wherein the CDRs are numbered according to the Kabat numbering system; (ii) HC-CDR1 comprising SEQ ID NO:229 (TDDIN); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 236 (LEDTY); and a HC-CDR3 comprising SEQ ID NO: 236 (LEDTY). a light chain variable region comprising LC-CDR1 comprising TIFF2025076475000025.tif5128, LC-CDR2 comprising SEQ ID NO:178 (WASTRES), and LC-CDR3 comprising SEQ ID NO:242 (KQSYNLYT); (iii) HC-CDR1 comprising SEQ ID NO: 230 (SYGMS); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 237 (GGEAMDY); and a HC-CDR3 comprising SEQ ID NO: 237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising TIFF2025076475000027.tif4128, an LC-CDR2 comprising SEQ ID NO:241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO:243 (WQGTHFPWT); or (iv) HC-CDR1 comprising SEQ ID NO: 231 (GKWIE); TIFF2025076475000028.tif4128 or a heavy chain variable region comprising HC-CDR2 comprising SEQ ID NO: 238 (SEDV), and HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising LC-CDR1 comprising SEQ ID NO: 239, LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT). The method of the present invention 1156, comprising at least one of the following: [This invention 1158] A pharmaceutical composition comprising a MASP-3 inhibitory antibody or its antigen-binding fragment in an aqueous solution comprising a buffer system having a pH of 6.0±5%, 20±5% mM histidine, 100±5% mg / mL sucrose, and 0.035%±5% polysorbate 80, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is contained at a concentration of 110 mg / mL±5%, and the MASP-3 inhibitory antibody or its antigen-binding fragment comprises an HC-CDR1 comprising SEQ ID NO:231 (GKWIE), TIFF2025076475000030.tif4128 or A pharmaceutical composition comprising a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 238 (SEDV), and an HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT). [This invention 1159] The pharmaceutical composition of the present invention 1158, which is sterile. [The present invention 1160] A pharmaceutical composition of the present invention 1158, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a heavy chain variable region that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to SEQ ID NO:226 or SEQ ID NO:227, and a light chain variable region that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to SEQ ID NO:227. [This invention 1161] A pharmaceutical composition of any of claims 1158 to 1160, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is selected from the group consisting of a human antibody, a humanized antibody, a chimeric antibody, a murine antibody, and an antigen-binding fragment of any of them. [This invention 1162] A pharmaceutical composition of any of claims 1158 to 1161, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is selected from the group consisting of a single-chain antibody, an ScFv, a Fab fragment, a Fab' fragment, an F(ab')2 fragment, a monovalent antibody lacking a hinge region, and a full-length antibody. [This invention 1163] A pharmaceutical composition of any of claims 1158 to 1162, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment further comprises an immunoglobulin constant region. [Invention 1164] The pharmaceutical composition of any of claims 1158 to 1163, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a human IgG4 constant region. [This invention 1165] A pharmaceutical composition of the present invention 1164, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a human IgG4 constant region having an S228P mutation. [Invention 1166] A pharmaceutical composition of the present invention 1164 or 1165, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a mutation that promotes FcRn interaction at low pH. [This invention 1167] A pharmaceutical composition of the present invention 1166, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a human IgG4 constant region shown as SEQ ID NO:245. [Invention 1168] An article of manufacture containing any one of the pharmaceutical compositions of the present inventions 1158 to 1167. [This invention 1169] The article of manufacture of the present invention 1168, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is in a unit dosage form of 10 mg to 1000 mg suitable for therapeutic administration to a human subject. [This invention 1170] The article of manufacture of the present invention 1169, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is in a unit dosage form of 100 mg to 200 mg suitable for therapeutic administration to a human subject. [This invention 1171] An article of manufacture according to any one of claims 1168 to 1170, comprising a container and a label or package insert on or associated with the container. [This invention 1172] 1171. The article of manufacture of claim 1171, wherein the container is selected from the group consisting of a bottle, an ampoule, a pouch (e.g., an intravenous infusion bag), a vial, a syringe, and a cartridge. [This invention 1173] A pharmaceutical composition of any of claims 1158-1167 or an article of manufacture of any of claims 1168-1172 for use in treating a subject suffering from or at risk of developing an alternative pathway disease or disorder. [This invention 1174] The alternative pathway disease or disorder is selected from the group consisting of paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD, and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple sclerosis, The pharmaceutical composition or article of manufacture of claim 1173, wherein the disease is selected from the group consisting of Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft-versus-host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis. [Brief explanation of the drawings]

[0022] The foregoing aspects and many of the attendant advantages of this invention will become more readily appreciated as the same become better understood by reference to the following detailed description, when taken in conjunction with the accompanying drawings, in which:

[0023] [Figure 1] 1 is a schematic diagram showing the classical, lectin, and alternative complement pathways. [Figure 2] The amino acid sequences of (i) human full-length Factor D (SEQ ID NO:1), including the signal sequence aa 1-19 (shown in italics) and the underlined activation (pro)peptide, are provided; (ii) human pro-Factor D (SEQ ID NO:2) with the underlined pro-peptide; and (iii) human mature Factor D (SEQ ID NO:3). [Figure 3] 1 provides an alignment of the amino acid sequences of profactor D from various species. [Figure 4]1 shows a graph of titration of antisera from representative mouse #2 after immunization with a peptide corresponding to the N-terminus of mature human Factor D, as described in Example 1. [Figure 5] 1 graphically depicts the results of a capture ELISA assay in which hybridoma supernatants were screened for binding to human mature factor D or human pro-factor D when captured by polyclonal anti-factor D antibody AF1824 (R&D Systems), as described in Example 1. [Figure 6A] The graph shows the results of an ELISA assay using coating polyclonal goat anti-human CFD 1824 to detect hybridoma supernatant 14A11 present in each condition described in Example 1. As shown in Figure 6A, hybridoma supernatant 14A11 can selectively detect recombinant mature complement factor D, but not recombinant pro-factor D, as described in Example 1. [Figure 6B] The graph shows the results of an ELISA assay using coating polyclonal goat anti-human CFD 1824 to detect hybridoma supernatant 6G6 present in each condition described in Example 1. As shown in Figure 6B, hybridoma supernatant 6G6 can selectively detect recombinant mature complement factor D, but not recombinant pro-factor D, as described in Example 1. [Figure 6C] The graph shows the results of an ELISA assay using coating polyclonal goat anti-human CFD 1824 to detect the presence of monoclonal antibody mAb1824 (R&D Systems) in each condition described in Example 1. As shown in Figure 6C, monoclonal antibody 1824 (R&D Systems) detects both recombinant mature CFD and recombinant active CFD, and therefore is unable to selectively detect mature CFD compared to pro-CFD, as described in Example 1. [Figure 7A]As described in Example 2, an amino acid alignment of heavy chain variable region (VH) sequences for anti-human maturation factor D specific clones is shown: 6G6_VH (SEQ ID NO:12), 14A11_VH (SEQ ID NO:13), 27B3_VH (SEQ ID NO:14), 58F5_VH (SEQ ID NO:15), 49G3_VH (SEQ ID NO:16), and 10G1_VH (SEQ ID NO:17). [Figure 7B] As described in Example 2, an amino acid alignment of the light chain variable region (VL) sequences for anti-human maturation factor D specific clones is shown: 6G6_VK (SEQ ID NO: 18), 14A11_VK: (SEQ ID NO: 19), 27B3_VK: (SEQ ID NO: 20), 58F5_VK: (SEQ ID NO: 21), 49G3_VK: (SEQ ID NO: 22), and 10G1_VK: (SEQ ID NO: 23). [Figure 8] 8 graphically depicts the detection (or lack thereof) of recombinant human pro-factor D or mature factor D using a number of candidate anti-human mature factor D-specific antibodies. As shown in FIG. 8, all of the purified antibodies tested, namely, 6G6, 14A11, 10G1, 49G3, 27B3, and 58F5, were found to be specific for the mature form of factor D, as described in Example 3. [Figure 9] 9 graphically depicts titration of serum from representative mouse #1189 after immunization with human mature factor D in the presence of recombinant mature factor D or recombinant pro-factor D. As shown in FIG. 9, serum from representative mouse #1189 contains antibodies capable of binding to both mature factor D and pro-factor D, as described in Example 4. [Figure 10A] As described in Example 5, an amino acid alignment of heavy chain variable region (VH) sequences for anti-human factor D clones is shown: 3C5_VH (SEQ ID NO:85), 30H2_VH (SEQ ID NO:85), 11H1_VH (SEQ ID NO:86), 12H10_VH (SEQ ID NO:87), and 7H2_VH (SEQ ID NO:88). [Figure 10B]As described in Example 5, an amino acid alignment of the light chain variable region (VL) sequences for anti-human factor D clones is shown: 3C5_VL (SEQ ID NO:89), 30H2_VL (SEQ ID NO:90), 11H1_VL (SEQ ID NO:91), 12H10_VL (SEQ ID NO:92) and 7H2_VL (SEQ ID NO:93). [Figure 11] Figure 11A graphically depicts the binding of recombinant human pro- or mature factor D to candidate anti-human factor D antibody 3C5, demonstrating that antibody 3C5 binds to both human pro- and mature factor D, as described in Example 5. Figure 11B graphically depicts the binding of recombinant human pro- or mature factor D to candidate anti-human factor D antibody 12H10, demonstrating that antibody 12H10 binds to both human pro- and mature factor D, as described in Example 5. [Figure 12] Figure 12A graphically shows the results of an ELISA assay in which recombinant anti-factor D antibody 3C5 was coated onto an ELISA plate to capture recombinant human and cynomolgus monkey mature and pro-factor D (huMat CFD, cyMat CFD, huPro CFD, and cyPro CFD). Samples of factor D-depleted human serum (CFD Dpl serum) and pooled normal cynomolgus monkey plasma (NCP) were also captured. Captured factor D was detected using the mouse IgG2a Fc version of the anti-human mature factor D-specific mAb 14A11, as described in Example 6. Figure 12B graphically shows the results of an ELISA assay in which recombinant anti-factor D antibody 12H10 was coated onto an ELISA plate to capture recombinant human and cynomolgus monkey mature and pro-factor D (huMat CFD, cyMat CFD, huPro CFD, and cyPro CFD). Samples of factor D-depleted human serum (CFD Dpl serum) and pooled normal cynomolgus monkey plasma (NCP) were also captured. Detection of captured factor D was performed using the mouse IgG2a Fc version of the anti-human mature factor D specific mAb 14A11, as described in Example 6. [Figure 13]1 is a graph showing the detection of human and cynomolgus mature factor D and pro-factor D using a combination of capture antibody 3C5 (anti-human / cynomolgus factor D) and detection antibody 14A11 (anti-human / cynomolgus mature factor D specific) in an ELISA assay, as described in Example 6. [Figure 14] 14 graphically depicts titration of serum from representative mouse #2 after immunization with a synthetic peptide corresponding to amino acid residues 26-32 of human complement factor D "ILGGREA" (SEQ ID NO:5) in the presence of recombinant mature factor D or recombinant pro-factor D. As shown in FIG. 14, serum from representative mouse #2 contains antibodies capable of selectively binding to mature factor D compared to pro-factor D, as described in Example 7. [Figure 15] Graph showing the results of a capture ELISA assay in which hybridoma supernatants were screened for binding to human pro-factor D or human mature factor D when captured by polyclonal anti-factor D antibody AF1824 (R&D Systems), as described in Example 7. [Figure 16A] As described in Example 8, an amino acid alignment of heavy chain variable region (VH) sequences for anti-human pro-factor D specific clones is shown: 18F5_VH (SEQ ID NO: 136), 1F9_VH (SEQ ID NO: 137), 2A4_VH (SEQ ID NO: 138), 20A1_VH (SEQ ID NO: 139), 13A10_VH (SEQ ID NO: 140) and 21H1_VH (SEQ ID NO: 141). [Figure 16B] As described in Example 8, an amino acid alignment of light chain variable region (VL) sequences for anti-human pro-factor D specific clones is shown: 18F5_VK (SEQ ID NO: 142), 1F9_VK (SEQ ID NO: 143), 2A4_VK (SEQ ID NO: 144), 20A1_VK (SEQ ID NO: 145), 13A10_VK (SEQ ID NO: 146), and 21H1_VK (SEQ ID NO: 147). [Figure 17]Figure 17A graphically depicts the detection of recombinant human pro-factor D using a number of candidate anti-human pro-factor D-specific antibodies. As shown in Figure 17A, all of the purified antibodies tested, i.e., 18F5, 1F9, 2A4, 20A1, 13A10, and 21H1, were able to detect the pro-form of factor D, as described in Example 9. Figure 17B graphically depicts the detection of recombinant human mature factor D using a number of candidate anti-human pro-factor D-specific antibodies. As shown in Figure 17B, none of the purified antibodies tested, i.e., 18F5, 1F9, 2A4, 20A1, 13A10, and 21H1, were able to detect the mature form of factor D, as described in Example 9. [Figure 18] 1 is a graph showing the detection of pro-factor D and mature factor D in an ELISA assay using anti-pro-factor D antibody 21H1 as the coating antibody and goat polyclonal anti-factor D antibody AF1824 (R&D Systems) as the detecting antibody, as described in Example 9. [Figure 19] Graph showing the detection of pro-factor D and mature factor D in normal human plasma (NHP), normal human serum (NHS), or factor D-depleted serum (Df-Dpl serum) in an ELISA assay using anti-pro-factor D antibody 21H1 as the coating antibody and goat polyclonal anti-factor D antibody AF1824 (R&D Systems) as the detection antibody, as described in Example 9. [Figure 20] Graph showing the amount of pro-factor D present in normal human serum (NHS), C1q-depleted serum (C1q-Dpl), factor D-depleted serum (Df-Dpl), and 3MC syndrome patient serum as determined by ELISA assay using anti-pro-factor D antibody 21H1 as the coating antibody and goat polyclonal anti-factor D antibody AF1824 (R&D Systems) as the detection antibody, as described in Example 9. [Figure 21]Figure 21A graphically depicts the amount of maturation factor D in cynomolgus monkeys over a 912 hour period following treatment with a representative anti-MASP-3 mAb 13B1, as described in Example 11. Figure 21B graphically depicts the standard curve as determined from a four-parameter logistic curve of cynomolgus monkey recombinant maturation factor D dilutions, as described in Example 11. [Figure 22] Figure 22A graphically depicts the concentration of mature factor D in monkeys over a 1344 hour period following subcutaneous (SC) or intravenous (IV) administration of anti-MASP-3 mAb 13B1, as measured by an ELISA assay using mature factor D-specific antibody 14A11, as described in Example 12. Figure 22B graphically depicts ex vivo alternative pathway activity (% of baseline) over a 1344 hour period following administration of anti-MASP-3 mAb 13B1, as determined by a factor Ba assay, as described in Example 12. [Figure 23] 10 graphically depicts the PD-PD relationship between the effect of anti-MASP-3 mAb 13B1 on ex vivo alternative pathway activity and mature factor D concentrations after a single intravenous bolus or subcutaneous administration in monkeys, as described in Example 12. [Figure 24] As described in Example 12, the PK-PD relationship between the dose of anti-MASP-3 mAb 13B1 and the effect on maturation factor D (% of baseline) is shown graphically. [Figure 25A] 1 graphically depicts subject serum concentrations of mAb13B1 over a period of up to 84 days following intravenous (IV) administration of mAb13B1, as determined by ELISA, as described in Example 13. [Figure 25B] The graph shows the levels of maturation factor D in subjects over a period of 84 days after intravenous (IV) administration of anti-MASP-3 mAb 13B1, as determined by an ELISA assay using the maturation factor D-specific antibody 14A11 as the detection antibody, as described in Example 13. DETAILED DESCRIPTION OF THE INVENTION

[0024] Sequence Listing Description SEQ ID NO: 1: Human full-length factor D amino acid sequence (including signal sequence) SEQ ID NO:2: Human profactor D amino acid sequence (without signal sequence) SEQ ID NO: 3: Human maturation factor D amino acid sequence SEQ ID NO: 4: Human propeptide "APPRGR" corresponding to residues 20-25 of human full-length factor D SEQ ID NO: 5: N-terminal peptide of human maturation factor D corresponding to residues 1-7 of human maturation factor D ("ILGGREA") SEQ ID NO: 6: Synthetic ILGGREA peptide-KLH conjugate TIFF2025076475000032.tif3128SEQ ID NO:7: Human MASP-3 protein SEQ ID NO: 8: Macaque full-length factor D SEQ ID NO: 9: Canis full-length factor D SEQ ID NO: 10: Rattus full-length factor D SEQ ID NO:11: Mus musculus full-length factor D Anti-human maturation factor D specific mAb: VH chain SEQ ID NO:12: mAb clone 6G6 VH amino acid sequence SEQ ID NO:13: mAb clone 14A11 VH amino acid sequence SEQ ID NO:14: mAb clone 27B3 VH amino acid sequence SEQ ID NO:15: mAb clone 58F5 VH amino acid sequence SEQ ID NO:16: mAb clone 49G3 VH amino acid sequence SEQ ID NO:17: mAb clone 10G1 VH amino acid sequence Anti-human maturation factor D specific mAb: VL chain SEQ ID NO:18: mAb clone 6G6 VL amino acid sequence SEQ ID NO:19: mAb clone 14A11 VL amino acid sequence SEQ ID NO:20: mAb clone 27B3 VL amino acid sequence SEQ ID NO:21: mAb clone 58F5 VL amino acid sequence SEQ ID NO:22: mAb clone 49G3 VL amino acid sequence SEQ ID NO:23: mAb clone 10G1 VL amino acid sequence SEQ ID NO: 24-48: Heavy chain FRs and CDRs from mouse anti-human maturation factor D specific mAb SEQ ID NO: 49-64 Light chain FRs and CDRs from mouse anti-human maturation factor D specific mAb SEQ ID NO: 65-69: CDR consensus sequences derived from mouse anti-human maturation factor D specific mAb SEQ ID NO:70: Human IgG4 constant region SEQ ID NO:71: Human IgG4 constant region with S228P mutation SEQ ID NO:72: Human IgK constant region SEQ ID NO:73:6G6 nucleic acid encoding the HC variable region SEQ ID NO:74:14A11 nucleic acid encoding the HC variable region SEQ ID NO:75:27B3 nucleic acid encoding the HC variable region SEQ ID NO:76:58F5 nucleic acid encoding the HC variable region SEQ ID NO:77:49G3 nucleic acid encoding the HC variable region SEQ ID NO:78:10G1 nucleic acid encoding the HC variable region SEQ ID NO:79:6G6 nucleic acid encoding the LC variable region SEQ ID NO:80:14A11 nucleic acid encoding the LC variable region SEQ ID NO:81:27B3 nucleic acid encoding the LC variable region SEQ ID NO:82:58F5 nucleic acid encoding the LC variable region SEQ ID NO:83:49G3 nucleic acid encoding the LC variable region SEQ ID NO:84:10G1 nucleic acid encoding the LC variable region Anti-human factor D (c-terminal) mAb: VH chain SEQ ID NO:85: mAb clone 3C5 VH amino acid sequence SEQ ID NO:86: mAb clone 11H1 VH amino acid sequence SEQ ID NO:87: mAb clone 12H10 VH amino acid sequence SEQ ID NO:88: mAb clone 7H2 VH amino acid sequence Anti-human factor D (c-terminal) mAb: VL chain SEQ ID NO:89: mAb clone 3C5 VL amino acid sequence SEQ ID NO:90: ​​mAb clone 30H2 VL amino acid sequence SEQ ID NO:91: mAb clone 11H1 VL amino acid sequence SEQ ID NO:92: mAb clone 12H10 VL amino acid sequence SEQ ID NO:93: mAb clone 7H2 VL amino acid sequence SEQ ID NOs: 94-109: Heavy chain FRs and CDRs from mouse anti-human factor D mAbs that bind to epitopes common to mature factor D and pro-factor D SEQ ID NOs: 110-126: Light chain FRs and CDRs from a mouse anti-human factor D mAb that binds to an epitope common to mature factor D and pro-factor D SEQ ID NO: 127:3C5 Nucleic acids encoding the HC and 30H2 variable regions SEQ ID NO:128:11H1 nucleic acid encoding the VH variable region SEQ ID NO:129:12H10 nucleic acid encoding the VH variable region SEQ ID NO:130:7H2 nucleic acid encoding the VH variable region SEQ ID NO:131:3C5 nucleic acid encoding the VL variable region SEQ ID NO:132:30H2 nucleic acid encoding the VL variable region SEQ ID NO:133:11H1 nucleic acid encoding the VL variable region SEQ ID NO:134:12H10 nucleic acid encoding the VL variable region SEQ ID NO:135:7H2 nucleic acid encoding the VL variable region Anti-human profactor D specific mAb: VH chain SEQ ID NO:136: mAb clone 18F5 VH amino acid sequence SEQ ID NO:137: mAb clone 1F9 VH amino acid sequence SEQ ID NO:138: mAb clone 2A4 VH amino acid sequence SEQ ID NO:139: mAb clone 20A1 VH amino acid sequence SEQ ID NO:140: mAb clone 13A10 VH amino acid sequence SEQ ID NO:141: mAb clone 21H1 VH amino acid sequence Anti-human profactor D specific mAb: VL chain SEQ ID NO:142: mAb clone 18F5 VL amino acid sequence SEQ ID NO:143: mAb clone 1F9 VL amino acid sequence SEQ ID NO:144: mAb clone 2A4 VL amino acid sequence SEQ ID NO:145: mAb clone 20A1 VL amino acid sequence SEQ ID NO:146: mAb clone 13A10 VL amino acid sequence SEQ ID NO:147: mAb clone 21H1 VL amino acid sequence SEQ ID NO: 148-174: Heavy chain FRs and CDRs from mouse anti-human profactor D specific mAb SEQ ID NO: 175-200: Light chain FRs and CDRs from mouse anti-human profactor D specific mAb SEQ ID NO: 201-205: CDR consensus sequences derived from mouse anti-human profactor D specific mAb SEQ ID NO:206 nucleic acid encoding the 18F5 HC variable region SEQ ID NO:207 1F9 HC variable region encoding nucleic acid SEQ ID NO:208:2A4 nucleic acid encoding the HC variable region SEQ ID NO:209:20A1 nucleic acid encoding the HC variable region SEQ ID NO:210:13A10 nucleic acid encoding the HC variable region SEQ ID NO:211:21H1 nucleic acid encoding the HC variable region SEQ ID NO:212 nucleic acid encoding the 18F5 LC variable region SEQ ID NO:213 1F9 LC variable region encoding nucleic acid SEQ ID NO:214:2A4 nucleic acid encoding the LC variable region SEQ ID NO:215:20A1 nucleic acid encoding the LC variable region SEQ ID NO:216:13A10 Nucleic acid encoding the LC variable region SEQ ID NO:217:21H1 nucleic acid encoding the LC variable region SEQ ID NO:218: Mouse IgG2a constant region SEQ ID NO:219: Mouse kappa light chain constant region Anti-human MASP-3 inhibitory mAb SEQ ID NO:220:h4D5_VH-14_VH SEQ ID NO:221:h4D5_VL-1-NA SEQ ID NO:222:h4D5_VH-19 SEQ ID NO:223:h10D12_VH-45 SEQ ID NO:224:h10D12_VL-21-GA SEQ ID NO:225:h10D12_VH-49 SEQ ID NO:226:h13B1_VH-9 SEQ ID NO:227:h13B1_VL-1-NA SEQ ID NO:228:h13B1_VH-10 SEQ ID NO:229:h4D5:14_1 NA HC-CDR1 SEQ ID NO:230:h10D12-45-21-GA HC-CDR1 SEQ ID NO:231:h13B1-9-1-NA HC-CDR1 SEQ ID NO:232:h4D5:14_1 NA HC-CDR2 SEQ ID NO:233:h10D12-45-21-GA HC-CDR2 SEQ ID NO:234:h13B1-9-1-NA HC-CDR2 SEQ ID NO:235:h13B1-10-1-NA:HC-CDR2 SEQ ID NO:236:h4D5:14_1 NA HC-CDR3 SEQ ID NO:237:h10D12-45-21-GA HC-CDR3 SEQ ID NO:238:h13B1-9-1-NA HC-CDR3 SEQ ID NO:239:h4D5:14_1 NA LC-CDR1 SEQ ID NO:240:h10D12-45-21-GA LC-CDR1 SEQ ID NO:241:h10D12-45-21-GA LC-CDR2 SEQ ID NO:242:h4D5:14_1 NA LC-CDR3 SEQ ID NO:243:h10D12-45-21-GA LC-CDR3 SEQ ID NO:244:h13B1-9-1-NA LC-CDR3 SEQ ID NO:245: Human IgG4 constant region with S228P and X mutations SEQ ID NO:246: mAb clone 7H2 HC_FR3 amino acid sequence SEQ ID NO:247: mAb clone 2A4 HC_FR1 amino acid sequence

[0025] Detailed Description As described in Examples 1-3, monoclonal antibodies have been generated that specifically bind to the N-terminal region of human mature factor D but not to pro-factor D. As further described in Examples 8-9, monoclonal antibodies have been generated that specifically bind to the propeptide of pro-factor D but not to mature factor D. Mature factor D-specific monoclonal antibodies and pro-factor D-specific antibodies are useful for detecting the mature and / or pro-forms of factor D in biological samples and may be used to determine the status of the alternative pathway of complement (APC) in a mammalian subject. As further described in Examples 10-12, mature factor D-specific monoclonal antibodies may also be used to determine the status of factor D after treatment with a MASP-3 inhibitor that inhibits the conversion of pro-factor D to mature factor D. Thus, in one aspect, the present invention is directed to monoclonal antibodies that specifically bind to the N-terminal region of human mature factor D and the use of these antibodies in methods for detecting the presence or amount of mature factor D in a biological sample. In another aspect, the present invention is directed to monoclonal antibodies that specifically bind to the activation (pro)peptide of pro-factor D and the use of these antibodies in methods for detecting the presence or amount of pro-factor D in a biological sample. In another aspect, the present invention is directed to the use of mature factor D-specific monoclonal antibodies and / or pro-factor D-specific monoclonal antibodies to measure the presence or amount of mature factor D and / or pro-factor D in a mammalian subject before and after treatment with a MASP-3 inhibitor, such as a high-affinity MASP-3 inhibitory antibody, wherein the MASP-3 inhibitory antibody is capable of inhibiting the conversion of pro-factor D to mature factor D, thereby inhibiting APC.

[0026] I. Definition Unless otherwise defined herein, all terms used herein have the same meaning as would be understood by one of ordinary skill in the art to which the invention pertains. The following definitions are provided for the purpose of providing clarity regarding terms used in the specification and claims to describe the invention.

[0027] The terms "antibody" and "immunoglobulin" are used interchangeably herein. These terms are well understood by those skilled in the art and refer to a protein consisting of one or more polypeptides that specifically bind to an antigen. One form of antibody constitutes the basic structural unit of an antibody. This form is a tetramer, consisting of two identical pairs of antibody chains, each pair having one light chain and one heavy chain. In each pair, the variable regions of the light and heavy chains together are responsible for binding to an antigen, and the constant regions are responsible for antibody effector functions.

[0028] As used herein, the term "antibody" includes antibodies and antibody fragments thereof derived from any antibody-producing mammal (e.g., mouse, rat, rabbit, and primate, including human), or derived from hybridoma, phage selection, recombinant expression, or transgenic animals (or other methods of producing antibodies or antibody fragments), that specifically bind to an antigen, e.g., human pro-factor D shown as SEQ ID NO:2 (e.g., an epitope in the propeptide "APPRGR" shown as SEQ ID NO:4), or human mature factor D shown as SEQ ID NO:3 (e.g., an epitope at the N-terminus of mature factor D comprising or consisting of "ILGGREA" shown as SEQ ID NO:5), or that binds an epitope common to human pro-factor D and human mature factor D (e.g., an epitope in the C-terminal region of factor D (e.g., amino acids 8-228 of SEQ ID NO:3)). The term "antibody" is not intended to be limiting with regard to the source of the antibody or the manner in which it is made (e.g., by hybridoma, phage selection, recombinant expression, transgenic animals, peptide synthesis, etc.). Exemplary antibodies include polyclonal, monoclonal, and recombinant antibodies; multispecific antibodies (e.g., bispecific antibodies); humanized antibodies; fully human antibodies, murine antibodies; chimeric, mouse-human, mouse-primate, primate-human monoclonal antibodies; and anti-idiotypic antibodies, and may be any intact molecule or fragment thereof. As used herein, the term "antibody" encompasses not only intact polyclonal or monoclonal antibodies, but also fragments thereof (e.g., dAb, Fab, Fab', F(ab')2, Fv), single chain (ScFv), synthetic variants thereof, naturally occurring variants, fusion proteins comprising an antibody portion having an antigen-binding fragment with the required specificity, humanized antibodies, chimeric antibodies, and any other modified conformation of an immunoglobulin molecule containing an antigen-binding site or fragment (epitope recognition site) with the required specificity.

[0029] As used herein, the term "antigen-binding fragment" refers to a polypeptide fragment containing at least one CDR of an immunoglobulin heavy chain and / or light chain that specifically binds to an antigen, e.g., human pro-factor D shown as SEQ ID NO:2 (e.g., an epitope in the propeptide "APPRGR" shown as SEQ ID NO:4), or human mature factor D shown as SEQ ID NO:3 (e.g., an epitope at the N-terminus of mature factor D comprising or consisting of "ILGGREA" shown as SEQ ID NO:5), or an epitope common to human pro-factor D and human mature factor D (e.g., an epitope in the C-terminal region of factor D (e.g., amino acids 8-228 of SEQ ID NO:3)). In this regard, an antigen-binding fragment of an antibody described herein can include one, two, three, four, five, or all six of the CDRs of the VH and VL sequences, e.g., one, two, three, four, five, or six CRSs of the VH and VL sequences from the disclosed anti-human factor D antibodies shown herein.

[0030] As used herein, the term "anti-factor D monoclonal antibody" refers to a homogenous antibody population in which the monoclonal antibody is composed of amino acids that are involved in selective binding of an epitope on human factor D, for example, human pro-factor D shown as SEQ ID NO:2 (e.g., an epitope in the propeptide "APPRGR" shown as SEQ ID NO:4), or that specifically binds to human mature factor D shown as SEQ ID NO:3 (e.g., an epitope at the N-terminus of mature factor D comprising or consisting of "ILGGREA" shown as SEQ ID NO:5), or that binds to an epitope common to human pro-factor D and human mature factor D (e.g., an epitope in the C-terminal region of factor D (e.g., amino acids 8-228 of SEQ ID NO:3)). The term "monoclonal antibody" encompasses not only intact and full-length monoclonal antibodies, but also fragments thereof (e.g., Fab, Fab', F(ab')2, Fv), single chain (ScFv), variants thereof, fusion proteins comprising an antigen-binding portion, humanized monoclonal antibodies, chimeric monoclonal antibodies, and any other modified conformation of an immunoglobulin molecule containing an antigen-binding fragment (epitope recognition site) with the required specificity and epitope-binding ability.

[0031] As used herein, the modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not intended to be limiting as regards the source of the antibody or the manner in which it is made (e.g., by hybridoma, phage selection, recombinant expression, transgenic animals, etc.). The term includes both whole immunoglobulins and fragments thereof, as discussed above in the definition of "antibody." Monoclonal antibodies may be obtained using any technique that provides for the production of antibody molecules by continuous cell line culture, such as the hybridoma method described by Kohler, G., et al., Nature 256:495, 1975, or they may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567 to Cabilly). Monoclonal antibodies may also be isolated from phage antibody libraries using the techniques described in Clackson, T., et al., Nature 352:624-628, 1991, and Marks, JD, et al., J. Mol. Biol. 222:581-597, 1991. Such antibodies can be of any immunoglobulin class, including IgG, IgM, IgE, IgA, IgD, and any subclass thereof.

[0032] Recognized immunoglobulin polypeptides include kappa and lambda light chains and alpha, gamma (IgG1, IgG2, IgG3, IgG4), delta, epsilon, and mu heavy chains, or equivalents in other species. Full-length immunoglobulin "light chains" (about 25 kDa or about 214 amino acids) contain a variable region of about 110 amino acids at the NH2-terminus and a kappa or lambda constant region at the COOH-terminus. Full-length immunoglobulin "heavy chains" (about 50 kDa or about 446 amino acids) similarly contain a variable region (of about 116 amino acids) and one of the aforementioned heavy chain constant regions, e.g., gamma (of about 330 amino acids).

[0033] The basic four-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. IgM antibodies consist of five basic heterotetrameric units plus an additional polypeptide called the J chain, and therefore contain 10 antigen-binding sites. Secreted IgA antibodies can polymerize to form multivalent assemblies containing two to five basic four-chain units plus the J chain. Each L chain is linked to an H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds, depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. The VH and VL pair together to form a single antigen-binding site.

[0034] Each H chain has at its N-terminus a variable domain (VH) followed by three constant domains (CH) for each of the α and γ chains and four CH domains (CH) for the μ and ε isotypes.

[0035] Each L chain has a variable domain (VL) at its N-terminus followed by a constant domain (CL) at its other end. The VL is aligned with the VH, and the CL is aligned with the first constant domain (CH1) of the heavy chain. L chains from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (κ) and lambda (λ), based on the amino acid sequence of their constant domain (CL).

[0036] Depending on the amino acid sequence of the constant domain (CH) of their heavy chains, immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), respectively. The γ and α classes are further divided into subclasses based on slight differences in CH sequence and function; for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0037] For the structure and properties of various classes of antibodies, see, e.g., Basic and Clinical Immunology, 8th Edition, Daniel P. Stites, Abba I. Terr and Tristram G. Parslow (eds); Appleton and Lange, Norwalk, Conn., 1994, page 71 and Chapter 6.

[0038] The term "variable" refers to the fact that certain segments of V domains vary widely in sequence among antibodies. V domains mediate antigen binding and define the specificity of a particular antibody for its particular antigen. However, variability is not evenly distributed across the 110 amino acid length of the variable domains. Rather, V regions consist of relatively invariant stretches of 15-30 amino acids called framework regions (FRs), separated by shorter regions of extreme variability called "hypervariable regions," each 9-12 amino acids long. Naturally occurring heavy and light chain variable domains each contain four FRs in a largely beta-sheet configuration connected by three hypervariable regions (which form loops connecting, and in some cases, part of, the n-sheet structure). The hypervariable regions in each chain are held in close proximity to the FRs and together with the hypervariable regions from the other chains, contribute to the formation of the antigen-binding site of antibodies (see Kabat, et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The constant domains are not directly involved in antibody binding to antigens, but exhibit various effector functions, such as antibody-dependent cellular cytotoxicity (ADCC).

[0039] As used herein, the term "hypervariable region" refers to the amino acid residues of an antibody responsible for antigen binding. Hypervariable regions generally consist of amino acid residues from the "complementarity-determining regions" or "CDRs" (i.e., residues approximately 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain, and approximately 31-35 (H1), 50-66 (H2), and 95-102 (H3) in the heavy chain variable domain, as numbered according to the Kabat numbering system as described in Kabat, et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)); and / or residues from the "hypervariable loops" (i.e., residues approximately 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain, and approximately 31-35 (H1), 50-66 (H2), and 95-102 (H3) in the heavy chain variable domain, as numbered according to the Kabat numbering system as described in Chothia and Lesk, J. Mol. Biol. 196:901-917). residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain, and residues 26-32 (H1), 52-56 (H2), and 95-101 (H3) in the heavy chain variable domain, when numbered according to the Chothia numbering system as described in (1987); and / or residues from the "hypervariable loops" / CDRs (e.g., Lefranc, J.P., et al., Nucleic Acids Res 27:209-212; Ruiz, M., et al., Nucleic Acids Res 28:219-221). (2000) and includes residues 27-38 (L1), 56-65 (L2), and 105-120 (L3) in VL, and 27-38 (H1), 56-65 (H2), and 105-120 (H3) in VH.

[0040] As used herein, the term "antibody fragment" refers to a portion derived from or related to a full-length anti-factor D antibody, generally comprising the antigen-binding or variable region thereof. Illustrative examples of antibody fragments include Fab, Fab', F(ab)2, F(ab')2, and Fv fragments, scFv fragments, diabodies, linear antibodies, single-chain antibody molecules, bispecific and multispecific antibodies formed from antibody fragments.

[0041] As used herein, a "single-chain Fv" or "scFv" antibody fragment refers to a single-chain Fv fragment of an antibody. H and V L Fv polypeptides generally comprise V domains, and these domains are present in a single polypeptide chain. H Domains and V L The scFv further comprises a polypeptide linker between the domains, which enables the scFv to form the desired structure for antigen binding. See Pluckthun in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994). An "Fv" is the minimum antibody fragment containing a complete antigen recognition and binding site. This fragment consists of a dimer of one heavy-chain variable region domain and one light-chain variable region domain in tight, non-covalent association. The folding of these two domains results in six hypervariable loops (three loops each from the H and L chains) that provide amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific CDRs) has the ability to recognize and bind antigen, although with lower affinity than the entire binding site.

[0042] As used herein, a "humanized antibody" is a chimeric molecule, generally prepared using recombinant techniques, that has an antigen-binding site derived from an immunoglobulin from a non-human species and the remaining immunoglobulin structure of the molecule based on the structure and / or sequence of a human immunoglobulin. The antigen-binding site can comprise either a complete variable region fused onto a constant domain or only CDRs grafted onto appropriate framework regions within the variable domain. The epitope-binding site can be wild-type or modified by one or more amino acid substitutions. Another approach focuses not only on providing human-derived constant regions, but also on modifying the variable regions to reshape them as closely as possible to human form. In some embodiments, a humanized antibody preserves all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs from a mouse antibody). In other embodiments, a humanized antibody has one or more (one, two, three, four, five, six) CDRs that are altered relative to the original antibody, also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody.

[0043] As used herein, the term "specific binding" refers to the ability of an antibody to preferentially bind to a particular analyte present in a homogeneous mixture of various analytes. In certain embodiments, the specific binding interaction distinguishes between desired and undesired analytes in a sample, and in some embodiments, by about 10-100 fold or more (e.g., about 1000 fold or more than 10,000 fold). In certain embodiments, the affinity of the capture agent and analyte when specifically bound in a capture agent / analyte complex is less than about 100 nM, or less than about 50 nM, or less than about 25 nM, or less than about 10 nM, or less than about 5 nM, or less than about 1 nM. D It is characterized by a dissociation constant.

[0044] As used herein, the term "variant" antibody refers to a molecule whose amino acid sequence differs from that of a "parent" or reference antibody amino acid sequence by the addition, deletion, and / or substitution of one or more amino acid residues in the parent antibody sequence. In one embodiment, a variant anti-factor D antibody refers to a molecule containing variable regions that are identical to the parent variable domains except for a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in the CDR regions of the heavy chain variable region, and / or a total of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in the CDR regions of the light chain variable region. In some embodiments, the amino acid substitutions are conservative sequence modifications.

[0045] As used herein, the term "parent antibody" refers to an antibody encoded by an amino acid sequence used to prepare a variant. Preferably, the parent antibody has human framework regions and, if present, human antibody constant regions. For example, the parent antibody can be a humanized antibody or a fully human antibody.

[0046] As used herein, the term "isolated antibody" refers to an antibody that has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of its natural environment are substances that may interfere with the antibody's diagnostic or therapeutic use, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In preferred embodiments, the antibody is purified (1) to greater than 95% by weight, and most preferably greater than 99% by weight, of the antibody as determined by the Lowry method; (2) sufficiently to obtain at least 15 residues of N-terminal or internal amino acid sequence using a spinning cup sequencer; or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibodies include antibodies in situ within recombinant cells, since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibodies will be prepared by at least one purification step.

[0047] As used herein, the term "epitope" refers to a portion of an antigen to which a monoclonal antibody specifically binds. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three-dimensional structural characteristics, as well as specific charge characteristics. More specifically, the term "pro-factor D epitope" as used herein refers to a portion of the corresponding polypeptide (SEQ ID NO:4) to which an antibody immunospecifically binds as determined by any method known in the art, e.g., by immunoassay. The term "mature factor D epitope" as used herein refers to an epitope encompassing the amino-terminal portion of the corresponding polypeptide (SEQ ID NO:3) to which an antibody immunospecifically binds as determined by any method known in the art, e.g., by immunoassay, e.g., an epitope at the N-terminus of mature factor D comprising or consisting of "ILGGREA" as shown as SEQ ID NO:5. The term "epitope common to pro-factor D and mature factor D," as used herein, refers to an epitope in the C-terminal region common to pro-factor D and mature factor D (e.g., amino acids 8-228 of SEQ ID NO:3) to which an antibody immunospecifically binds as determined by any method known in the art, e.g., by immunoassay. An antigenic epitope need not necessarily be immunogenic. Such epitopes may be linear in nature or may be discontinuous epitopes. Thus, as used herein, the term "conformational epitope" refers to a discontinuous epitope formed by a spatial relationship between amino acids of an antigen other than a contiguous series of amino acids.

[0048] As used herein, a "mammalian subject" includes, but is not limited to, humans, non-human primates, dogs, cats, horses, sheep, goats, cattle, rabbits, pigs, and rodents.

[0049] As used herein, the term "biological sample" includes, but is not limited to, blood, plasma, serum, sputum, amniotic fluid, cerebrospinal fluid, cell lysate, peritoneal fluid, urine, saliva, and tissue.

[0050] As used herein, the term "contacting" refers to the combined act of bringing an antibody of the present invention and a biological sample into contact such that a binding interaction occurs between the antibody and a target protein (e.g., pro-factor D or mature factor D) in the biological sample.

[0051] As used herein, the term "detecting antibody" or "detection antibody" refers to an antibody that enables detection. The detection antibody can be directly or indirectly (e.g., through another antibody) conjugated to a detectable label or signal or to a signal-generating moiety. The signal can be a radioactive signal (e.g., radioactive iodine, tritium, carbon, sulfur, etc.), a chromogenic signal, a fluorescent signal, etc. Signal-generating moieties that act on a signal-generating substrate include, but are not limited to, horseradish peroxidase (HRP) [suitable substrates include 3,3',5,5'-tetramethylbenzidine (TMB); OPD; 2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid diammonium salt]; alkaline phosphatase [suitable substrates include p-nitrophenyl phosphate disodium salt]; and beta-galactosidase [suitable substrates include O-nitrophenyl-beta-D-galactopyranoside]. The signal can be amplified by using an avidin-biotin conjugation system. Detectable labels or signal-generating moieties can be directly and / or indirectly coupled to the anti-factor D antibodies and antigen-binding fragments thereof of the present invention. For example, immunoconjugates can include anti-factor D antibodies labeled with radioisotopes or enzyme activity that allow detection in immunoassays.

[0052] As used herein, the term "MASP-3 inhibitor" refers to any agent, including anti-MASP-3 antibodies and their MASP-3-binding fragments, natural and synthetic peptides, competitive substrates, small molecules, and expression inhibitors, that bind to MASP-3 and inhibit the conversion of pro-factor D to mature factor D, thereby inhibiting alternative pathway activation of complement (APC). Exemplary MASP-3 inhibitory antibodies are disclosed in WO2018 / 026722, which is incorporated herein by reference. In some embodiments, the MASP-3 inhibitor is a MASP-3 inhibitory antibody, for example, a MASP-3 inhibitory monoclonal antibody selected from the group consisting of 4D5, 10D12, and 13B1.

[0053] As used herein, amino acid residues are abbreviated as follows: alanine (Ala; A), asparagine (Asn; N), aspartic acid (Asp; D), arginine (Arg; R), cysteine ​​(Cys; C), glutamic acid (Glu; E), glutamine (Gln; Q), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V).

[0054] In the broadest sense, naturally occurring amino acids can be divided into groups based on the chemical characteristics of their side chains. A "hydrophobic" amino acid refers to any of Ile, Leu, Met, Phe, Trp, Tyr, Val, Ala, Cys, or Pro. A "hydrophilic" amino acid refers to any of Gly, Asn, Gln, Ser, Thr, Asp, Glu, Lys, Arg, or His. This grouping of amino acids can be further subclassified as follows: An "uncharged hydrophilic" amino acid refers to any of Ser, Thr, Asn, or Gln. An "acidic" amino acid refers to any of Glu or Asp. A "basic" amino acid refers to any of Lys, Arg, or His.

[0055] As used herein, the term "conservative amino acid substitutions" is exemplified by substitutions between amino acids within each of the following groups: (1) glycine, alanine, valine, leucine, and isoleucine, (2) phenylalanine, tyrosine, and tryptophan, (3) serine and threonine, (4) aspartic acid and glutamic acid, (5) glutamine and asparagine, and (6) lysine, arginine, and histidine.

[0056] As used herein, an "isolated nucleic acid molecule" refers to a nucleic acid molecule (e.g., a polynucleotide) that is not integrated into the genomic DNA of an organism. For example, a DNA molecule encoding a growth factor that is separated from the genomic DNA of a cell is an isolated DNA molecule. Another example of an isolated nucleic acid molecule is a chemically synthesized nucleic acid molecule that is not integrated into the genome of an organism. A nucleic acid molecule isolated from a particular species is smaller than the complete DNA molecule of a chromosome from that species.

[0057] As used herein, a "nucleic acid molecule construct" is a nucleic acid molecule, either single-stranded or double-stranded, that has been modified through human intervention to contain segments of nucleic acid combined and arranged in an arrangement that does not occur in nature.

[0058] As used herein, an "expression vector" is a nucleic acid molecule that encodes a gene to be expressed in a host cell. Typically, an expression vector comprises a transcription promoter, a gene, and a transcription terminator. Gene expression is usually placed under the control of a promoter, and such a gene is said to be "operably linked to" that promoter. Similarly, if a regulatory element modulates the activity of a core promoter, that regulatory element and that core promoter are operably linked.

[0059] As used herein, the term "approximately" or "about" in reference to a numerical value is generally deemed to include numerical values ​​within a range of 5% in either direction (greater or less) of that numerical value, unless otherwise stated or clear from the context (except in cases where such numerical value may exceed 100% of the possible value). When a range is stated, the endpoints are included within the range unless otherwise stated or clear from the context.

[0060] As used herein, the singular forms "a," "an," and "the" include plural aspects unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes not only a single cell but also two or more cells; reference to "an agent" includes not only one agent but also two or more agents; reference to "an antibody" includes a plurality of such antibodies, reference to "a framework region" includes reference to one or more framework regions and equivalents thereof known to those skilled in the art, and so forth.

[0061] Amino acid sequence identity percentage (%) is defined as the percentage of amino acids in the candidate sequence that are identical to the amino acids in the reference sequence, after aligning the sequences to achieve maximum sequence identity percentage, and if necessary, introducing gaps.The alignment for determining sequence identity percentage can be achieved by various methods within the skill of the art, for example, by using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software.Appropriate parameters for measuring alignment can be determined by known methods, including any algorithm required to achieve maximum alignment over the entire length of the sequence being compared.

[0062] Each aspect herein should apply to all other aspects mutatis mutandis unless expressly stated otherwise.

[0063] Standard techniques may be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection). Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein. These and related techniques and procedures may generally be performed according to conventional methods well known in the art and as described in the various general and more specific references cited and discussed throughout this specification. See, for example, Sambrook et al., 2001, MOLECULAR CLONING: A LABORATORY MANUAL, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (Greene Publ. Assoc. Inc. & John Wiley & Sons, Inc., NY, NY); Current Protocols in Immunology (Edited by: John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strober 2001 John Wiley & Sons, NY, NY); or other relevant Current Protocol publications and other similar references. Unless specific definitions are provided, the nomenclature used in connection with, and the laboratory procedures and techniques of, molecular biology, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for recombinant technology, molecular biological synthesis, microbiological synthesis, chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0064] It is contemplated that any embodiment discussed herein can be implemented with respect to any method, kit, reagent, or composition of the invention, and vice versa. Furthermore, the compositions of the invention can also be used to practice the methods of the invention.

[0065] II. Overview As described in Examples 1-3 herein, the present invention provides monoclonal anti-factor D antibodies (also referred to as mature factor D-specific antibodies) that specifically bind to human mature factor D but not to pro-factor D. As described in Examples 8-9, the present invention also provides monoclonal anti-factor D antibodies (also referred to as pro-factor D-specific antibodies) that specifically bind to pro-factor D but not to mature factor D. As described in Examples 4-5 herein, the present invention also provides monoclonal anti-factor D antibodies that bind to both pro- and mature factor D. As described in Examples 6 and 7, mature factor D-specific monoclonal antibodies and pro-factor D-specific monoclonal antibodies are useful for detecting the mature and / or pro-forms of factor D in biological samples and may be used to determine the status of the alternative pathway of complement (APC) in a mammalian subject. As further described in Examples 10-12, mature factor D-specific monoclonal antibodies and / or pro-factor D-specific antibodies may also be used to determine the status of factor D after treatment with a MASP-3 inhibitor, such as a MASP-3 inhibitory antibody that inhibits the conversion of pro-factor D to mature factor D.

[0066] Thus, in one aspect, the present invention is directed to monoclonal antibodies that specifically bind to the N-terminal region of human maturation factor D and the use of these antibodies in methods for detecting the presence or amount of mature factor D in a biological sample. In another aspect, the present invention is directed to monoclonal antibodies that specifically bind to the activation (pro)peptide of pro-factor D and the use of these antibodies in methods for detecting the presence or amount of pro-factor D in a biological sample. In another aspect, the present invention is directed to the use of a mature factor D-specific monoclonal antibody and / or a pro-factor D-specific monoclonal antibody to measure the presence or amount of mature factor D and / or pro-factor D in a mammalian subject before and after treatment with a MASP-3 inhibitor, such as a high-affinity MASP-3 inhibitory antibody, wherein the MASP-3 inhibitory antibody is capable of inhibiting the conversion of pro-factor D to mature factor D, thereby inhibiting APC.

[0067] Therefore, the subject antibodies can be used in diagnostic methods to detect the presence or amount of mature factor D and / or pro-factor D in a biological sample obtained from a subject. In one embodiment, the presence or amount of mature factor D and / or pro-factor D is useful as a biomarker for determining the effectiveness of a MASP-3 inhibitor in inhibiting APC and / or for monitoring the dosing of a subject undergoing treatment with a MASP-3 inhibitor, such as a MASP-3 inhibitory antibody (e.g., MASP-3 inhibitory antibody 4D5, 10D12 or 13B1).

[0068] III. Overview of the Complement System Figure 1 illustrates three pathways that drive complement activation in response to distinct initiating events: the classical pathway, the lectin pathway, and the alternative pathway (Noris M., Semin Nephrol 33 (6):479-92, 2013). The classical pathway is triggered by immune complexes and mediates important immune effector functions through the early activation of two proteases, C1r and C1s. The lectin pathway can be activated by specific types of cell surface glycan patterns that are typically found on microorganisms or damaged host tissue but not on healthy host cells. Lectin pathway activation is initiated by a group of enzymes known as mannan-binding lectin-associated serine proteases-1 and 2 (MASP-1 and MASP-2) (Yongqing T. et al., Biochim Biophys Acta 1824 (1):253-62, 2012). These proteases form complexes with lectins, such as mannan-binding lectin (MBL), ficolins, and collectins 10 and 11. Lectins bind to carbohydrate patterns on foreign or damaged host cells, thereby targeting the proteolytic activity of MASPs to specific surfaces. Complement factor D (CFD) is a serine protease essential for activation of the alternative pathway. As shown in Figure 1, factor D (CFD) is expressed as an inactive zymogen (referred to herein as "pro-factor D"), and CFD circulates primarily in plasma as a cleaved mature serine protease (referred to herein as "mature factor D"). As described in WO2013 / 180834 and WO2013 / 192240, it has recently been determined that MASP-3 is responsible for converting complement factor D (CFD) from its proenzyme form (pro-factor D) to its mature form (mature factor D), and thus the MASP-3 protein is in an important upstream regulatory position for the alternative pathway. As further described in WO2018 / 026722, which is incorporated herein by reference, a number of high-affinity anti-MASP-3 inhibitory antibodies have been generated that bind to the serine protease domain of MASP-3 and inhibit its catalytic activity, thereby inhibiting the conversion of pro-factor D to mature factor D and blocking activation of the alternative pathway.

[0069] The primary function of the complement system, part of the innate immune response, is to protect the host from infectious agents (Ricklin et al., Nat Immunol 11 (9):785-97, 2010). Through the coordinated action of protein complex assembly and proteolytic cascades, this complex physiological system directs immune and inflammatory responses to surfaces displaying molecular patterns not normally present on healthy host cells. Complement system activation ultimately leads to target cell destruction, either through the formation of membrane attack complexes (MACs), which directly disrupt pathogen membranes, causing cell lysis, or through opsonization, which facilitates the uptake of infectious agents by phagocytes, as shown in Figure 1. In addition, substrate cleavage by complement proteases releases cytokine-like peptides called anaphylatoxins, which induce several important biological activities, such as leukocyte recruitment and immune cell activation.

[0070] The alternative complement pathway (APC) is generally described as a downstream amplifier of complement activity, enhancing the host immune response following complement activation via the classical and lectin pathways. However, the APC's ability to generate a positive feedback loop of protease complexes that actively drive the formation of new complexes of the same type is unique within the complement pathway (Lachmann et al., Adv Immunol 104:115-49, 2009). The APC C3 convertase, a self-propagating complex, is composed of two proteins: C3b and Bb. The newly formed C3b can covalently attach to local surfaces via a thioester bond and function as a potent opsonin, targeting marked cells for phagocytosis and destruction. Additionally, C3b also provides a scaffold for the binding and activation of complement factor B (CFB) (Lachmann et al., Adv Immunol 104:115-49, 2009; Norris et al., Semin Nephrol 33 (6):479-92, 2013). In complex with C3b, CFB adopts the appropriate conformation for cleavage by complement factor D (CFD). This cleavage event converts the single-chain polypeptide into a non-catalytic fragment (Ba) and a catalytic fragment (Bb). The Ba fragment is released from the complex; however, the Bb fragment remains associated with C3b to generate the active APC C3 convertase C3bBb (Lachmann et al., Adv Immunol 104:115-49, 2009; Norris et al., Semin Nephrol 33 (6):479-92, 2013). The ability of C3bBb to cleave additional C3 and generate multiple new convertases provides a robust signal amplification mechanism.

[0071] While the complement system supports innate host defense against pathogens, dysregulated and unabated complement activity can also function as a major driver of disease, leading to uncontrolled inflammation and the spread of tissue destruction. In many situations, the APC and C3b amplification loop play a critical role in determining the magnitude of the complement response and its downstream consequences. Thus, therapeutic modulation of APC by inhibiting Bb activity or blocking CFB activation through cleavage by CFD is a well-characterized potential control point for treating many APC-mediated autoimmune and inflammatory diseases.

[0072] As mentioned above, MASP-3 is responsible for converting CFD from its proenzyme form to its mature form, demonstrating that the MASP-3 protein plays an important upstream regulatory role in the alternative pathway. In wild-type animal or human plasma, the majority of systemic CFD has already been processed to its mature form by in vivo MASP-3 activity, making in vitro evaluation of APC inhibition by MASP-3 inhibitors impossible using conventional assays. Therefore, there is a need for detection reagents and assays for measuring the presence and amount of pro-factor D and / or mature factor D in biological samples that can be used as biomarkers of APC status and can also be used to evaluate APC inhibition by MASP-3 inhibitors in vitro and / or in vivo.

[0073] IV. Anti-factor D antibodies As mentioned above, complement factor D (CFD) is a serine protease essential for APC activation. As shown in Figure 1, factor D (CFD) is expressed as an inactive zymogen (referred to herein as "pro-factor D"), and CFD circulates in plasma primarily as a cleaved mature serine protease (referred to herein as "mature factor D").

[0074] Figure 2 provides the amino acid sequences of (i) human full-length Factor D (SEQ ID NO:1), including the signal sequence aa 1-19 (shown in italics) and the underlined activation (pro)peptide; (ii) human pro-Factor D (SEQ ID NO:2), with the underlined propeptide; and (iii) human mature Factor D (SEQ ID NO:3). As shown in Figure 2, the propeptide of human pro-Factor D is ("APPRGR" (SEQ ID NO:4).

[0075] Figure 3 provides an alignment of the amino acid sequences of complement factor D (full length) from various species, including Homo sapiens (SEQ ID NO:1); Macaque (SEQ ID NO:8); Canis (SEQ ID NO:9); Rattus (SEQ ID NO:10); and Mus musculus (SEQ ID NO:11). The italicized portion of each sequence depicts the signal sequence, and the underlined portion depicts the activation "pro" peptide sequence.

[0076] As shown in Figures 2 and 3, the factor D protein comprises an N-terminal pro-region, an activation peptide region, and the remaining C-terminal region. Mature factor D has a unique amino terminus compared to pro-factor D in each species (e.g., the N-terminus begins at residue 26 of human full-length factor D (SEQ ID NO:1)). Mature factor D and pro-factor D share a common sequence in the C-terminal portion of their respective proteins (e.g., amino acids 27-253 of SEQ ID NO:1).

[0077] A. Anti-human maturation factor D specific monoclonal antibody As described in Examples 1 and 2 herein, the inventors used the peptide "ILGGREA" (SEQ ID NO:5), corresponding to amino acid residues 1-7 of the amino-terminal region of human maturation factor D (SEQ ID NO:3), as an antigen to generate anti-maturation factor D-specific antibodies suitable for use in the detection assays and methods described herein. As shown in Figure 3, the N-terminal sequence "ILLGGREA" (SEQ ID NO:5) is conserved between human (Homo sapiens) and macaque (Macaca) maturation factor D proteins.

[0078] As described in Example 2, the variable heavy and light chain fragments of several representative anti-mature factor D specific monoclonal antibodies were cloned and sequenced.

[0079] Figure 7A is an amino acid sequence alignment of the variable heavy chain regions of six anti-mature factor D-specific clones identified as having high binding affinity to the N-terminal peptide of mature factor D, "ILGGREA" (SEQ ID NO:5).

[0080] Figure 7B is an amino acid sequence alignment of the variable light chain regions of six anti-mature factor D-specific clones identified as having high binding affinity to the N-terminal peptide of mature factor D, "ILGGREA" (SEQ ID NO:5).

[0081] The heavy and light chain variable regions and CDRs therein of six mature Factor D-specific antibodies are provided in Tables 1 and 2 below.

[0082] Table 1: Anti-human maturation factor D specific antibody sequences: Mouse parent TIFF2025076475000033.tif69162

[0083] Table 2: Anti-human maturation factor D specific antibodies: CDRs TIFF2025076475000034.tif63160

[0084] Thus, in one aspect, the present invention provides an isolated antibody, or antigen-binding fragment thereof, that specifically binds to an epitope in the amino-terminal (N-terminal) region of human maturation factor D, the epitope comprising or consisting of the amino acid sequence "ILGGREA" (SEQ ID NO:5). In one embodiment, the maturation factor D-specific antibody or fragment thereof specifically binds to human maturation factor D (SEQ ID NO:3) and does not bind to human pro-factor D (SEQ ID NO:2). In one embodiment, the maturation factor D-specific antibody is a monoclonal antibody. In one embodiment, the maturation factor D-specific antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. In one embodiment, the maturation factor D-specific antibody fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. In one embodiment, the maturation factor D-specific antibody is a single-chain molecule. In one embodiment, the maturation factor D-specific antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. In one embodiment, the mature factor D-specific antibody or antigen-binding fragment thereof has a K of less than 10 nM. D In one embodiment, the mature factor D-specific antibody or antigen-binding fragment thereof is labeled with a detectable moiety, e.g., a detectable moiety suitable for use in an immunoassay as further described herein. In one embodiment, the mature factor D-specific antibody or fragment thereof is immobilized on a substrate, e.g., a substrate suitable for use in an immunoassay as further described herein.

[0085] In one embodiment, a maturation factor D-specific antibody or fragment thereof (i.e., an antibody or fragment thereof that specifically binds to human maturation factor D) comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12-17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18-23, wherein the CDRs are numbered according to the Kabat numbering system. In one embodiment, a maturation factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO:65), wherein X at position 1 is T, I, or S, and X at position 3 is G or I; (b) amino acid sequence HC-CDR2 containing TIFF2025076475000035.tif4128 (in the sequence, X at position 11 is H or N, and X at position 16 is S or R); (c) amino acid sequence HC-CDR3 containing TIFF2025076475000036.tif4128 (in which X at position 6 is R, G, or N, X at position 7 is S or Y, X at position 8 is F, I, or V, and X at position 10 is D or H); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69) (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54).

[0086] In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:25, (b) HC-CDR2 comprising SEQ ID NO:27, (c) HC-CDR3 comprising SEQ ID NO:29, (d) LC-CDR1 comprising SEQ ID NO:50, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:12 or SEQ ID NO:13. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:19. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO:12 and a VL comprising SEQ ID NO:18. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO:13 and a VL comprising SEQ ID NO:19.

[0087] In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:33, (b) HC-CDR2 comprising SEQ ID NO:34, (c) HC-CDR3 comprising SEQ ID NO:36, (d) LC-CDR1 comprising SEQ ID NO:58, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:14. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 20. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 14, and a VL comprising SEQ ID NO: 20.

[0088] In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:38, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:41, (d) LC-CDR1 comprising SEQ ID NO:60, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:15. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 21. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 15, and a VL comprising SEQ ID NO: 21.

[0089] In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:43, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:41, (d) LC-CDR1 comprising SEQ ID NO:62, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:16. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 22. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 22.

[0090] In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:43, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:47, (d) LC-CDR1 comprising SEQ ID NO:63, (e) LC-CDR2 comprising SEQ ID NO:64, and (f) LC-CDR3 comprising SEQ ID NO:54. In one embodiment, the mature factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:17. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 23. In one embodiment, the mature Factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 17, and a VL comprising SEQ ID NO: 23.

[0091] In certain embodiments, a maturation factor D-specific antibody or fragment thereof that specifically binds to human maturation factor D has a heavy chain variable domain that is substantially identical (e.g., at least about 70%, at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least 99% identical) to any of the heavy chain variable domain sequences shown in Table 1. In certain embodiments, a maturation factor D-specific antibody or fragment thereof that specifically binds to human maturation factor D has a light chain variable domain that is substantially identical (e.g., at least about 70%, at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least 99% identical) to any of the light chain variable domain sequences shown in Table 1.

[0092] In another aspect, the present disclosure provides a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy chain variable region of a maturation factor D-specific antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NOs: 12-17, and the CDRs are numbered according to the Kabat numbering system. In another aspect, the present disclosure provides a nucleic acid encoding the complementarity determining regions (CDRs) of the light chain variable region of a maturation factor D-specific antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NOs: 18-23, and the CDRs are numbered according to the Kabat numbering system.

[0093] In another aspect, the disclosure provides a cloning or expression vector comprising a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 12-17, the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 18-23, and the CDRs are numbered according to the Kabat numbering system.

[0094] In another aspect, the disclosure provides a cell containing a cloning or expression vector comprising a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 12-17, the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 18-23, and the CDRs are numbered according to the Kabat numbering system.

[0095] In another aspect, the present disclosure provides a method for producing a human maturation factor D-specific antibody, comprising culturing cells containing an expression vector containing a nucleic acid encoding one or both of the heavy and light chain polypeptides of any of the maturation factor D-specific antibodies or antigen-binding fragments disclosed herein. The cell or culture of cells is cultured under conditions and for a time sufficient to allow the cell (or culture of cells) to express the antibody or antigen-binding fragment thereof encoded by the nucleic acid. The method may also include isolating the antibody or antigen-binding fragment thereof from the cell (or culture of cells) or from the medium in which the one or more cells are cultured.

[0096] In one aspect, the present disclosure provides a composition comprising any of the mature factor D-specific antibodies or antigen-binding fragments disclosed herein.

[0097] In one aspect, the present disclosure provides a substrate for use in an immunoassay comprising at least one or more of any of the mature factor D-specific antibodies or antigen-binding fragments disclosed herein.

[0098] In one aspect, the present disclosure provides a kit for detecting the presence or amount of mature factor D in a test sample, such as a biological sample, comprising: (a) at least one container; and (b) at least one or more of any of the mature factor D-specific antibodies or antigen-binding fragments disclosed herein.

[0099] B. Anti-human profactor D specific monoclonal antibody As described in Examples 8 and 9 herein, the inventors used the human propeptide "APPRGR" (SEQ ID NO:4), corresponding to residues 20-25 of human full-length factor D, as an antigen to generate anti-pro-factor D-specific antibodies suitable for use in the detection assays and methods described herein.

[0100] As described in Example 9, the variable heavy and light chain fragments of several representative anti-pro-factor D specific monoclonal antibodies were cloned and sequenced.

[0101] FIG. 16A is an amino acid sequence alignment of the variable heavy chain regions of six anti-pro-Factor D clones identified as having high binding affinity for the human Factor D propeptide "APPRGR" (SEQ ID NO:4).

[0102] FIG. 16B is an amino acid sequence alignment of the variable light chain regions of six anti-pro-Factor D clones identified as having high binding affinity for the human Factor D propeptide "APPRGR" (SEQ ID NO:4).

[0103] The heavy and light chain variable regions and CDRs therein of six pro-factor D-specific monoclonal antibodies are provided in Tables 3 and 4 below.

[0104] Table 3: Anti-human profactor D specific antibody sequences: Mouse parent TIFF2025076475000038.tif51139

[0105] Table 4. Anti-human profactor D specific antibodies: CDRs TIFF2025076475000039.tif51152

[0106] Thus, in one aspect, the present invention provides an isolated antibody or fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human Factor D designated as "APPRGR" (SEQ ID NO:4), wherein the antibody or fragment specifically binds to human pro-Factor D (SEQ ID NO:2) and not mature Factor D (SEQ ID NO:3). In one embodiment, the pro-Factor D-specific antibody is a monoclonal antibody. In one embodiment, the pro-Factor D-specific antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. In one embodiment, the pro-Factor D-specific antibody fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. In one embodiment, the pro-Factor D-specific antibody is a single-chain molecule. In one embodiment, the pro-Factor D-specific antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. In one embodiment, the pro-Factor D-specific antibody or antigen-binding fragment thereof has a K D In one embodiment, the pro-factor D-specific antibody or antigen-binding fragment thereof is labeled with a detectable moiety, e.g., a detectable moiety suitable for use in an immunoassay as further described herein. In one embodiment, the pro-factor D-specific antibody or fragment thereof is immobilized on a substrate, e.g., a substrate suitable for use in an immunoassay as further described herein.

[0107] In one embodiment, a pro-factor D-specific antibody or fragment thereof (i.e., an antibody or fragment thereof that specifically binds to human pro-factor D) comprises a binding domain comprising HC-CDR1, HC-CDR-2, and HC-CDR-3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136-141, and LC-CDR-1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142-147, wherein the CDRs are numbered according to the Kabat numbering system.

[0108] In one embodiment, a pro-factor D-specific antibody or fragment thereof that specifically binds to human pro-factor D comprises a binding domain comprising HC-CDR1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136-139, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142-145, wherein the CDRs are numbered according to the Kabat numbering system. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XYWMS (SEQ ID NO:201), wherein X at position 1 is N, S, or T; (b) amino acid sequence (c) HC-CDR2 comprising the amino acid sequence AWFAX (SEQ ID NO: 203) (wherein X at position 5 is S, Y, or N); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence WASTRES (SEQ ID NO: 178); and (f) an LC-CDR3 comprising the amino acid sequence LQYYXYPYT (SEQ ID NO: 205) (in which X at position 5 is T or S). In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 149 or SEQ ID NO: 155, (b) HC-CDR2 comprising SEQ ID NO: 151 or SEQ ID NO: 156; (c) HC-CDR3 comprising SEQ ID NO: 153; (d) LC-CDR1 comprising SEQ ID NO: 176, (e) LC-CDR2 comprising SEQ ID NO: 178, and (f) LC-CDR3 comprising SEQ ID NO: 180. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 136. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 137. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 142. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 143.In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 136 and a VL comprising SEQ ID NO: 142. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 137 and a VL comprising SEQ ID NO: 143.

[0109] In one embodiment, a pro-factor D-specific antibody or fragment thereof that specifically binds to human pro-factor D comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 158, (b) HC-CDR2 comprising SEQ ID NO: 159 or SEQ ID NO: 163; (c) HC-CDR3 comprising SEQ ID NO: 161 or SEQ ID NO: 165; (d) LC-CDR1 comprising SEQ ID NO: 184 or SEQ ID NO: 189, (e) LC-CDR2 comprising SEQ ID NO: 178, and (f) LC-CDR3 comprising SEQ ID NO: 187. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 138. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:139. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:144. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:145. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO:138, and a VL comprising SEQ ID NO:144. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO:139, and a VL comprising SEQ ID NO:145.

[0110] In one embodiment, a pro-factor D-specific antibody or fragment thereof that specifically binds to human pro-factor D comprises a binding domain comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NO: 140 and SEQ ID NO: 141, and LC-CDR1, LC-CDR2 and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO: 146 and SEQ ID NO: 147. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 167, (b) HC-CDR2 comprising SEQ ID NO: 169 or SEQ ID NO: 173, (c) HC-CDR3 comprising SEQ ID NO: 171 or SEQ ID NO: 174, (d) LC-CDR1 comprising SEQ ID NO: 194, (e) LC-CDR2 comprising SEQ ID NO: 196 or SEQ ID NO: 199, and (f) LC-CDR3 comprising SEQ ID NO: 198 or SEQ ID NO: 200. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO: 140. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:141. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:146. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:147.In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 140 and a VL comprising SEQ ID NO: 146. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a VH comprising SEQ ID NO: 141 and a VL comprising SEQ ID NO: 147.

[0111] In certain embodiments, a pro-factor D-specific antibody or fragment thereof has a heavy chain variable domain that is substantially identical (e.g., at least about 70%, at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least 99% identical) to any of the heavy chain variable domain sequences shown in Table 3. In certain embodiments, a pro-factor D-specific antibody or fragment thereof has a light chain variable domain that is substantially identical (e.g., at least about 70%, at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least 99% identical) to any of the light chain variable domain sequences shown in Table 3.

[0112] In another aspect, the disclosure provides a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy chain variable region of a pro-factor D-specific antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NOs: 136-141, and the CDRs are numbered according to the Kabat numbering system. In another aspect, the disclosure provides a nucleic acid encoding the complementarity determining regions (CDRs) of the light chain variable region of a pro-factor D-specific antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NOs: 142-147, and the CDRs are numbered according to the Kabat numbering system.

[0113] In another aspect, the disclosure provides a cloning or expression vector comprising a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 136-141, the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 142-147, and the CDRs are numbered according to the Kabat numbering system.

[0114] In another aspect, the disclosure provides a cell containing a cloning or expression vector comprising a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable regions of a pro-factor D-specific antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 136-141, the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 142-147, and the CDRs are numbered according to the Kabat numbering system.

[0115] In another aspect, the present disclosure provides a method for producing a human pro-factor D-specific antibody, comprising culturing cells containing an expression vector containing a nucleic acid encoding one or both of the heavy and light chain polypeptides of any of the pro-factor D-specific antibodies or antigen-binding fragments disclosed herein. The cell or culture of cells is cultured under conditions and for a time sufficient to allow the cell (or culture of cells) to express the antibody or antigen-binding fragment thereof encoded by the nucleic acid. The method may also include isolating the antibody or antigen-binding fragment thereof from the cell (or culture of cells) or from the medium in which the one or more cells are cultured.

[0116] In one aspect, the present disclosure provides a composition comprising any of the pro-factor D-specific antibodies or antigen-binding fragments disclosed herein.

[0117] In one aspect, the present disclosure provides a substrate for use in an immunoassay comprising at least one or more of any of the pro-factor D-specific antibodies or antigen-binding fragments disclosed herein.

[0118] In one aspect, the present disclosure provides a kit for detecting the presence or amount of pro-factor D in a test sample, such as a biological sample, comprising: (a) at least one container; and (b) at least one or more of any of the pro-factor D-specific antibodies or antigen-binding fragments disclosed herein.

[0119] C. Anti-human factor D monoclonal antibody that binds to both the pro- and mature forms of factor D As described in Examples 4-5 herein, the present invention also provides anti-factor D antibodies that bind to epitopes present in both human pro-factor D and human mature factor D. As described in Example 4, the inventors used human mature factor D (SEQ ID NO:3) as an antigen to generate anti-factor D antibodies that were screened and selected for their ability to detect both the pro- and mature forms of factor D and that are suitable for use in combination with the mature and pro-factor D-specific antibodies disclosed herein in the detection assays and methods described herein.

[0120] As described in Example 5, the variable heavy and light chain fragments of several representative anti-factor D monoclonal antibodies that bind to both pro- and mature factor D were cloned and sequenced.

[0121] FIG. 10A is an amino acid sequence alignment of the variable heavy chain regions of five anti-factor D clones identified as having high binding affinity for both mature and pro-factor D.

[0122] FIG. 10B is an amino acid sequence alignment of the variable light chain regions of five anti-factor D clones identified as having high binding affinity for both mature and pro-factor D.

[0123] The heavy and light chain variable regions and CDRs therein of five anti-factor D monoclonal antibodies that bind to both mature factor D and pro-factor D are provided in Tables 5 and 6 below.

[0124] Table 5: Anti-human factor D antibody sequences: mouse parent TIFF2025076475000042.tif55148

[0125] Table 6: Anti-human factor D antibodies: CDRs TIFF2025076475000043.tif56128

[0126] Thus, in one aspect, the present invention provides an isolated antibody or fragment thereof that specifically binds to an epitope present in both human profactor D (SEQ ID NO:2) and human mature factor D (SEQ ID NO:3), wherein the antibody comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs:85-88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs:89-93, wherein the CDRs are numbered according to the Kabat numbering system.

[0127] In one embodiment, the anti-factor D antibody that binds to an epitope present in both pro-factor D and mature factor D is a monoclonal antibody. In one embodiment, the anti-factor D antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. In one embodiment, the anti-factor D antibody fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. In one embodiment, the anti-factor D antibody is a single-chain molecule. In one embodiment, the anti-factor D antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. In one embodiment, the anti-factor D antibody or antigen-binding fragment thereof has a K of less than 10 nM. DIn one embodiment, the anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope present in both pro-factor D and mature factor D is labeled with a detectable moiety, e.g., a detectable moiety suitable for use in an immunoassay as described further herein. In one embodiment, the anti-factor D antibody or fragment thereof that binds to an epitope present in both pro-factor D and mature factor D is immobilized on a substrate, e.g., a substrate suitable for use in an immunoassay as described further herein.

[0128] In one embodiment, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D comprises a binding domain comprising HC-CDR1, HC-CDR2 and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2 and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, wherein the CDRs are numbered according to the Kabat numbering system.

[0129] In one embodiment, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO:95; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO:97; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO:99; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO:111; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO:113; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO:115.

[0130] In one embodiment, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO:101; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO:103 or 107; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO:105 or 108; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO:60 or 123; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO:119, 124 or 126; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO:121 or 125.

[0131] In one embodiment, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:85. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:89. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:90. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VH comprising SEQ ID NO:85 and a VL comprising SEQ ID NO:89. In one embodiment, the anti-factor D antibody or fragment thereof comprises a VH comprising SEQ ID NO:85, and a VL comprising SEQ ID NO:90.

[0132] In one embodiment, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:86. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:87. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VH domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:88.

[0133] In one embodiment, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:91. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:92. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VL domain having at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, or at least 99% identity) to the amino acid sequence of SEQ ID NO:93. In one embodiment, an anti-factor D antibody or fragment thereof comprises a VH comprising SEQ ID NO:86, and a VL comprising SEQ ID NO:91. In one embodiment, the anti-factor D antibody or fragment thereof comprises a VH comprising SEQ ID NO: 87 and a VL comprising SEQ ID NO: 92. In one embodiment, the anti-factor D antibody or fragment thereof comprises a VH comprising SEQ ID NO: 88 and a VL comprising SEQ ID NO: 93.

[0134] In certain embodiments, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D has a heavy chain variable domain that is substantially identical (e.g., at least about 70%, at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least 99% identical) to any of the heavy chain variable domain sequences shown in Table 5.

[0135] In certain embodiments, an anti-factor D antibody or fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D has a light chain variable domain that is substantially identical (e.g., at least about 70%, at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least 99% identical) to any of the light chain variable domain sequences shown in Table 5.

[0136] In another aspect, the disclosure provides a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy chain variable region of an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human maturation factor D and human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NOs:85-88, and the CDRs are numbered according to the Kabat numbering system. In another aspect, the disclosure provides a nucleic acid encoding the complementarity determining regions (CDRs) of the light chain variable region of an antibody or antigen-binding fragment thereof that binds to an epitope common to both human maturation factor D and human pro-factor D, wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NOs:89-93, and the CDRs are numbered according to the Kabat numbering system.

[0137] In another aspect, the present disclosure provides a cloning or expression vector comprising a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable regions of an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs:85-88, the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs:89-93, and the CDRs are numbered according to the Kabat numbering system.

[0138] In another aspect, the present disclosure provides a cell containing a cloning or expression vector comprising a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable regions of an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs:85-88, the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs:89-93, and the CDRs are numbered according to the Kabat numbering system.

[0139] In another aspect, the present disclosure provides a method for producing an anti-factor D antibody that binds to an epitope common to both human mature factor D and human pro-factor D, comprising culturing cells containing an expression vector containing nucleic acid encoding one or both of the heavy and light chain polypeptides of any of the antibodies or antigen-binding fragments disclosed herein. The cell or culture of cells is cultured under conditions and for a time sufficient to allow the cell (or culture of cells) to express the anti-factor D antibody or antigen-binding fragment thereof encoded by the nucleic acid. The method may also include isolating the antibody or antigen-binding fragment thereof from the cell (or culture of cells) or from the medium in which the one or more cells are cultured.

[0140] In one aspect, the present disclosure provides a composition comprising any of the anti-factor D antibodies or antigen-binding fragments thereof that bind to an epitope common to both human mature factor D and human pro-factor D as disclosed herein.

[0141] In one aspect, the present disclosure provides a substrate for use in an immunoassay comprising at least one or more of any of the anti-factor D antibodies or antigen-binding fragments thereof that bind to an epitope common to both human mature factor D and human pro-factor D disclosed herein.

[0142] In one aspect, the present disclosure provides a kit for detecting the presence of factor D in a test sample, such as a biological sample, comprising: (a) at least one container; and (b) at least one or more of any of the anti-factor D antibodies or antigen-binding fragments thereof that bind to an epitope common to both human mature factor D and human pro-factor D disclosed herein.

[0143] Single chain anti-factor D antibody In one embodiment of the present invention, the anti-factor D antibody (i.e., either a mature factor D-specific antibody, a pro-factor D-specific antibody, or an anti-factor D antibody that binds to both the mature and pro-forms of factor D) is a single-chain antibody, defined as a genetically engineered molecule containing the variable regions of the light chain and the variable regions of the heavy chain linked by a suitable polypeptide linker as a genetically fused single-chain molecule. Such single-chain antibodies are also referred to as "single-chain Fv" or "scFv" antibody fragments. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the scFv to form the desired structure for antigen binding. scFv antibodies that bind to factor D can be oriented so that the variable light chain region is either amino-terminal or carboxy-terminal to the variable heavy chain region.

[0144] Humanized anti-factor D antibody The anti-factor D antibodies disclosed herein (i.e., either mature factor D-specific antibodies, pro-factor D-specific antibodies, or anti-factor D antibodies that bind both mature and pro-forms of factor D) can be modified without altering their ability to be used for the purposes described herein. As a first issue, it is noted that the antibodies described herein are derived from immunized mice. Thus, the antibodies have framework regions (regions outside the complementarity-determining regions, or "CDRs") that contain amino acid residues typically found in the framework regions of mouse antibodies and may be immunogenic when administered to human patients. To reduce the immunogenicity of mouse antibodies for use in humans, it is common in the art to engineer the framework regions by replacing residues found at specific positions in mouse antibodies with residues more typically found at the same positions in human antibodies. Antibodies engineered in this way are referred to as "humanized antibodies" and are typically preferred for in vivo use because they have a lower risk of inducing side effects and typically remain in circulation longer. Methods for humanizing antibodies are known in the art and are described in detail, for example, in U.S. Patent Nos. 6,180,377; 6,407,213; 5,693,762; 5,585,089; and 5,530,101.

[0145] Furthermore, because the CDRs of the variable regions determine antibody specificity, the CDRs of the anti-factor D antibodies shown in Tables 2, 4, 6-10, and 12-17 can be grafted or engineered into an antibody of choice to confer specificity for binding to factor D. For example, CDRs from mature factor D-specific clones 6G6, 14A11, 27B3, 58F5, 49G3, and 10G1, as shown in Table 2, and / or CDRs from pro-factor D-specific clones 18F5, 1F9, 2A4, 20A1, 13A10, and 21H1, as shown in Table 4, can be grafted onto a human antibody framework of known three-dimensional structure (see, e.g., WO98 / 45322; Jones et al., Nature 321:522 (1986); Verhoeyen et al., Science 239:1534 (1988); Riechmann et al., Nature 332:323 (1988) and Winter & Milstein, Nature 349:293 (1991)), to generate anti-mature factor D-specific or anti-pro-factor D-specific antibodies that exhibit reduced or no immunogenic response when administered to humans.

[0146] Methods for producing anti-factor D antibodies In another aspect, the invention provides a method for producing an antibody that specifically recognizes and binds human factor D, e.g., a mature factor D-specific antibody, a pro-factor D-specific antibody, or an anti-factor D antibody that binds both the mature and pro-forms of factor D, comprising culturing cells containing an expression vector containing a nucleic acid encoding one or both of the heavy and light chain polypeptides of any of the antibodies or antigen-binding fragments disclosed herein. The cell or culture of cells is cultured under conditions and for a time sufficient to allow the cell (or culture of cells) to express the antibody or antigen-binding fragment thereof encoded by the nucleic acid. The method can also include isolating the antibody or antigen-binding fragment thereof from the cell (or culture of cells) or from the medium in which the one or more cells are cultured.

[0147] In one aspect, the disclosure features a cell containing a cloning or expression vector including a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs:12-17 and the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs:18-23.

[0148] In another aspect, the disclosure features a cell containing a cloning or expression vector including a nucleic acid encoding the complementarity determining regions (CDRs) of the heavy and / or light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, wherein the heavy chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 136-141 and the light chain variable region comprises the amino acid sequence set forth as any of SEQ ID NOs: 142-147.

[0149] In some embodiments, the invention provides nucleic acid molecules encoding anti-factor D antibodies or fragments thereof of the invention, e.g., antibodies or antigen-binding fragments thereof that specifically bind to human maturation factor D (e.g., as shown in Table 1), antibodies or antigen-binding fragments thereof that specifically bind to human pro-factor D (e.g., as shown in Table 3), or antibodies or antigen-binding fragments thereof that bind to an epitope common to both human maturation factor D and human pro-factor D (e.g., as shown in Table 5). In some embodiments, the invention provides nucleic acid molecules encoding anti-factor D antibodies, e.g., nucleic acid molecules comprising a nucleic acid sequence that encodes an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D (e.g., SEQ ID NOs:73-84), or that encodes an antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D (e.g., SEQ ID NOs:206-217), or that encodes an antibody or antigen-binding fragment thereof that binds to an epitope common to both human maturation factor D and human pro-factor D (e.g., SEQ ID NOs:127-135).

[0150] In some embodiments, the present invention provides cells containing nucleic acid molecules encoding factor D-specific monoclonal antibodies of the present invention (including mature factor D-specific antibodies, pro-factor D-specific antibodies, and anti-factor D antibodies that bind to both the mature and pro forms of factor D).

[0151] In some embodiments, the present invention provides expression cassettes comprising nucleic acid molecules encoding factor D-specific monoclonal antibodies of the present invention (including mature factor D-specific antibodies, pro-factor D-specific antibodies, and anti-factor D antibodies that bind to both the mature and pro forms of factor D).

[0152] In some embodiments, the present invention provides methods for producing factor D-specific monoclonal antibodies, comprising culturing cells containing nucleic acid molecules encoding factor D-specific antibodies of the present invention (including mature factor D-specific antibodies, pro-factor D-specific antibodies, and anti-factor D antibodies that bind to both the mature and pro forms of factor D).

[0153] In many embodiments, nucleic acids encoding the subject monoclonal antibodies are introduced directly into host cells, and the cells are incubated under conditions sufficient to induce expression of the encoded antibody.

[0154] In one embodiment, a method for producing a factor D-specific monoclonal antibody (including a mature factor D-specific antibody, a pro-factor D-specific antibody, or an anti-factor D antibody that binds to both the mature and pro forms of factor D) comprises culturing cells containing a nucleic acid molecule encoding a factor D-specific antibody of the present invention.

[0155] According to certain related aspects, there are provided recombinant host cells comprising one or more constructs as described herein; nucleic acids encoding any anti-factor D antibody, CDR, VH or VL domain, or antigen-binding fragment thereof; and methods for producing the encoded product, comprising expression from the encoding nucleic acid therefor. Expression can be conveniently achieved by culturing recombinant host cells containing the nucleic acid under appropriate conditions. Following production by expression, the antibody or antigen-binding fragment thereof can be isolated and / or purified using any suitable technique and then used as desired.

[0156] For example, any cell suitable for expressing the expression cassette may be used as a host cell, for example, yeast, insect, plant, etc. cells. In many embodiments, mammalian host cell lines that do not normally produce antibodies are used, examples of which include monkey kidney cells (COS cells), SV40-transformed monkey kidney CVI cells (COS-7, ATCC CRL 165 1); human embryonic kidney cells (HEK-293, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells (CHO, Urlaub and Chasin, Proc. Natl. Acad. Sci. (USA) 77:4216, (1980)); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CVI ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat hepatocytes (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human hepatocytes (hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL 51); TRI cells (Mather et al., Annals NY Acad. Sci 383:44-68 (1982)); NIH / 3T3 cells (ATCC CRL-1658); and mouse L cells (ATCC CCL-1). Additional cell lines will be apparent to those skilled in the art. A wide variety of cell lines are available from the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209.

[0157] Methods for introducing nucleic acid into cells are well known in the art. Suitable methods include electroporation, particle gun technology, calcium phosphate precipitation, direct microinjection, etc. The choice of method generally depends on the type of cell to be transformed and the situation in which transformation is carried out (i.e., in vitro, ex vivo, or in vivo). A general overview of these methods can be found in Ausubel, et al., Short Protocols in Molecular Biology, 3rd ed., Wiley & Sons, 1995. In some embodiments, lipofectamine and calcium-mediated gene transfer techniques are used.

[0158] After the subject nucleic acid has been introduced into the cells, the cells are typically incubated, usually at 37° C., for a suitable period of time, sometimes under selection, to allow for expression of the antibody. In most embodiments, the antibody is typically secreted into the supernatant of the medium in which the cells are growing.

[0159] In mammalian host cells, numerous virus-based expression systems can be used to express the subject antibody. When adenovirus is used as an expression vector, the antibody coding sequence of interest can be ligated to an adenovirus transcription / translation control complex, such as the late promoter and tripartite leader sequence. This chimeric gene can then be inserted into the adenovirus genome by in vitro or in vivo recombination. Insertion into a non-essential region of the viral genome (e.g., region E1 or E3) results in a recombinant virus that is viable in infected hosts and can express antibody molecules (see, e.g., Logan & Shenk, Proc. Natl. Acad. Sci. USA 81:355-359 (1984)). The efficiency of expression can be enhanced by incorporating appropriate transcription enhancer elements, transcription terminators, etc. (see, e.g., Bittner et al., Methods in Enzymol. 153:51-544 (1987)).

[0160] For long-term, high-yield production of recombinant antibodies, stable expression may be used. For example, cell lines that stably express antibody molecules may be engineered. Rather than using expression vectors containing viral origins of replication, host cells can be transformed with an immunoglobulin expression cassette and a selectable marker. Following the introduction of the foreign DNA, the engineered cells may be grown for 1-2 days in an enriched medium and then switched to a selective medium. The selectable marker in the recombinant plasmid confers resistance to the selection and allows the cells to stably integrate the plasmid into their chromosomes and grow to form foci, which can then be cloned and expanded into cell lines. Such engineered cell lines may be particularly useful in screening and evaluation of compounds that interact directly or indirectly with the antibody molecule.

[0161] Once the antibody molecules of the present invention are produced, they may be purified by any method known in the art for the purification of immunoglobulin molecules, such as, for example, chromatography (e.g., ion exchange, affinity, particularly for a specific antigen followed by protein A, and sizing column chromatography), centrifugation, differential solubility, or any other standard technique for protein purification. In many embodiments, the antibody is secreted from the cells into the culture medium and collected from the culture medium. For example, a nucleic acid sequence encoding a signal peptide may be incorporated adjacent to the coding region of the antibody or fragment. Such a signal peptide can be incorporated adjacent to the 5' end of the amino acid sequence set forth herein for the subject antibodies to facilitate production of the subject antibodies.

[0162] Anti-factor D antibody labeled with a detectable moiety In another aspect, the present invention provides anti-factor D antibodies (including mature factor D-specific antibodies, pro-factor D-specific antibodies, and anti-factor D antibodies that bind to both the mature and pro-forms of factor D) labeled with a detectable moiety (i.e., a moiety that allows for detection and / or quantification). In various embodiments, the antibodies described herein are conjugated to a detectable label that can be detected directly or indirectly. In this regard, an antibody "conjugate" refers to an anti-factor D antibody covalently linked to a detectable label. In the present invention, monoclonal antibodies, antigen-binding fragments thereof, and antibody derivatives thereof, such as single-chain variable fragment antibodies or epitope-tagged antibodies, may all be covalently linked to a detectable label. In "direct detection," only one detectable antibody, i.e., a primary detectable antibody, is used. Thus, direct detection means that an antibody conjugated to a detectable label can itself be detected without the need for the addition of a second antibody (secondary antibody).

[0163] A "detectable label" is a molecule or substance that can generate a detectable (e.g., visually, electronically, or otherwise) signal that indicates the presence and / or concentration of the label in a sample. When conjugated to an antibody, a detectable label can be used to locate and / or quantify the target to which the specific antibody is directed. The presence and / or concentration of the target in a sample can thereby be detected by detecting the signal generated by the detectable label. Detectable labels can be detected directly or indirectly, and several different detectable labels conjugated to different specific antibodies can be used in combination to detect one or more targets.

[0164] Examples of detectable labels that can be directly detected include fluorescent dyes, radioactive substances, and metal particles.In contrast, indirect detection requires the application of one or more additional antibodies, i.e., secondary antibodies, after the application of a primary antibody.Therefore, the detection is carried out by detecting the binding of a secondary antibody or binding substance to the primary detectable antibody.Examples of primary detectable binding substances or antibodies that require the addition of a secondary binding substance or antibody include enzymatically detectable binding substances and hapten-detectable binding substances or antibodies.

[0165] Examples of detectable labels that can be conjugated to the antibodies of the present disclosure include fluorescent labels, enzyme labels, radioisotopes, chemiluminescent labels, electrochemiluminescent labels, bioluminescent labels, polymers, polymer particles, metal particles, haptens, and dyes.

[0166] Examples of fluorescent labels include 5-(and 6)-carboxyfluorescein, 5- or 6-carboxyfluorescein, 6-(fluorescein)-5-(and 6)-carboxamidohexanoic acid, fluorescein isothiocyanate, rhodamine, tetramethylrhodamine, and dyes such as Cy2, Cy3, and Cy5, optionally substituted coumarins (including AMCA), PerCP, phycobiliproteins (including R-phycoerythrin (RPE) and allophycoerythrin (APC)), Texas Red, Princeton Red, green fluorescent protein (GFP) and analogs thereof, and conjugates of R-phycoerythrin or allophycoerythrin, inorganic fluorescent labels, for example, particles based on semiconductor materials such as coated CdSe nanocrystals.

[0167] Examples of polymer particle labels include polystyrene, PMMA or silica microparticles or latex particles into which fluorescent dyes can be embedded, or polymer micelles or capsules containing dyes, enzymes or substrates.

[0168] Examples of metal particle labels include gold particles and coated gold particles that can be converted by silver staining. Examples of haptens include DNP, fluorescein isothiocyanate (FITC), biotin, and digoxigenin. Examples of enzyme labels include horseradish peroxidase (HRP), alkaline phosphatase (ALP or AP), β-galactosidase (GAL), glucose-6-phosphate dehydrogenase, β-N-acetylglucosaminidase, β-glucuronidase, invertase, xanthine oxidase, firefly luciferase, and glucose oxidase (GO). Examples of commonly used substrates for horseradish peroxidase include 3,3'-diaminobenzidine (DAB), nickel-enhanced diaminobenzidine, 3-amino-9-ethylcarbazole (AEC), benzidine dihydrochloride (BDHC), Hanker-Yates reagent (HYR), indophane blue (IB), tetramethylbenzidine (TMB), 4-chloro-1-naphthol (CN), alpha-naphtholpyronine (ALPHA), and alpha-naphtholpyronine (ALPHA). Examples of suitable antibacterial agents include 5-bromo-4-chloro-3-indolyl phosphate (BCIP), o-dianisidine (OD), 5-bromo-4-chloro-3-indolyl phosphate (BCIP), nitroblue tetrazolium (NBT), 2-(p-iodophenyl)-3-p-nitrophenyl-1-5-phenyltetrazolium chloride (INT), tetranitroblue tetrazolium (TNBT), and 5-bromo-4-chloro-3-indoxyl-beta-D-galactoside / ferro-ferricyanide (BCIG / FF).

[0169] Examples of commonly used substrates for alkaline phosphatase include naphthol-AS-B 1-phosphate / Fast Red TR (NABP / FR), naphthol-AS-MX-phosphate / Fast Red TR (NAMP / FR), naphthol-AS-B 1-phosphate / Fast Red TR (NABP / FR), naphthol-AS-MX-phosphate / Fast Red TR (NAMP / FR), naphthol-AS-B 1-phosphate / New Fuchsin (NABP / NF), bromochloroindolyl phosphate / nitroblue tetrazolium (BCIP / NBT), and 5-bromo-4-chloro-3-indolyl-b-d-galactopyranoside (BCIG).

[0170] Examples of luminescent labels include luminol, isoluminol, acridinium esters, 1,2-dioxetanes, and pyridopyridazines. Examples of electrochemiluminescent labels include ruthenium derivatives. Examples of radioactive labels include radioisotopes of iodide, cobalt, selenium, tritium, carbon, sulfur, and phosphorus.

[0171] The detectable label may be linked to the antibody described herein (i.e., a mature factor D-specific antibody, a pro-factor D-specific antibody, or an anti-factor D antibody that binds to both the mature and pro-forms of factor D) or to any other molecule that specifically binds to the biological marker of interest, such as an antibody, a nucleic acid probe, or a polymer. Furthermore, those skilled in the art will recognize that the detectable label can also be conjugated to a second, third, fourth, and / or fifth binding substance or antibody, etc. Furthermore, those skilled in the art will recognize that each additional binding substance or antibody used to characterize the biological marker of interest can serve as a signal amplification step. When the detectable substance is, for example, a dye, colloidal gold particles, or a luminescent reagent, the biological marker can be visually detected using, for example, optical microscopy, fluorescence microscopy, or electron microscopy. Visually detectable substances bound to biological markers can also be detected using a spectrophotometer. If the detectable substance is a radioisotope, detection can be accomplished visually by autoradiography or non-visually using a scintillation counter. See, e.g., Larsson, 1988, Immunocytochemistry: Theory and Practice, (CRC Press, Boca Raton, Fla.); Methods in Molecular Biology, vol. 80 1998, John D. Pound (ed.) (Humana Press, Totowa, NJ).

[0172] In another embodiment, the anti-factor D antibody is unlabeled (i.e., naked), and its presence can be detected using a labeled antibody that binds to the anti-factor D antibody (i.e., either a mature factor D-specific antibody, a pro-factor D-specific antibody, or an anti-factor D antibody that binds to both the mature and pro forms of factor D).

[0173] V. Compositions and Kits Comprising Anti-Factor D Antibodies composition In another aspect, the present disclosure provides a substrate, e.g., a solid support (e.g., an insoluble substrate, e.g., a non-aqueous matrix such as a glass plate or slide, a polysaccharide (e.g., agarose), polyacrylamide, polystyrene, plastic or metal, a polymer-coated bead, a tube, or a ceramic or metal chip), comprising immobilized (or otherwise deposited) a monoclonal anti-factor D antibody disclosed herein (e.g., a mature factor D-specific antibody, a pro-factor D-specific antibody, and an anti-factor D antibody that binds to both the mature and pro-forms of factor D). In some embodiments, the anti-factor D antibodies are immobilized (or deposited) at separate locations (e.g., in the wells of a multi-wall plate or in an array on a biochip). In some embodiments, a substrate comprising anti-factor D antibodies can be part of a kit for detecting factor D (e.g., mature factor D, pro-factor D, or total factor D (mature and pro-factor D)) in a biological sample obtained from a mammalian subject.

[0174] kit In another aspect, the present disclosure provides kits for use in performing one or more of the assays disclosed herein.

[0175] In one aspect, the present disclosure provides kits (i.e., packaged combinations of reagents in predetermined amounts) containing reagents and instructions for detecting the presence of factor D (e.g., mature factor D, pro-factor D, or total factor D (mature and pro-factor D)) in a test sample, such as a biological sample. Exemplary kits may contain at least one anti-factor D monoclonal antibody or antigen-binding fragment thereof as described herein (i.e., either a mature factor D-specific antibody, a pro-factor D-specific antibody, or an anti-factor D antibody that binds to both the mature and pro-forms of factor D). If the anti-factor D antibody is labeled with a detectable moiety, such as an enzyme, the kit will include substrates and cofactors required for the enzyme (e.g., a substrate precursor that provides a detectable chromophore or fluorophore). In addition, other additives, such as stabilizers, buffers (e.g., blocking buffers or cell lysis buffers), and the like, may be included. The relative amounts of the various reagents may vary widely to provide concentrations in solution of the reagents that substantially optimize the sensitivity of the assay. Notably, the reagents may be provided as dry powders, usually lyophilized, containing excipients that, upon dissolution, provide a reagent solution having the appropriate concentrations.

[0176] In addition, the kit may include instructional materials disclosing means of use of the antibodies of the invention (e.g., for detection of mature factor D or pro-factor D as a biomarker for the level or absence of alternative pathway complement (APC) activation). The kit may also include additional components to facilitate the particular application for which the kit is designed. For example, the kit may additionally contain means for detecting the label (e.g., an enzyme substrate for an enzymatic label, a filter set for detecting a fluorescent label, an appropriate secondary label such as sheep anti-mouse-HRP, etc.). The kit may additionally include buffers and other reagents routinely used in performing a particular immunoassay, as is well known in the art.

[0177] Certain embodiments provide kits for detecting the presence or amount of mature factor D in a sample, containing at least one mature factor D-specific antibody as described herein, e.g., an antibody or fragment comprising a CDR from mature factor D-specific clones 6G6, 14A11, 27B3, 58F5, 49G3, and 10G1 as shown in Table 2. In certain embodiments, the kit may include buffers, enzymes, labels, substrates, beads, or other surfaces to which the antibodies of the invention may be attached, and instructions for use.

[0178] Certain embodiments provide kits for detecting the presence or amount of pro-factor D in a sample, the kits containing at least one pro-factor D-specific antibody as described herein, e.g., an antibody or fragment comprising a CDR from pro-factor D-specific clones 18F5, 1F9, 2A4, 20A1, 13A10, and 21H1 as shown in Table 4. The subject anti-factor D antibodies and antigen-binding fragments thereof can be labeled with any suitable detectable moiety as described herein. In certain embodiments, the kits can include buffers, enzymes, labels, substrates, beads, or other surfaces to which the antibodies of the invention can be attached, and instructions for use.

[0179] The items in the kit may be individually wrapped or packaged in individual containers that are provided together within a larger container (e.g., a cardboard or Styrofoam box).

[0180] In accordance with the foregoing, in one aspect, the present disclosure provides a kit comprising at least one monoclonal antibody that specifically detects or quantifies human maturation factor D (SEQ ID NO:3) and / or pro-factor D (SEQ ID NO:2) in an immunoassay, wherein the at least one monoclonal antibody comprises: (i) a maturation factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope encompassing the amino terminus of human maturation factor D (the epitope comprises or consists of amino acids ILGGREA (SEQ ID NO:5) and the antibody does not bind to human pro-factor D (SEQ ID NO:2)); and / or (ii) a pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope on the activation ("pro") peptide of human factor D (the epitope comprises or consists of "APPRGR" (SEQ ID NO:4) and the antibody does not bind to maturation factor D (SEQ ID NO:3)). In one embodiment, a mature factor D-specific antibody or fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, wherein the CDRs are numbered according to the Kabat numbering system. In one embodiment, a pro-factor D-specific antibody or fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, wherein the CDRs are numbered according to the Kabat numbering system.

[0181] In some embodiments, the kit further comprises an anti-factor D antibody, or fragment thereof, that binds to an epitope common to both human mature factor D (SEQ ID NO:3) and human pro-factor D (SEQ ID NO:2). In some embodiments, the anti-factor D antibody or fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs:85-88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs:89-93, wherein the CDRs are numbered according to the Kabat numbering system.

[0182] In some embodiments, the kit further comprises at least one container.

[0183] In some embodiments, the kit is for performing an enzyme-linked immunosorbent assay (ELISA). In one embodiment, the mature factor D-specific antibody or fragment thereof is a coating antibody. In one embodiment, the mature factor D-specific antibody or fragment thereof is a detection antibody. In one embodiment, the pro-factor D-specific antibody or fragment thereof is a coating antibody. In one embodiment, the pro-factor D-specific antibody or fragment thereof is a detection antibody.

[0184] In various embodiments of the kits of the invention, the subject anti-factor D antibodies and antigen-binding fragments thereof (i.e., mature factor D-specific antibodies, pro-factor D-specific antibodies and / or anti-factor D antibodies) can be labeled with any suitable detectable moiety as described herein. In certain embodiments, the kits further include buffers, enzymes, labels, substrates, beads, or other surfaces to which the antibodies of the invention can be attached, and instructions for use.

[0185] VI. Methods for detecting factor D using anti-factor D antibodies As described herein, the inventors have generated anti-factor D antibodies suitable for use in immunoassays to detect the presence and / or quantity of factor D (e.g., mature factor D, pro-factor D, and total factor D (both mature and pro-forms of factor D)) in a test sample, such as a biological sample obtained from a mammalian subject.

[0186] In one aspect, the anti-factor D antibodies of the present invention (including mature factor D-specific antibodies, pro-factor D-specific antibodies, and anti-factor D antibodies that bind to both the mature and pro-forms of factor D) are used in in vitro immunoassays to analyze a test sample, e.g., a biological sample obtained from a test subject, for the presence or amount of pro-factor D, mature factor D, and / or total factor D. In such in vitro immunoassays, the anti-factor D antibodies or antigen-binding fragments thereof can be naked or labeled with a detectable moiety as described herein, and can be utilized in solution phase or bound to a substrate as described below. For in vitro assays, any type of antibody can be utilized, such as murine, chimeric, humanized, or human, since there is no need to consider the host immune response.

[0187] The antibodies of the present disclosure may be used in any known immunoassay method, such as competitive binding assays, direct and indirect sandwich assays, lateral flow assays (e.g., dipstick format), and immunoprecipitation assays (see, e.g., Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press. Inc., 1987)).

[0188] Sandwich assays involve the use of two antibodies, each capable of binding to a different immunogenic portion or epitope of the protein (e.g., factor D) to be detected. In a sandwich assay, the test sample analyte binds to a first antibody (e.g., an anti-factor D antibody, e.g., an antibody specific for mature factor D, an antibody specific for pro-factor D, and / or an antibody that binds both mature factor D and pro-factor D) immobilized on a solid support (e.g., a substrate), and then the second antibody binds to the analyte, thereby forming an insoluble tripartite complex. The second antibody may itself be labeled with a detectable moiety (direct sandwich assay) or may be measured using an anti-immunoglobulin antibody labeled with a detectable moiety (indirect sandwich assay).

[0189] For example, one preferred type of sandwich assay is an ELISA assay, in which the detectable moiety is an enzyme. Regardless of the detection system used, an ELISA assay generally involves immobilizing an antigen or antibody on a substrate (e.g., a solid support) and using an appropriate detection reagent. In an ELISA assay, a protein antigen-antibody reaction occurs on a substrate (e.g., a solid support), typically in a well on a microtiter plate. The antigen reacts with this first antibody (also called a coating or capture antibody) to form a stable complex, which can be visualized by adding a second antibody (called a detection antibody), which can be directly or indirectly linked to an enzyme. Addition of a substrate for the enzyme produces color, which can be measured photometrically.

[0190] In one embodiment, the anti-factor D antibodies of the present invention (including mature factor D-specific antibodies, pro-factor D-specific antibodies, and anti-factor D antibodies that bind to both the mature and pro forms of factor D) are used to detect the presence of the mature or pro form of factor D antigen in a biological sample using an enzyme-linked immunosorbent assay (ELISA) (see, e.g., Gold et al. J Clin Oncol. 24:252-58, 2006).

[0191] In direct competitive ELISA, a pure or semi-pure antigen preparation is bound to a substrate that is insoluble in the fluid or cell extract being tested, and a fixed amount of detectably labeled soluble antibody is added, allowing for detection and / or quantification of the binary complex formed between the substrate-bound antigen and the labeled antibody.

[0192] In contrast, "double-determinant" ELISA, also known as "two-site ELISA" or "sandwich assay," requires only small amounts of antigen, and the assay does not require extensive purification of the antigen. Therefore, double-determinant ELISA is preferable to direct competitive ELISA for detecting antigens in clinical samples. See, for example, the use of double-determinant ELISA for the quantification of c-myc oncoprotein in biopsy specimens (Field et al., Oncogene 4: 1463 (1989); Spandidos et al., AntiCancer Res. 9: 821 (1989)). In double-determinant ELISA, a fixed amount of unlabeled monoclonal antibody or antibody fragment ("capture antibody") is bound to a substrate (e.g., a solid support), the test sample is contacted with the capture antibody, and a fixed amount of detectably labeled soluble antibody (or antibody fragment) is added, allowing for detection and / or quantification of the ternary complex formed between the capture antibody, antigen, and labeled antibody.

[0193] In one embodiment, the capture antibody bound to the substrate (e.g., solid support) is an anti-factor D antibody or antigen-binding fragment thereof as disclosed herein that binds to an epitope common to both pro-factor D and mature factor D (i.e., in the C-terminal portion of factor D). In one embodiment, the capture antibody bound to the substrate (e.g., solid support) is a mature factor D-specific antibody or antigen-binding fragment thereof as disclosed herein. In one embodiment, the capture antibody bound to the substrate (e.g., solid support) is a pro-factor D-specific antibody or antigen-binding fragment thereof as disclosed herein.

[0194] Methods for performing double-determinant ELISA are well known to those skilled in the art. See, for example, Field et al., Oncogene 4: 1463 (1989); Spandidos et al., AntiCancer Res. 9: 821 (1989); and Moore et al., Methods in Molecular Biology Vol 10:273-281 (The Humana Press, Inc. 1992).

[0195] In a double-determinant ELISA, the soluble antibody or antibody fragment must bind to a factor D epitope that is distinct from the epitope recognized by the capture antibody. Double-determinant ELISA can be performed to determine whether factor D antigen (i.e., mature factor D or pro-factor D) is present in a test biological sample, such as a body fluid (e.g., blood, plasma, or serum) or a biopsy sample. Alternatively, the assay can be performed to quantify the amount of factor D antigen present in a clinical sample of body fluid. Quantitative assays can be performed by including dilutions of purified factor D antigen.

[0196] In vitro immunoassays can be performed in which at least one anti-factor D antibody or antigen-binding fragment thereof (e.g., a mature factor D-specific antibody, a pro-factor D-specific antibody, and / or an anti-factor D antibody that binds to both the mature and pro-forms of factor D) is bound to a substrate (e.g., a solid-phase support). For example, to link the monoclonal antibody to an insoluble substrate such as a polymer-coated bead, plate, tube, or ceramic or metal chip, the anti-factor D monoclonal antibody or fragment thereof can be attached to a polymer such as aminodextran. In one embodiment, the substrate is suitable for use in an ELISA method (e.g., a multiwell microtiter plate). Thus, determining the level of factor D (e.g., mature factor D, pro-factor D, or both mature factor D and pro-factor D) in a sample can be performed by commercially available methods such as ELISA bead assays, chemical or enzymatic protein measurements, etc.

[0197] Other suitable in vitro assays will be readily apparent to those skilled in the art. The specific concentration of the detectably labeled anti-anti-factor D antibody, the temperature and time of incubation, and other assay conditions may vary depending on various factors, including the concentration of factor D antigen in the sample, the nature of the sample, etc. The binding activity of the anti-factor D antibody sample can be determined according to well-known methods. Those skilled in the art will be able to determine the optimal and operable assay conditions for each determination by routine experimentation.

[0198] In another embodiment, the subject antibodies and antigen-binding fragments thereof can be used to detect the presence of factor D antigen in tissue sections prepared from tissue specimens (e.g., biopsy samples). Such in situ detection can be used to determine the presence of factor D antigen and to determine the distribution of factor D antigen in the examined tissue. In situ detection can be achieved by applying a detectably labeled anti-factor D antibody to the tissue section. General techniques for in situ detection are well known to those skilled in the art. See, for example, Ponder, "Cell Marking Techniques and Their Application," in Mammalian Development: A Practical Approach 113-38 Monk (ed.) (IRL Press 1987).

[0199] A. Assay for detecting maturation factor D In accordance with the foregoing, in one aspect, the present invention provides a method for determining the presence or amount of mature factor D in a test sample, such as a biological sample, comprising: (a) contacting the test sample with a mature factor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or absence of binding of the antibody, wherein the presence of binding indicates the presence or amount of mature factor D in the sample. In one embodiment, the mature factor D-specific antibody or fragment thereof binds to an epitope in the amino-terminal region of human mature factor D, wherein the epitope comprises or consists of amino acids ILGGREA (SEQ ID NO:5), and the antibody does not bind to pro-factor D.

[0200] In one embodiment, the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23. In one embodiment, the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO:65), in which X at position 1 is T, I, or S, and X at position 3 is G or I; (b) amino acid sequence HC-CDR2 containing TIFF2025076475000044.tif4128 (in the sequence, X at position 11 is H or N, and X at position 16 is S or R); (c) amino acid sequence HC-CDR3 containing TIFF2025076475000045.tif4128 (in the sequence, X at position 6 is R, G, or N, X at position 7 is S or Y, X at position 8 is F, I, or V, and X at position 10 is D or H); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69) (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54). In one embodiment, the anti-human maturation factor D specific antibody or antigen-binding fragment thereof comprises a binding domain comprising the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:25, (b) HC-CDR2 comprising SEQ ID NO:27; (c) HC-CDR3 comprising SEQ ID NO:29; (d) LC-CDR1 comprising SEQ ID NO:50, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54.

[0201] In some embodiments, the anti-human maturation factor D specific antibody or fragment thereof is a monoclonal antibody comprising CDRs from maturation factor D specific clones 6G6, 14A11, 27B3, 58F5, 49G3 and 10G1 as shown in Table 2.

[0202] In one embodiment, the method further comprises comparing the amount of maturation factor D detected according to step (b) to a reference standard or control sample to determine the level of maturation factor D in the test sample.

[0203] In one embodiment, the control sample is an individual or pooled sample from a subject suffering from an alternative pathway disease or disorder (e.g., paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, or other alternative pathway disease or disorder). In one embodiment, the control sample is an individual or pooled sample from a normal healthy volunteer. In one embodiment, the control sample is a baseline sample from a subject before treatment with a complement inhibitor (e.g., a MASP-3 inhibitor or other complement inhibitor). In one embodiment, the reference standard is at least one ratio of pro-factor D to mature factor D, or mature factor D to total factor D, obtained from a test sample or a control sample (e.g., an individual or pooled sample from a normal healthy volunteer, or a baseline sample from a subject before treatment with a complement inhibitor, or an individual or pooled sample from a subject suffering from an alternative pathway disease or disorder). In one embodiment, the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof is immobilized on a substrate. In one embodiment, the immunoassay is an ELISA assay.

[0204] In one embodiment, the anti-human maturation factor D-specific antibody is labeled with a detectable moiety, and step (b) comprises detecting the presence of the detectable moiety. In one embodiment, the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof is naked (i.e., unlabeled), and the presence or amount of antibody or fragment thereof bound to maturation factor D is detected using a labeled antibody that binds to the anti-maturation factor D antibody. In one embodiment, the anti-human maturation factor D-specific antibody or antigen-binding fragment thereof is immobilized on a substrate (i.e., captured / coated), and bound maturation factor D is detected with a second antibody that binds to a different epitope of factor D (e.g., an anti-factor D antibody that binds to an epitope common to maturation factor D and pro-factor D as described herein).

[0205] In one embodiment, the test sample is a biological sample obtained from a mammalian subject. In various embodiments, the biological sample is selected from the group consisting of whole blood, serum, plasma, sputum, amniotic fluid, cerebrospinal fluid, cell lysate, ascites, urine, saliva, and tissue. In one embodiment, the biological sample is selected from the group consisting of blood, serum, plasma, urine, and cerebrospinal fluid.

[0206] In one embodiment, the mammalian subject (e.g., a human) has or is at risk of developing an alternative pathway disease or disorder. In one embodiment, the mammalian subject has or develops a renal disease in which reduced renal function impairs complement factor D clearance.

[0207] In one aspect, the mammalian subject (e.g., a human) is being treated with a complement inhibitor, such as an alternative pathway complement inhibitor, e.g., a MASP-3 inhibitor (e.g., a MASP-3 inhibitory antibody), as further described herein.

[0208] As described herein, methods for detecting mature factor D according to various aspects of the present disclosure can be used to define the pharmacodynamic endpoint or therapeutic threshold of complement inhibitors, e.g., alternative pathway complement inhibitors, e.g., MASP-3 inhibitors (e.g., MASP-3 inhibitory antibodies).

[0209] Although the details of the immunoassay may vary depending on the particular format used, a method for detecting mature factor D in a test sample includes contacting the test sample with an antibody that specifically binds to mature factor D. The antibody is allowed to bind to mature factor D in the sample under immunologically reactive conditions, and the presence of the bound antibody is detected directly or indirectly. The mature factor D-specific antibody can be used, for example, as a capture antibody in an ELISA, or as a second antibody to bind to mature factor D captured by the capture antibody. As is known in the art, the presence of the second antibody is typically then detected. In some embodiments, the immunoassay is performed on a solid support. In some embodiments, the immunoassay is an ELISA assay.

[0210] B. Profactor D Assay In accordance with the foregoing, in another aspect, the present invention provides a method for detecting the presence or amount of pro-factor D in a test sample, comprising: (a) contacting the test sample with a pro-factor D-specific antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or absence of binding of the antibody, wherein the presence of binding indicates the presence of pro-factor D in the sample. In one embodiment, the pro-factor D-specific antibody or fragment thereof specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) of human factor D, wherein the antibody or fragment thereof specifically binds to human pro-factor D (SEQ ID NO:2) and does not bind to human mature factor D (SEQ ID NO:3).

[0211] In one embodiment, a pro-factor D-specific antibody or fragment thereof that specifically binds to human pro-factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136-141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142-147, wherein the CDRs are numbered according to the Kabat numbering system. In one embodiment, a pro-factor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136-139, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142-145, wherein the CDRs are numbered according to the Kabat numbering system. In one embodiment, the pro-factor D-specific antibody or fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NO: 140 and SEQ ID NO: 141, and LC-CDR-1, LC-CDR2 and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO: 146 and SEQ ID NO: 147, wherein the CDRs are numbered according to the Kabat numbering system.

[0212] In one embodiment, the pro-factor D-specific antibody or antigen-binding fragment thereof comprises a binding domain comprising the following six CDRs: (a) CDR-H1 comprising SEQ ID NO:167, (b) CDR-H2 comprising SEQ ID NO:169 or SEQ ID NO:173; (c) CDR-H3 comprising SEQ ID NO:171 or SEQ ID NO:174; (d) CDR-L1 comprising SEQ ID NO:194, (e) CDR-L2 comprising SEQ ID NO:196 or SEQ ID NO:199, and (f) CDR-L3 comprising SEQ ID NO:198 or SEQ ID NO:200.

[0213] In some embodiments, the pro-factor D-specific antibody or fragment thereof is a monoclonal antibody comprising CDRs from pro-factor D-specific clones 18F5, 1F9, 2A4, 20A1, 13A10, and 21H1 as shown in Table 4.

[0214] In one embodiment, the method further comprises comparing the amount of pro-factor D detected according to step (b) with a reference standard or control sample to determine the level of pro-factor D in the test sample.

[0215] In one embodiment, the control sample is an individual or pooled sample from a subject suffering from an alternative pathway disease or disorder (e.g., paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, or other alternative pathway disease or disorder). In one embodiment, the control sample is an individual or pooled sample from a normal healthy volunteer. In one embodiment, the control sample is a baseline sample from a subject before treatment with a complement inhibitor (e.g., a MASP-3 inhibitor or other complement inhibitor). In one embodiment, the reference standard is at least one ratio of pro-factor D to mature factor D, or pro-factor D to total factor D, obtained from a test sample or a control sample (e.g., an individual or pooled sample from a normal healthy volunteer, or a baseline sample from a subject before treatment with a complement inhibitor, or an individual or pooled sample from a subject suffering from an alternative pathway disease or disorder).

[0216] In one embodiment, the anti-human pro-factor D-specific antibody or antigen-binding fragment thereof is immobilized on a substrate. In one embodiment, the immunoassay is an ELISA assay.

[0217] In one embodiment, the anti-human pro-factor D-specific antibody is labeled with a detectable moiety, and step (b) comprises detecting the presence of said detectable moiety. In one embodiment, the anti-human pro-factor D-specific antibody or antigen-binding fragment thereof is naked (i.e., unlabeled), and the presence or amount of the antibody or fragment thereof bound to pro-factor D is detected using a labeled antibody that binds to the anti-human pro-factor D antibody. In one embodiment, the anti-human pro-factor D-specific antibody or antigen-binding fragment thereof is immobilized on a substrate (i.e., captured / coated), and bound pro-factor D is detected with a second antibody that binds to a different epitope of factor D (e.g., an anti-factor D antibody that binds to an epitope common to mature factor D and pro-factor D as described herein).

[0218] In one embodiment, the test sample is a biological sample obtained from a mammalian subject. In various embodiments, the biological sample is selected from the group consisting of whole blood, serum, plasma, sputum, amniotic fluid, cerebrospinal fluid, cell lysate, ascites, urine, saliva, and tissue. In one embodiment, the biological sample is selected from the group consisting of blood, serum, plasma, urine, and cerebrospinal fluid.

[0219] In one embodiment, the mammalian subject (e.g., a human) has or is at risk of developing an alternative pathway disease or disorder. In one embodiment, the mammalian subject has or develops a renal disease in which reduced renal function impairs complement factor D clearance.

[0220] In one aspect, the mammalian subject (e.g., a human) is being treated with a complement inhibitor, such as an alternative pathway complement inhibitor, e.g., a MASP-3 inhibitor (e.g., a MASP-3 inhibitory antibody), as further described herein.

[0221] As described herein, methods for detecting pro-factor D according to various embodiments of the present disclosure can be used to define the pharmacodynamic endpoint or therapeutic threshold of a complement inhibitor, e.g., an alternative pathway complement inhibitor, e.g., a MASP-3 inhibitor (e.g., a MASP-3 inhibitory antibody). In one embodiment, a mammalian subject (e.g., a human) is being treated with a MASP-3 inhibitor, e.g., a MASP-3 inhibitory antibody, as further described herein.

[0222] Although the details of the immunoassay may vary depending on the particular format used, methods for detecting pro-factor D in a test sample include contacting the test sample with an antibody that specifically binds to pro-factor D. The antibody is allowed to bind to pro-factor D in the sample under immunologically reactive conditions, and the presence of the bound antibody is detected directly or indirectly. The pro-factor D-specific antibody can be used, for example, as a capture antibody in an ELISA, or as a second antibody to bind to pro-factor D captured by the capture antibody. As is known in the art, the presence of the second antibody is typically then detected. In some embodiments, the immunoassay is performed on a solid support. In some embodiments, the immunoassay is an ELISA assay.

[0223] VII. METHODS OF DIAGNOSIS, MONITORING AND TREATMENT OF SUBJECTS SUFFERING FROM OR AT RISK OF DEVELOPING ALTERNATIVE PATHWAY DISEASES OR DISORDERS The anti-factor D antibodies, methods, reagents, and kits of the present invention can be used in numerous applications. For example, in certain embodiments, the assays of the present invention can be used to assess the level of mature factor D and / or pro-factor D in a subject and / or to assess the extent to which a complement pathway inhibitor, e.g., an alternative pathway complement inhibitor, e.g., a MASP-3 inhibitor (e.g., a MASP-3 inhibitory antibody), affects the level of mature factor D and / or pro-factor D in a biological sample obtained from a subject, thereby assessing the degree of APC activation in the subject. In some embodiments, the assays of the present invention can be used to assess the extent to which a complement pathway inhibitor (e.g., a MASP-3 inhibitor) reduces alternative complement pathway activation in vivo or in vitro. In some embodiments, the methods of the present invention are performed on a biological sample obtained from a subject. In some embodiments, the level of mature factor D and / or pro-factor D detected in the assays of the present invention is compared to a suitable reference value. The reference value may be, for example, a value measured from a sample obtained from a healthy patient (or a group of healthy patients), or a value measured from a sample obtained from a patient undergoing treatment with a MASP-3 inhibitor (e.g., obtained before treatment or at a certain time point during a course of treatment), or the reference value may be a predetermined threshold value. In one embodiment, the control sample is an individual or pooled sample from a subject suffering from an alternative pathway disease or disorder (e.g., paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, or other alternative pathway disease or disorder). In one embodiment, the control sample is an individual or pooled sample from a normal healthy volunteer. In one embodiment, the control sample is a baseline sample from a subject before treatment with a complement inhibitor (e.g., a MASP-3 inhibitor or other complement inhibitor). In one embodiment, the reference standard is at least one ratio of pro-factor D to mature factor D, mature factor D to total factor D, or pro-factor D to total factor D, which ratio is obtained from a test sample or a control sample (e.g., individual or pooled samples from normal healthy volunteers, or baseline samples from subjects before treatment with a complement inhibitor, or individual or pooled samples from subjects suffering from an alternative pathway disease or disorder).

[0224] As described herein, methods for detecting mature factor D and / or pro-factor D according to various aspects of the present disclosure can be used to assess the degree of alternative pathway complement activation, thereby used to define a pharmacodynamic endpoint or therapeutic threshold for complement inhibitors, such as alternative pathway complement inhibitors, such as MASP-3 inhibitors (e.g., MASP-3 inhibitory antibodies).

[0225] A. Methods for assessing the degree of alternative pathway complement activation in mammalian subjects In one aspect, the present disclosure provides a method for assessing the degree of alternative pathway complement (APC) activation in a test sample, performing an immunoassay comprising capturing and detecting maturation factor D in the test sample and / or capturing and detecting pro-factor D in the test sample, wherein the level of maturation factor D and / or the level of pro-factor D detected in the test sample is indicative of the degree of alternative pathway complement activation in the test sample. In one embodiment, the test sample is a biological sample obtained from a mammalian subject, and the method comprises assessing the degree of APC activation in the subject by performing an immunoassay comprising at least one of: (a) providing a biological sample obtained from the mammalian subject; and (b) capturing and detecting maturation factor D in the biological sample according to the methods of the invention described herein; and / or capturing and detecting pro-factor D in the biological sample. For example, in one embodiment, an immunoassay involves capturing and detecting mature factor D in a test sample, wherein the mature factor D is captured or detected with a mature factor D-specific monoclonal antibody, or a fragment thereof, that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but does not bind to pro-factor D. In one embodiment, an immunoassay involves capturing and detecting pro-factor D in a test sample, wherein the pro-factor D is captured or detected with a pro-factor D-specific monoclonal antibody, or a fragment thereof, that specifically binds to an epitope on the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but does not bind to mature factor D. In various embodiments, the method includes comparing the level of mature factor D detected in a test sample (e.g., a biological sample) with a predetermined level or a control sample, and / or comparing the level of pro-factor D detected in the test sample with a predetermined level or a control sample, and the level of mature factor D and / or pro-factor D detected in the test sample is indicative of the degree of alternative pathway complement activation in the test sample (e.g., a biological sample).In some embodiments, the method further includes using the results of the comparative analysis to provide diagnostic, prognostic, or treatment-related information about the mammalian subject from which the biological sample was obtained. In some embodiments, the present disclosure provides a method for evaluating the in vivo effect of a human complement inhibitor on alternative pathway complement activation. Any compound that binds to or otherwise blocks the production and / or activity of any of the human complement components may be utilized in accordance with the present disclosure. For example, the complement inhibitor can be, for example, a small molecule, a nucleic acid or nucleic acid analog, a peptidomimetic, or a macromolecule that is not a nucleic acid or protein, such as an antibody or fragment thereof. In some embodiments, the present disclosure provides a method for evaluating the in vivo effect of an inhibitor (e.g., an antibody or small molecule) specific for a human complement component, such as an inhibitor of a complement component selected from the group consisting of C1 (C1q, C1r, C1s), C2, C3, C4, C5, C6, C7, C8, C9, factor D, factor B, factor P, MBL, MASP-1, MASP-2, and MASP-3, on alternative complement pathway activation. In some embodiments, the present disclosure provides methods for evaluating the effect of an alternative complement pathway inhibitor on alternative pathway complement activation. In some embodiments, the present disclosure provides methods for evaluating the effect of a pro-factor D maturation inhibitor on alternative pathway complement activation.

[0226] In some embodiments, the present disclosure provides a method for evaluating the in vivo effect of a MASP-3 inhibitor administered to a mammalian subject on alternative pathway complement activation. In various embodiments, a MASP-3 inhibitor (e.g., a MASP-3 inhibitory antibody) is administered to a mammalian subject, and a biological sample is subsequently collected. The degree of alternative pathway complement (APC) activation in the biological sample is then evaluated by performing an immunoassay comprising at least one of capturing and detecting mature factor D in the biological sample according to the methods of the present invention described herein; and / or capturing and detecting pro-factor D in the biological sample.

[0227] B. Methods for Monitoring the Efficacy of MASP-3 Inhibitory Antibodies in Mammalian Subjects In one aspect, the present disclosure provides a method for monitoring the effectiveness of treatment with a MASP-3 inhibitory antibody in a mammalian subject, comprising: (a) administering a fixed dose of a MASP-3 inhibitory antibody to the mammalian subject at a first time point; (b) assessing a first concentration of maturation factor D and / or pro-factor D in a biological sample obtained from the subject after step (a); (c) treating the subject with the MASP-3 inhibitory antibody at a second time point; (d) assessing a second concentration of maturation factor D and / or pro-factor D in the biological sample obtained from the subject after step (c); and (e) comparing the level of maturation factor D and / or pro-factor D assessed in step (b) with the level of maturation factor D and / or pro-factor D assessed in step (d) to determine the effectiveness of the MASP-3 inhibitory antibody in the mammalian subject. In one aspect, the degree of APC activation in the subject is assessed by an immunoassay comprising capturing maturation factor D in the biological sample and detecting its level. Optionally, the level of maturation factor D detected in the biological sample is compared with a suitable reference value. The reference value can be, for example, the value of maturation factor D measured from a biological sample obtained from the subject before administration of the MASP-3 inhibitory antibody, an average value measured from samples obtained from a group of healthy control subjects, or a value representing the desired degree of APC activation (e.g., the level of maturation factor D corresponding to 90% inhibition of APC, or 80%, 70%, 60%, or 50% inhibition of APC). For example, a first biological sample is obtained from the subject before administration of the MASP-3 inhibitory antibody, and a second biological sample is obtained after administration of the MASP-3 inhibitory antibody, and the level of maturation factor D is measured in the sample. If the level of maturation factor D in the second biological sample is lower than the level of maturation factor D in the first biological sample or lower than a control value (e.g., a threshold value corresponding to the inhibition rate of APC), it can be concluded that the MASP-3 inhibitory antibody has inhibited APC activation to the desired degree.Alternatively, if the level of maturation factor D in the second biological sample is higher than the level of maturation factor D in the first biological sample or higher than a control value (e.g., a threshold corresponding to the inhibition rate of APC), it can be concluded that the dosage of the MASP-3 inhibitory antibody should be increased, and optionally, the method further comprises administering an increased dose of the MASP-3 inhibitory antibody to the subject.In some embodiments, when an increased dose of the MASP-3 inhibitory antibody is administered to the subject, steps (b) to (e) are repeated to determine whether the increased dose of the MASP-3 inhibitory antibody is sufficient to adjust the level of maturation factor D to the desired level compared to the respective control or reference standard.

[0228] In another embodiment, the degree of APC activation in a mammalian subject is assessed by an immunoassay comprising capturing pro-factor D in a biological sample and detecting its level. Optionally, the level of pro-factor D detected in the biological sample is compared with a suitable reference value. The reference value can be, for example, a value of pro-factor D measured from a biological sample obtained from the subject before administration of a MASP-3 inhibitory antibody, an average value measured from samples obtained from a group of healthy control subjects, or a value representing the desired degree of APC activation (e.g., a level of pro-factor D corresponding to 90% inhibition of APC, or 80%, or 70%, or 60%, or 50% inhibition of APC). For example, a first biological sample is obtained from the subject before administration of a MASP-3 inhibitory antibody, and a second biological sample is obtained after administration of the MASP-3 inhibitory antibody, and the level of pro-factor D is measured in the sample. If the level of pro-factor D in the second biological sample is greater than the level of pro-factor D in the first biological sample or higher than a control value (e.g., a threshold corresponding to the inhibition rate of APC), it can be concluded that the MASP-3 inhibitory antibody has inhibited APC activation to the desired extent. Alternatively, if the level of pro-factor D in the second biological sample is lower than the level of pro-factor D in the first biological sample or lower than a control value (e.g., a threshold corresponding to the inhibition rate of APC), it can be concluded that the dosage of the MASP-3 inhibitory antibody should be increased, and optionally, the method further comprises administering an increased dose of the MASP-3 inhibitory antibody to the subject. In some embodiments, if an increased dose of the MASP-3 inhibitory antibody is administered to the subject, steps (b) to (e) are repeated to determine whether the increased dose of the MASP-3 inhibitory antibody is sufficient to adjust the level of pro-factor D to the desired level compared to the respective control or reference standard.

[0229] In some embodiments, the methods are used to monitor the effectiveness of a MASP-3 inhibitory antibody administered to a human subject suffering from or at risk of developing an alternative pathway disease or disorder, e.g., alternative pathway diseases or disorders include paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD, and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microangiopathy, and the like. The disease is selected from the group consisting of immunological necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple sclerosis, Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft-versus-host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

[0230] C. Methods of Treating a Mammalian Subject Suffering from or at Risk for Developing an Alternative Pathway Disease or Disorder In another aspect, the present disclosure provides a method of treating a mammalian subject suffering from or at risk of developing an alternative pathway disease or disorder, comprising administering a MASP-3 inhibitory antibody to the subject when the subject is determined to have (i) a lower or decreased level of pro-factor D in one or more samples taken from the subject compared to a predetermined level of pro-factor D or compared to the level of pro-factor D in one or more control samples; and / or (ii) a higher or increased level of mature factor D in one or more samples taken from the subject compared to a predetermined level of mature factor D or compared to the level of mature factor D in one or more control samples. In one embodiment, the level of mature factor D in one or more samples taken from the subject is determined by performing an immunoassay involving the use of a mature factor D-specific monoclonal antibody. In one embodiment, the level of mature pro-factor D in one or more samples taken from the subject is determined by performing an immunoassay involving the use of a pro-factor D-specific monoclonal antibody.

[0231] In some embodiments, the methods are used to treat a human subject suffering from or at risk of developing an alternative pathway disease or disorder, e.g., the alternative pathway disease or disorder is paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD, and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescent formation, or the like. the disease is selected from the group consisting of chronic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple sclerosis, Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft-versus-host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

[0232] VIII. MASP-3 Inhibitors Human MASP-3 polypeptide (SEQ ID NO:7, from Genbank AAK84071.1) has 728 amino acid residues, including a 19-residue leader peptide. As noted above, MASP-3 is responsible for converting complement factor D from its proenzyme form (i.e., pro-factor D) to its mature form (i.e., mature factor D), demonstrating that the MASP-3 protein is therefore in an important upstream regulatory position for the alternative pathway. Accordingly, in implementing various aspects and embodiments of the present disclosure, representative MASP-3 inhibitors include agents that bind to or directly interact with MASP-3 shown as SEQ ID NO:7, and include anti-MASP-3 antibodies and MASP-3-binding fragments thereof, small molecules, and expression inhibitors that inhibit alternative pathway complement activation. In a preferred embodiment, the MASP-3 inhibitor is specific for MASP-3 and does not bind to MASP-1 or MASP-2. An example of a MASP-3 inhibitor is a MASP-3-specific inhibitor, e.g., a MASP-3 inhibitor that specifically binds to a portion of human MASP-3 (SEQ ID NO:7) with a binding affinity at least 10-fold greater than that of other components of the complement system. In one embodiment, the MASP-3 inhibitor is a high-affinity MASP-3 antibody that specifically binds to the serine protease domain of human MASP-3 (SEQ ID NO:7) with an affinity of less than 500 pM. In a preferred embodiment, the MASP-3 inhibitor, e.g., an antibody or its antigen-binding fragment or antigen-binding peptide, inhibits MASP-3-mediated maturation of factor D. MASP-3 inhibitors useful in the methods of the present invention can reduce MASP-3-dependent alternative pathway complement activation by more than 10%, e.g., more than 20%, more than 50%, or more than 90%. In one embodiment, the MASP-3 inhibitor reduces MASP-3-dependent alternative pathway complement activation by more than 90% (i.e., resulting in only 10% or less MASP-3 complement activation).

[0233] In one aspect, MASP-3 inhibitors useful in the methods of the present invention have high affinity (K<500 pM) for the serine protease domain of human MASP-3 (amino acid residues 450-728 of SEQ ID NO:7). DThe present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to the serine protease domain of MASP-3 and inhibits alternative pathway complement activation. For example, as described in WO2018 / 026722, which is incorporated herein by reference, and as further described in Example 10 and Tables 18-20 herein, numerous high-affinity anti-MASP-3 inhibitory antibodies have been produced that bind to the serine protease domain of MASP-3 and inhibit its catalytic activity. As further described in WO2018 / 026722, several representative MASP-3 inhibitory antibodies (e.g., 4D5, 10D12, and 13B1) have been humanized. Representative humanized MASP-3 inhibitory antibodies are described below.

[0234] Thus, in one aspect, a MASP-3 inhibitor for use in the claimed compositions and methods of the present invention comprises a monoclonal antibody that binds to a polypeptide consisting of human MASP-3 (SEQ ID NO:7), wherein the monoclonal antibody, or antigen-binding fragment thereof, binds to MASP-3 and comprises at least one of the following: (i) HC-CDR1 comprising SEQ ID NO:229 (TDDIN); a heavy chain variable region comprising HC-CDR2 comprising TIFF2025076475000047.tif4128, and HC-CDR3 comprising SEQ ID NO:236 (LEDTY); a light chain variable region comprising LC-CDR1 comprising TIFF2025076475000048.tif4128, LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and LC-CDR3 comprising SEQ ID NO: 242 (KQSYNLYT); (ii) HC-CDR1 comprising SEQ ID NO:230 (SYGMS); a heavy chain variable region comprising HC-CDR2 comprising TIFF2025076475000049.tif4128 and HC-CDR3 comprising SEQ ID NO:237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising TIFF2025076475000050.tif4128, an LC-CDR2 comprising SEQ ID NO:241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO:243 (WQGTHFPWT); or (iii) HC-CDR1 containing SEQ ID NO:231 (GKWIE); A heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES); and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT).

[0235] In one embodiment, the MASP-3 monoclonal antibody comprises a heavy chain variable region comprising at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identical to at least one of SEQ ID NO:220, SEQ ID NO:222, SEQ ID NO:223, SEQ ID NO:225, SEQ ID NO:226, or SEQ ID NO:228, and a light chain variable region comprising at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identical to at least one of SEQ ID NO:221, SEQ ID NO:224 or SEQ ID NO:227. In one embodiment, the MASP-3 monoclonal antibody comprises a heavy chain comprising at least 95% identical to SEQ ID NO:220 or SEQ ID NO:222 and a light chain comprising at least 95% identical to SEQ ID NO:221. In one embodiment, the MASP-3 monoclonal antibody comprises a heavy chain comprising at least 95% identity to SEQ ID NO:223 or SEQ ID NO:225 and a light chain comprising at least 95% identity to SEQ ID NO:224. In one embodiment, the MASP-3 monoclonal antibody comprises a heavy chain comprising at least 95% identity to SEQ ID NO:226 or SEQ ID NO:228 and a light chain comprising at least 95% identity to SEQ ID NO:227.

[0236] XIV. Pharmaceutical Compositions and Articles of Manufacture In another aspect, the present disclosure provides a pharmaceutical composition comprising a MASP-3 inhibitory antibody in an aqueous solution comprising a buffer system having pH 6.0±5%, 20±5% mM histidine, 100±5% mg / mL sucrose, and 0.035%±5% polysorbate 80, wherein the MASP-3 inhibitory antibody is present at a concentration of 110 mg / mL±5%, and the MASP-3 inhibitory antibody has an HC-CDR1 comprising SEQ ID NO:231 (GKWIE); TIFF2025076475000052.tif4128 or and a HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES); and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT). In one embodiment, the pharmaceutical composition is sterile. In one embodiment, the MASP-3 inhibitory antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identity to SEQ ID NO:226 or SEQ ID NO:227, and a light chain variable region comprising at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identity to SEQ ID NO:227. In one embodiment, the MASP-3 inhibitory antibody or antigen-binding fragment thereof is selected from the group consisting of a human antibody, a humanized antibody, a chimeric antibody, a murine antibody, and an antigen-binding fragment of any of the foregoing. In one embodiment, the MASP-3 inhibitory antibody or antigen-binding fragment thereof is selected from the group consisting of a single-chain antibody, an ScFv, a Fab fragment, a Fab' fragment, a F(ab')2 fragment, a monovalent antibody lacking a hinge region, and a whole antibody. In one embodiment, the MASP-3 inhibitory antibody further comprises an immunoglobulin constant region. In one embodiment, the MASP-3 inhibitory antibody comprises a human IgG4 constant region. In one embodiment, the MASP-3 inhibitory antibody comprises a human IgG4 constant region having an S228P mutation. In one embodiment, the MASP-3 inhibitory antibody comprises a mutation that promotes FcRn interaction at low pH, for example, the MASP-3 inhibitory antibody comprises the human IgG4 constant region shown as SEQ ID NO:245.

[0237] In one aspect, the present disclosure provides an article of manufacture containing a pharmaceutical composition comprising a MASP-3 inhibitory antibody in a unit dosage form suitable for therapeutic administration to a human subject, for example, a unit dosage in the range of 10 mg to 1000 mg (e.g., 50 mg to 800 mg, or 75 mg to 500, e.g., 100 mg to 300 mg, e.g., 125 to 275 mg, e.g., 150 to 200 mg, e.g., 150±5% mg, 155±5% mg, 160±5% mg, 165±5% mg, 170±5% mg, 175±5% mg, 180±5% mg, 185±5% mg or 190±5% mg), wherein the MASP-3 inhibitory antibody has an HC-CDR1 comprising SEQ ID NO:231 (GKWIE); TIFF2025076475000054.tif4128 or and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO:239, an LC-CDR2 comprising SEQ ID NO:178 (WASTRES); and an LC-CDR3 comprising SEQ ID NO:244 (KQSYNIPT).

[0238] In some embodiments, the article of manufacture comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, ampoules, pouches (e.g., intravenous infusion bags), vials, syringes, cartridges, etc. The container may be formed from a variety of materials, such as glass or plastic. The container holds a composition effective for treating a condition and may have a sterile access port (e.g., the container may be an intravenous solution bag or vial with a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a MASP-3 inhibitory antibody or antigen-binding fragment thereof of the present invention. The label or package insert indicates that the composition is used to treat a particular condition. The label or package insert further comprises instructions for administering the antibody composition to a patient.

[0239] In some embodiments, the pharmaceutical compositions and products described herein are for use in treating the subject suffering from or at risk of developing alternative pathway disease or disorder.In some embodiments, alternative pathway disease or disorder is paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP) or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, These are from the group consisting of Behçet's disease, multiple sclerosis, Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft-versus-host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

[0240] Exemplary Embodiments A. Maturation factor D-specific mAb: 1. An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in the N-terminal region of human maturation factor D, wherein the epitope comprises or consists of the amino acid ILGGREA (SEQ ID NO:5). 2. The isolated antibody or antigen-binding fragment thereof of paragraph 1, wherein the antibody specifically binds to human maturation factor D (SEQ ID NO:3) and does not bind to human pro-factor D (SEQ ID NO:2). 3. The isolated antibody or antigen-binding fragment thereof of paragraph 1 or 2, wherein the antibody is a monoclonal antibody. 4. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 3, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. 5. An isolated antibody or antigen-binding fragment thereof described in any of paragraphs 1 to 4, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. 6. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 4, wherein the antibody is a single-chain molecule. 7. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 4, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. 8. K<10nM D 8. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 7, which binds to human maturation factor D at 9. An isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 8, which is labeled with a detectable moiety. 10. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 9, which is immobilized on a substrate. 11. The isolated antibody or antigen-binding fragment thereof described in any of paragraphs 1 to 10, wherein the isolated antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, and the CDRs are numbered according to the Kabat numbering system. 12. The antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D comprises the following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO: 65), where X at position 1 is T, I, or S, and X at position 3 is G or I; (b) amino acid sequence HC-CDR2 containing TIFF2025076475000056.tif4128 (wherein X at position 11 is H or N and X at position 16 is S or R); (c) amino acid sequence HC-CDR3 containing TIFF2025076475000057.tif4128 (in which X at position 6 is R, G, or N, X at position 7 is S or Y, X at position 8 is F, I, or V, and X at position 10 is D or H); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69) (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54). 11. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 10, comprising a binding domain comprising: 13. The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:25, (b) HC-CDR2 comprising SEQ ID NO:27, (c) HC-CDR3 comprising SEQ ID NO:29, (d) LC-CDR1 comprising SEQ ID NO:50, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. 13. The isolated antibody or antigen-binding fragment thereof of paragraph 12, comprising: 14. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:12 or SEQ ID NO:13; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:19; (c) a VH comprising SEQ ID NO: 12 and a VL comprising SEQ ID NO: 18; and / or (d) a VH domain comprising SEQ ID NO:13 and a VL domain comprising SEQ ID NO:19 14. The isolated antibody or antigen-binding fragment thereof of paragraph 13, comprising at least one of: 15. The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 33, (b) HC-CDR2 comprising SEQ ID NO: 34, (c) HC-CDR3 comprising SEQ ID NO: 36, (d) LC-CDR1 comprising SEQ ID NO: 58, (e) LC-CDR2 comprising SEQ ID NO: 52, and (f) LC-CDR3 comprising SEQ ID NO: 54. 13. The isolated antibody or antigen-binding fragment thereof of paragraph 12, comprising: 16. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:14; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:20; and / or (c) a VH domain comprising SEQ ID NO:14 and a VL domain comprising SEQ ID NO:20 16. The isolated antibody or antigen-binding fragment thereof of paragraph 15, comprising at least one of: 17. The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:38, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:41, (d) LC-CDR1 comprising SEQ ID NO:60, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. 13. The isolated antibody or antigen-binding fragment thereof of paragraph 12, comprising: 18. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:15; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:21; and / or (c) a VH domain comprising SEQ ID NO:15 and a VL domain comprising SEQ ID NO:21 18. The isolated antibody or antigen-binding fragment thereof of paragraph 17, comprising at least one of: 19. The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:43, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:41, (d) LC-CDR1 comprising SEQ ID NO:62, (e) LC-CDR22 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:54. 13. The isolated antibody or antigen-binding fragment thereof of paragraph 12, comprising: 20. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:16; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:22; and / or (c) a VH domain comprising SEQ ID NO: 16 and a VL domain comprising SEQ ID NO: 22 20. The isolated antibody or antigen-binding fragment thereof of paragraph 19, comprising at least one of: 21. The binding domain comprises the following six CDRs: (a) HC-CDR1 comprising SEQ ID NO:43, (b) HC-CDR2 comprising SEQ ID NO:39, (c) HC-CDR3 comprising SEQ ID NO:47, (d) LC-CDR1 comprising SEQ ID NO:63, (e) LC-CDR2 comprising SEQ ID NO:64, and (f) LC-CDR3 comprising SEQ ID NO:54. 13. The isolated antibody or antigen-binding fragment thereof of paragraph 12, comprising: 22. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:17; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:23; and / or (c) a VH domain comprising SEQ ID NO:17 and a VL domain comprising SEQ ID NO:23 22. The isolated antibody or antigen-binding fragment thereof of paragraph 21, comprising at least one of: 23. A nucleic acid molecule encoding the amino acid sequence of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to any one of paragraphs 11 to 22. 24. An expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of the present invention described in paragraph 23. 25. A cell comprising at least one nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D of the present invention according to paragraph 23 or paragraph 24. 26. A method for producing an isolated antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, comprising culturing the cell described in paragraph 25 under conditions that allow expression of a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, and isolating the anti-maturation factor D-specific antibody or antigen-binding fragment thereof. 27. A composition comprising an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to any of paragraphs 1 to 22. 28. A substrate for use in an immunoassay comprising at least one antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to any of paragraphs 1-22. 29. A kit for detecting the presence or amount of maturation factor D in a test sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D described in any of paragraphs 1-22.

[0241] B. Pro-factor D specific mAb: 1. An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human factor D, wherein the epitope comprises or consists of "APPRGR" (SEQ ID NO:4). 2. The antibody or antigen-binding fragment thereof described in paragraph 1, which specifically binds to human pro-factor D (SEQ ID NO:2) and does not bind to mature factor D (SEQ ID NO:3). 3. The isolated antibody or antigen-binding fragment thereof of paragraph 1 or 2, wherein the antibody is a monoclonal antibody. 4. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 3, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. 5. An isolated antibody or antigen-binding fragment thereof described in any of paragraphs 1 to 4, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. 6. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 4, wherein the antibody is a single-chain molecule. 7. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 4, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. 8. K<10nM D 8. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 7, which binds to human profactor D at 9. An isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 8, which is labeled with a detectable moiety. 10. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 9, which is immobilized on a substrate. 11. An isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D described in any of paragraphs 1 to 10, wherein the antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, and the CDRs are numbered according to the Kabat numbering system. 12. An isolated antibody or antigen-binding fragment thereof described in paragraph 11, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 139, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 145. 13. The following six CDRs: (a) HC-CDR1 comprising the amino acid sequence XYWMS (SEQ ID NO:201), where X at position 1 is N, S, or T; (b) the amino acid sequence (c) HC-CDR2 comprising the amino acid sequence AWFAX (SEQ ID NO: 203) (wherein X at position 5 is S, Y, or N); (d) amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence WASTRES (SEQ ID NO: 178); and (f) an LC-CDR3 comprising the amino acid sequence LQYYXYPYT (SEQ ID NO: 205) (wherein X at position 5 is T or S). 13. The isolated antibody or antigen-binding fragment thereof of paragraph 12, comprising a binding domain comprising: 14. The following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 149 or SEQ ID NO: 155, (b) HC-CDR2 comprising SEQ ID NO: 151 or SEQ ID NO: 156, (c) HC-CDR3 comprising SEQ ID NO: 153, (d) LC-CDR1 comprising SEQ ID NO: 176, (e) LC-CDR2 comprising SEQ ID NO: 178, and (f) LC-CDR3 comprising SEQ ID NO: 180. 14. The isolated antibody or antigen-binding fragment thereof of paragraph 13, comprising a binding domain comprising: 15. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:136; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:137; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:142; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:143; (e) a VH domain comprising SEQ ID NO: 136 and a VL domain comprising SEQ ID NO: 142; and / or (f) a VH domain comprising SEQ ID NO: 137 and a VL domain comprising SEQ ID NO: 143 15. The isolated antibody or antigen-binding fragment thereof of paragraph 14, comprising at least one of: 16. The following six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 158, (b) HC-CDR2 comprising SEQ ID NO: 159 or SEQ ID NO: 163, (c) HC-CDR3 comprising SEQ ID NO: 161 or SEQ ID NO: 165, (d) LC-CDR1 comprising SEQ ID NO: 184 or SEQ ID NO: 189, (e) LC-CDR2 comprising SEQ ID NO: 178, and (f) LC-CDR3 comprising SEQ ID NO: 187. 14. The isolated antibody or antigen-binding fragment thereof of paragraph 13, comprising a binding domain comprising: 17. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:138; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:139; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:144; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:145; (e) a VH domain comprising SEQ ID NO: 138 and a VL domain comprising SEQ ID NO: 144; and / or (f) a VH domain comprising SEQ ID NO: 139 and a VL domain comprising SEQ ID NO: 145 17. The isolated antibody or antigen-binding fragment thereof of paragraph 16, comprising at least one of: 18. An isolated antibody or antigen-binding fragment thereof described in paragraph 11, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NO:140 and SEQ ID NO:141, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO:146 and SEQ ID NO:147. 19. The following six CDRs: (a) CDR-H1 comprising SEQ ID NO: 167, (b) CDR-H2 comprising SEQ ID NO: 169 or SEQ ID NO: 173, (c) CDR-H3 comprising SEQ ID NO: 171 or SEQ ID NO: 174, (d) CDR-L1 comprising SEQ ID NO: 194, (e) CDR-L2 comprising SEQ ID NO: 196 or SEQ ID NO: 199, and (f) CDR-L3 comprising SEQ ID NO: 198 or SEQ ID NO: 200. 19. The isolated antibody or antigen-binding fragment thereof of paragraph 18, comprising a binding domain comprising: 20. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:140; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:141; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:146; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:147; (e) a VH domain comprising SEQ ID NO:140 and a VL domain comprising SEQ ID NO:146; and / or (f) a VH domain comprising SEQ ID NO: 141 and a VL domain comprising SEQ ID NO: 147 20. The isolated antibody or antigen-binding fragment thereof of paragraph 19, comprising at least one of: 21. A nucleic acid molecule encoding the amino acid sequence of an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any of paragraphs 11 to 20. 22. An expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to paragraph 21. 23. A cell comprising at least one nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to paragraph 21 or paragraph 22. 24. A method for producing an isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D, comprising culturing the cell of paragraph 23 under conditions that allow expression of a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human profactor D, and isolating the anti-profactor D-specific antibody or antigen-binding fragment thereof. 25. A composition comprising an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any of paragraphs 1 to 20. 26. A substrate for use in an immunoassay comprising at least one antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any of paragraphs 1-20. 27. A kit for detecting the presence or amount of pro-factor D in a test sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that specifically binds to human mature factor D described in any of paragraphs 1-20.

[0242] C. Anti-Factor D mAb (detects pro- and mature factor D by binding to a common epitope) 1. An isolated antibody or antigen-binding fragment thereof that binds to an epitope common to both human maturation factor D and human pro-factor D, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85 to 88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89 to 93, wherein the CDRs are numbered according to the Kabat numbering system. 2. The isolated antibody or antigen-binding fragment thereof of paragraph 1, wherein the antibody is a monoclonal antibody. 3. The isolated antibody or antigen-binding fragment thereof of paragraph 1 or 2, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody. 4. An isolated antibody or antigen-binding fragment thereof described in any of paragraphs 1 to 3, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2. 5. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 4, which is a single-chain molecule. 6. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 4, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4. 7. K<10nM D 8. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 7, which binds to human factor D at 8. An isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 7, which is labeled with a detectable moiety. 9. An isolated antibody or antigen-binding fragment thereof according to any of paragraphs 1 to 8, which is immobilized on a substrate. 10. An isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D described in any of paragraphs 1 to 9, wherein the antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, and the CDRs are numbered according to the Kabat numbering system. 11. The following six CDRs: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO:95; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO:97; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO:99; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO:111; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO:113; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO:115. 11. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 10, comprising a binding domain comprising: 12. The following six CDRs: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO: 101; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO: 103 or 107; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO: 105 or 108; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO: 60 or 123; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO: 119, 124, or 126; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO: 121 or 125. 11. The isolated antibody or antigen-binding fragment thereof of any of paragraphs 1 to 10, comprising a binding domain comprising: 13. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:85; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:86; (c) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:87; (d) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:88; (e) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:89; (f) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:90; (g) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:91; (h) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:92; (i) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:93; (j) a VH domain comprising SEQ ID NO:85 and a VL domain comprising SEQ ID NO:89 or SEQ ID NO:90; (k) a VH domain comprising SEQ ID NO:86 and a VL domain comprising SEQ ID NO:91; (l) a VH domain comprising SEQ ID NO:87 and a VL domain comprising SEQ ID NO:92; and / or (m) a VH domain comprising SEQ ID NO:88 and a VL domain comprising SEQ ID NO:93 11. The isolated antibody or antigen-binding fragment thereof of paragraph 10, comprising at least one of: 14. A nucleic acid molecule encoding the amino acid sequence of an antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D described in any of paragraphs 10 to 13. 15. An expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D according to paragraph 14. 16. A cell comprising at least one of the nucleic acid molecules encoding an antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D described in paragraph 14 or paragraph 15. 17. A method for producing an isolated antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, comprising culturing the cell described in paragraph 16 under conditions that allow expression of a nucleic acid molecule encoding the antibody or antigen-binding fragment thereof that binds to human factor D, and isolating the anti-factor D antibody or antigen-binding fragment thereof. 18. A composition comprising an antibody or antigen-binding fragment thereof that specifically binds to an epitope common to both human mature factor D and human pro-factor D described in any of paragraphs 1 to 13. 19. A substrate for use in an immunoassay comprising at least one antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D according to any of paragraphs 1-13. 20. A kit for detecting the presence of factor D in a biological sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D described in any of paragraphs 1-13.

[0243] D. Kits for detecting mature factor D and / or pro-factor D in immunoassays 1. A kit comprising at least one monoclonal antibody or antigen-binding fragment thereof that specifically detects or quantifies human maturation factor D (SEQ ID NO:3) and / or pro-factor D (SEQ ID NO:2) in an immunoassay, wherein the at least one monoclonal antibody or antigen-binding fragment thereof is: (i) a monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope encompassing the amino terminus of human maturation factor D, wherein the epitope comprises or consists of the amino acid sequence ILGGREA (SEQ ID NO:5), and the antibody does not bind to human profactor D (SEQ ID NO:2); or (ii) A monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human factor D, wherein the epitope comprises or consists of "APPRGR" (SEQ ID NO:4), and the antibody does not bind to mature factor D (SEQ ID NO:3). Includes a kit. 2. The kit of paragraph 1, further comprising an antibody or fragment thereof that binds to an epitope common to both human maturation factor D (SEQ ID NO:3) and human pro-factor D (SEQ ID NO:2). 3. The kit of paragraph 1 or 2, further comprising at least one container. 4. The kit of any of paragraphs 1-3, wherein the antibody or antigen-binding fragment of paragraph 1 subpart (i) that specifically binds to human maturation factor D comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12-17, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18-23, the CDRs being numbered according to the Kabat numbering system. 5. The kit of any of paragraphs 1 to 3, wherein the antibody or antigen-binding fragment of paragraph 1 subpart (ii) that specifically binds to an epitope in the activation ("pro") peptide of human factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141 and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, the CDRs being numbered according to the Kabat numbering system. 6. The kit of any of paragraphs 1 to 5, wherein the immunoassay is an enzyme-linked immunosorbent assay (ELISA). 7. The kit of any of paragraphs 1 to 6, wherein the antibody or antigen-binding fragment thereof of paragraph 1 subpart (i) is a coating antibody. 8. The kit of any of paragraphs 1 to 6, wherein the antibody or antigen-binding fragment thereof of paragraph 1 subpart (i) is a detection antibody. 9. The kit of any of paragraphs 1 to 6, wherein the antibody or antigen-binding fragment thereof of paragraph 1 subpart (ii) is a coating antibody. 10. The kit of any of paragraphs 1 to 6, wherein the antibody or antigen-binding fragment thereof of paragraph 1 subpart (ii) is a detection antibody. 11. The kit of any of paragraphs 1 to 10, further comprising an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D (SEQ ID NO:3) and human pro-factor D (SEQ ID NO:2). 12. The kit described in paragraph 11, wherein the anti-factor D antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85 to 88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89 to 93, and the CDRs are numbered according to the Kabat numbering system.

[0244] E. Assay for detecting maturation factor D 1. A method for determining the presence or amount of mature factor D in a test sample, comprising: (a) contacting a test sample with an anti-human maturation factor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence, absence or amount of said antibody or antigen-binding fragment thereof bound to maturation factor D, wherein the presence of said binding indicates the presence or amount of maturation factor D in said sample. Including, The method, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof binds to an epitope in the N-terminal region of maturation factor D represented by amino acids ILGGREA (SEQ ID NO:5). 2. The method of paragraph 1, wherein the antibody or antigen-binding fragment thereof specifically binds to human mature factor D (SEQ ID NO:3) and does not bind...

Claims

1. An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in the amino-terminal region of human maturation factor D, wherein the epitope comprises or consists of the amino acid sequence ILGGREA (SEQ ID NO:5).

2. 2. The isolated antibody or antigen-binding fragment thereof of claim 1, which specifically binds to human maturation factor D (SEQ ID NO: 3) and does not bind to human pro-factor D (SEQ ID NO: 2).

3. 3. The isolated antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody is a monoclonal antibody.

4. The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody.

5. The antigen-binding fragment is an Fv, Fab, Fab', F(ab) 2 and F(ab') 2 The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, selected from the group consisting of:

6. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 5, which is a single chain molecule.

7. The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4.

8. K less than 10 nM D The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, which binds to human maturation factor D at

9. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 8, which is labeled with a detectable moiety.

10. The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, which is immobilized on a substrate.

11. The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, wherein the isolated antibody or antigen-binding fragment thereof specifically binds to human maturation factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, wherein the CDRs are numbered according to the Kabat numbering system.

12. The antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D has six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO: 65), in which X at position 1 is T, I or S, and X at position 3 is G or I; (b) the amino acid sequence (c) an amino acid sequence comprising: (wherein X at position 6 is R, G or N, X at position 7 is S or Y, X at position 8 is F, I or V, and X at position 10 is D or H); (d) an amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69), (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54).

12. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 11, comprising a binding domain comprising:

13. The binding domain comprises six CDRs: (a) HC-CDR1 comprising SEQ ID NO:25, (b) HC-CDR2 comprising SEQ ID NO:27, (c) HC-CDR3 comprising SEQ ID NO:29, (d) LC-CDR1 comprising SEQ ID NO:50, (e) LC-CDR2 comprising SEQ ID NO:52, and (f) LC-CDR3 comprising SEQ ID NO:

54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 12, comprising:

14. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:12 or SEQ ID NO:13; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:19; (c) a VH comprising SEQ ID NO: 12 and a VL comprising SEQ ID NO: 18; and / or (d) a VH domain comprising SEQ ID NO: 13 and a VL domain comprising SEQ ID NO: 19 14. The isolated antibody or antigen-binding fragment thereof of claim 13, comprising at least one of the following:

15. The binding domain comprises six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 33, (b) HC-CDR2 comprising SEQ ID NO: 34, (c) HC-CDR3 comprising SEQ ID NO: 36, (d) LC-CDR1 comprising SEQ ID NO: 58, (e) LC-CDR2 comprising SEQ ID NO: 52, and (f) LC-CDR3 comprising SEQ ID NO:

54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 12, comprising:

16. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 14; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:20; and / or (c) a VH domain comprising SEQ ID NO: 14 and a VL domain comprising SEQ ID NO: 20 16. The isolated antibody or antigen-binding fragment thereof of claim 15, comprising at least one of the following:

17. The binding domain comprises six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 38, (b) HC-CDR2 comprising SEQ ID NO: 39, (c) HC-CDR3 comprising SEQ ID NO: 41, (d) LC-CDR1 comprising SEQ ID NO: 60, (e) LC-CDR2 comprising SEQ ID NO: 52, and (f) LC-CDR3 comprising SEQ ID NO:

54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 12, comprising:

18. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:15; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 21; and / or (c) a VH domain comprising SEQ ID NO: 15 and a VL domain comprising SEQ ID NO: 21 20. The isolated antibody or antigen-binding fragment thereof of claim 17, comprising at least one of the following:

19. The binding domain comprises six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 43, (b) HC-CDR2 comprising SEQ ID NO: 39, (c) HC-CDR3 comprising SEQ ID NO: 41, (d) LC-CDR1 comprising SEQ ID NO: 62, (e) LC-CDR22 comprising SEQ ID NO: 52, and (f) LC-CDR3 comprising SEQ ID NO:

54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 12, comprising:

20. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:16; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 22; and / or (c) a VH domain comprising SEQ ID NO: 16 and a VL domain comprising SEQ ID NO: 22 20. The isolated antibody or antigen-binding fragment thereof of claim 19, comprising at least one of the following:

21. The binding domain comprises six CDRs: (a) an HC-CDR1 comprising SEQ ID NO: 43; (b) an HC-CDR2 comprising SEQ ID NO: 39; (c) an HC-CDR3 comprising SEQ ID NO: 47; (d) an LC-CDR1 comprising SEQ ID NO: 63; (e) an LC-CDR2 comprising SEQ ID NO: 64; and (f) an LC-CDR3 comprising SEQ ID NO:

54. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 12, comprising:

22. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 17; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 23; and / or (c) a VH domain comprising SEQ ID NO: 17 and a VL domain comprising SEQ ID NO: 23 22. The isolated antibody or antigen-binding fragment thereof of claim 21, comprising at least one of the following:

23. A nucleic acid molecule encoding the CDRs of the heavy chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to any one of claims 11 to 22.

24. A nucleic acid molecule encoding the CDRs of the light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to any one of claims 11 to 22.

25. A cloning vector or expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to claim 23 and / or claim 24.

26. A cell comprising at least one of the nucleic acid molecules encoding an antibody or an antigen-binding fragment thereof that specifically binds to human maturation factor D according to claim 23 or claim 24.

27. A method for producing an isolated antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, comprising culturing a cell described in claim 26 under conditions that allow expression of a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D, and isolating an anti-maturation factor D specific antibody or antigen-binding fragment thereof.

28. A composition comprising an antibody or an antigen-binding fragment thereof that specifically binds to human maturation factor D according to any one of claims 1 to 22.

29. A substrate for use in an immunoassay comprising at least one antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D according to any one of claims 1 to 22.

30. A kit for detecting the presence or amount of maturation factor D in a test sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that specifically binds to human maturation factor D described in any one of claims 1 to 22.

31. An isolated antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human factor D, wherein the epitope comprises or consists of "APPRGR" (SEQ ID NO:4).

32. 32. The isolated antibody or antigen-binding fragment thereof of claim 31, which specifically binds to human pro-factor D (SEQ ID NO: 2) and does not bind to mature factor D (SEQ ID NO: 3).

33. 33. The isolated antibody or antigen-binding fragment thereof of claim 31 or 32, wherein the antibody is a monoclonal antibody.

34. The isolated antibody or antigen-binding fragment thereof according to any one of claims 31 to 33, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody.

35. The antigen-binding fragment is an Fv, Fab, Fab', F(ab) 2 and F(ab') 2 35. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 34, selected from the group consisting of:

36. 36. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 35, which is a single chain molecule.

37. 37. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 36, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4.

38. K less than 10 nM D 38. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 37, which binds to human profactor D at

39. 39. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 38, which is labeled with a detectable moiety.

40. 40. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 39, which is immobilized on a substrate.

41. 40. An isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any one of claims 31 to 39, wherein the antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, wherein the CDRs are numbered according to the Kabat numbering system.

42. 42. An isolated antibody or antigen-binding fragment thereof according to any one of claims 31 to 41, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 139, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 145.

43. The six CDRs are: (a) HC-CDR1, which comprises the amino acid sequence XYWMS (SEQ ID NO:201), where X at position 1 is N, S or T; (c) an HC-CDR3 comprising the amino acid sequence AWFAX (SEQ ID NO:203), in which X at position 5 is S, Y or N; (d) an amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence WASTRES (SEQ ID NO: 178); and (f) an LC-CDR3 comprising the amino acid sequence LQYYXYPYT (SEQ ID NO: 205), wherein X at position 5 is T or S.

43. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 42, comprising a binding domain comprising:

44. The six CDRs are: (a) HC-CDR1 comprising SEQ ID NO: 149 or SEQ ID NO: 155; (b) HC-CDR2 comprising SEQ ID NO: 151 or SEQ ID NO: 156; (c) HC-CDR3 comprising SEQ ID NO: 153; (d) LC-CDR1 comprising SEQ ID NO: 176; (e) LC-CDR2 comprising SEQ ID NO: 178; and (f) LC-CDR3 comprising SEQ ID NO:

180.

44. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 43, comprising a binding domain comprising:

45. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:136; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:137; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:142; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 143; (e) a VH domain comprising SEQ ID NO: 136 and a VL domain comprising SEQ ID NO: 142; and / or (f) a VH domain comprising SEQ ID NO: 137 and a VL domain comprising SEQ ID NO: 143 45. The isolated antibody or antigen-binding fragment thereof of claim 44, comprising at least one of the following:

46. The six CDRs are: (a) HC-CDR1 comprising SEQ ID NO: 158; (b) HC-CDR2 comprising SEQ ID NO: 159 or SEQ ID NO: 163; (c) HC-CDR3 comprising SEQ ID NO: 161 or SEQ ID NO: 165; (d) LC-CDR-1 comprising SEQ ID NO: 184 or SEQ ID NO: 189; (e) LC-CDR2 comprising SEQ ID NO: 178; and (f) LC-CDR3 comprising SEQ ID NO:

187.

44. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 43, comprising a binding domain comprising:

47. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 138; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 139; (b) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 144; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:145; (d) a VH domain comprising SEQ ID NO: 138 and a VL domain comprising SEQ ID NO: 144; and / or (e) a VH domain comprising SEQ ID NO: 139 and a VL domain comprising SEQ ID NO: 145 47. The isolated antibody or antigen-binding fragment thereof of claim 46, comprising at least one of the following:

48. 42. The isolated antibody or antigen-binding fragment thereof according to any one of claims 31 to 41, comprising a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NO: 140 and SEQ ID NO: 141, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO: 146 and SEQ ID NO:

147.

49. The six CDRs are: (a) a CDR-H1 comprising SEQ ID NO:167; (b) a CDR-H2 comprising SEQ ID NO:169 or SEQ ID NO:173; (c) a CDR-H3 comprising SEQ ID NO:171 or SEQ ID NO:174; (d) a CDR-L1 comprising SEQ ID NO:194; (e) a CDR-L2 comprising SEQ ID NO:196 or SEQ ID NO:199; and (f) a CDR-L3 comprising SEQ ID NO:198 or SEQ ID NO:

200.

42. The isolated antibody or antigen-binding fragment thereof of any one of claims 31 to 41, comprising a binding domain comprising:

50. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:140; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:141; (c) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:146; (d) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:147; (e) a VH domain comprising SEQ ID NO: 140 and a VL domain comprising SEQ ID NO: 146; and / or (f) a VH domain comprising SEQ ID NO: 141 and a VL domain comprising SEQ ID NO: 147 50. The isolated antibody or antigen-binding fragment thereof of claim 49, comprising at least one of the following:

51. A nucleic acid molecule encoding the CDRs of the heavy chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any one of claims 31 to 50.

52. A nucleic acid molecule encoding the CDRs of the light chain variable region of an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any one of claims 31 to 50.

53. 53. A cloning vector or expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to claim 51 and / or claim 52.

54. A cell comprising at least one of the nucleic acid molecules encoding an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to claim 51 and / or claim 52.

55. A method for producing an isolated antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, comprising culturing the cell of claim 54 under conditions that allow expression of a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to human pro-factor D, and isolating an anti-pro-factor D specific antibody or antigen-binding fragment thereof.

56. A composition comprising an antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any one of claims 31 to 50.

57. A substrate for use in an immunoassay comprising at least one antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any one of claims 31 to 50.

58. A kit for detecting the presence or amount of pro-factor D in a test sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that specifically binds to human mature factor D described in any one of claims 31 to 50.

59. An isolated antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D, wherein the antibody comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85-88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89-93, wherein the CDRs are numbered according to the Kabat numbering system.

60. 60. The isolated antibody or antigen-binding fragment thereof of claim 59, wherein the antibody is a monoclonal antibody.

61. 61. The isolated antibody or antigen-binding fragment thereof of claim 59 or 60, wherein the antibody is a humanized antibody, a chimeric antibody, or a fully human antibody.

62. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 61, wherein the antigen-binding fragment is selected from the group consisting of Fv, Fab, Fab', F(ab)2, and F(ab')2.

63. 63. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 62, wherein the antibody is a single chain molecule.

64. 64. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 63, wherein the antibody is an IgG molecule selected from the group consisting of IgG1, IgG2, and IgG4.

65. K less than 10 nM D 65. The isolated antibody or antigen-binding fragment thereof according to any one of claims 59 to 64, which binds to human factor D at

66. 66. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 65, which is labeled with a detectable moiety.

67. 67. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 66, which is immobilized on a substrate.

68. 68. An isolated antibody or antigen-binding fragment thereof that specifically binds to human profactor D according to any one of claims 59 to 67, wherein the antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, wherein the CDRs are numbered according to the Kabat numbering system.

69. The six CDRs are: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO:95; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO:97; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO:99; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO:111; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO:113; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO:

115.

69. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 68, comprising a binding domain comprising:

70. The six CDRs are: (a) HC-CDR1 comprising the amino acid sequence SEQ ID NO: 101; (b) HC-CDR2 comprising the amino acid sequence SEQ ID NO: 103 or 107; (c) HC-CDR3 comprising the amino acid sequence SEQ ID NO: 105 or 108; (d) LC-CDR1 comprising the amino acid sequence SEQ ID NO: 60 or 123; (e) LC-CDR2 comprising the amino acid sequence SEQ ID NO: 119, 124, or 126; and (f) LC-CDR3 comprising the amino acid sequence SEQ ID NO: 121 or 125.

69. The isolated antibody or antigen-binding fragment thereof of any one of claims 59 to 68, comprising a binding domain comprising:

71. (a) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:85; (b) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 86; (c) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 87; (d) a VH domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 88; (e) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 89; (f) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:90; (g) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:91; (h) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:92; (i) a VL domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:93; (j) a VH domain comprising SEQ ID NO: 85 and a VL domain comprising SEQ ID NO: 89 or SEQ ID NO: 90; (k) a VH domain comprising SEQ ID NO: 86 and a VL domain comprising SEQ ID NO: 91; (l) a VH domain comprising SEQ ID NO: 87 and a VL domain comprising SEQ ID NO: 92; and / or (m) a VH domain comprising SEQ ID NO: 88 and a VL domain comprising SEQ ID NO: 93 71. The isolated antibody or antigen-binding fragment thereof of claim 70, comprising at least one of the following:

72. A nucleic acid molecule encoding the CDRs of the heavy chain variable region of an antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D according to any one of claims 59 to 71.

73. A nucleic acid molecule encoding the CDRs of the light chain variable region of an antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D according to any one of claims 59 to 71.

74. A cloning vector or expression cassette comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D according to claim 72 and / or claim 73.

75. A cell comprising at least one of the nucleic acid molecules encoding an antibody or an antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D described in claim 72 and / or claim 73.

76. A method for producing an isolated antibody or antigen-binding fragment thereof that binds to an epitope common to human mature factor D and human pro-factor D, comprising culturing the cell of claim 75 under conditions that allow expression of a nucleic acid molecule encoding the antibody or antigen-binding fragment thereof that binds to factor D, and isolating the anti-factor D specific antibody or antigen-binding fragment thereof.

77. A composition comprising an antibody or an antigen-binding fragment thereof that specifically binds to an epitope common to human maturation factor D and human profactor D according to any one of claims 59 to 71.

78. A substrate for use in an immunoassay comprising at least one antibody or antigen-binding fragment thereof that binds to an epitope common to human maturation factor D and human profactor D according to any one of claims 59 to 71.

79. A kit for detecting the presence or amount of factor D in a test sample, comprising: (a) at least one container; and (b) at least one antibody or antigen-binding fragment thereof that binds to an epitope common to human mature factor D and human pro-factor D described in any one of claims 59 to 71.

80. A kit comprising at least one monoclonal antibody that specifically detects or quantifies human maturation factor D (SEQ ID NO:3) and / or pro-factor D (SEQ ID NO:2) in an immunoassay, the at least one monoclonal antibody being (i) a maturation factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope that encompasses the amino terminus of human maturation factor D, wherein the epitope comprises or consists of the amino acids ILGGREA (SEQ ID NO:5), and the antibody does not bind to human profactor D (SEQ ID NO:2); and / or (ii) a pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide of human factor D, wherein the epitope comprises or consists of "APPRGR" (SEQ ID NO:4), and the antibody does not bind to mature factor D (SEQ ID NO:3); Including the kit.

81. The kit of claim 80, further comprising an anti-factor D antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D (SEQ ID NO:3) and human pro-factor D (SEQ ID NO:2).

82. 82. The kit of claim 80 or 81, further comprising at least one container.

83. The kit according to any one of claims 80 to 82, wherein the antibody or antigen-binding fragment thereof of subpart (i) specifically binds to human maturation factor D comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, wherein the CDRs are numbered according to the Kabat numbering system.

84. The kit of any one of claims 80 to 83, wherein the antibody or antigen-binding fragment thereof of subpart (ii) that specifically binds to an epitope in the activation ("pro") peptide of human factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141 and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, wherein the CDRs are numbered according to the Kabat numbering system.

85. The kit of any one of claims 80 to 84, wherein the immunoassay is an enzyme-linked immunosorbent assay (ELISA).

86. 81. The kit of claim 80, wherein the mature factor D-specific antibody or antigen-binding fragment thereof of subpart (i) is a coating antibody.

87. 81. The kit of claim 80, wherein the mature factor D-specific antibody or antigen-binding fragment thereof of subpart (i) is a detection antibody.

88. The kit of claim 80, wherein the pro-factor D specific antibody or antigen-binding fragment thereof of subpart (ii) is a coating antibody.

89. The kit of claim 80, wherein the pro-factor D specific antibody or antigen-binding fragment thereof of subpart (ii) is a detection antibody.

90. The kit of claim 81, wherein the anti-factor D antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 85-88, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 89-93, wherein the CDRs are numbered according to the Kabat numbering system.

91. 1. A method for determining the presence or amount of mature factor D in a test sample, comprising: (a) contacting a test sample with a mature factor D-specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or absence or amount of said antibody or antigen-binding fragment thereof bound to maturation factor D, wherein the presence of said binding indicates the presence or amount of maturation factor D in the sample. Including, The method, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof binds to an epitope in the N-terminal region of maturation factor D represented by amino acids ILGGREA (SEQ ID NO:5).

92. The method of claim 91, wherein the antibody or antigen-binding fragment thereof specifically binds to human mature factor D (SEQ ID NO:3) and does not bind to human pro-factor D (SEQ ID NO:2).

93. 93. The method of claim 91 or 92, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof is immobilized on a substrate.

94. The method of any one of claims 91 to 93, wherein the immunoassay is an ELISA assay.

95. The method of any one of claims 91 to 94, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof is labeled with a detectable moiety, and step (b) comprises detecting the presence or amount of the detectable moiety.

96. The method of any one of claims 91 to 94, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof is naked (i.e., unlabeled) and the presence or amount of the antibody or fragment thereof bound to maturation factor D is detected using a labeled antibody that binds to the anti-maturation factor D antibody.

97. The method of any one of claims 91 to 96, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof is immobilized (i.e., captured / coated) on a substrate, and the bound maturation factor D is detected using a second antibody that binds to a different epitope of factor D.

98. 98. The method of any one of claims 91-97, wherein the test sample is a biological sample obtained from a mammalian subject, e.g., the biological sample is selected from the group consisting of blood, serum, plasma, urine, and cerebrospinal fluid.

99. 99. The method of claim 98, wherein the mammalian subject is suffering from or at risk of developing an alternative pathway disease or disorder.

100. The method of claim 98 or 99, wherein the mammalian subject has been treated with a complement inhibitor, such as an alternative complement pathway inhibitor, such as a pro-factor D maturation inhibitor, such as a MASP-3 inhibitory antibody or its antigen-binding fragment.

101. The method of any one of claims 91 to 100, wherein the anti-human maturation factor D specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23.

102. The anti-human maturation factor D specific antibody or antigen-binding fragment thereof has six CDRs: (a) HC-CDR1 comprising the amino acid sequence XSXMGVS (SEQ ID NO: 65), in which X at position 1 is T, I, or S, and X at position 3 is G or I; (b) amino acid sequence (c) an amino acid sequence comprising: (wherein X at position 6 is R, G, or N, X at position 7 is S or Y, X at position 8 is F, I, or V, and X at position 10 is D or H); (d) an amino acid sequence (e) an LC-CDR2 comprising the amino acid sequence KVXNRFS (SEQ ID NO:69), (wherein X at position 3 is S or Y); and (f) an LC-CDR3 comprising the amino acid sequence FQGSHVPPT (SEQ ID NO:54). The method of any one of claims 91 to 101, comprising a binding domain comprising:

103. The anti-human maturation factor D specific antibody or antigen-binding fragment thereof has six CDRs: (a) HC-CDR1 comprising SEQ ID NO: 25, (b) HC-CDR2 comprising SEQ ID NO: 27, (c) HC-CDR3 comprising SEQ ID NO: 29, (d) LC-CDR1 comprising SEQ ID NO: 50, (e) LC-CDR2 comprising SEQ ID NO: 52, and (f) LC-CDR3 comprising SEQ ID NO:

54. The method of claim 102, comprising a binding domain comprising:

104. 1. A method for determining the presence or amount of pro-factor D in a test sample, comprising: (a) contacting a test sample with an anti-human profactor D specific monoclonal antibody or antigen-binding fragment thereof in an in vitro immunoassay; and (b) detecting the presence or amount of said antibody or antigen-binding fragment thereof bound to pro-factor D, wherein the presence of said binding indicates the presence or amount of pro-factor D in the sample. Including, The method, wherein the anti-human mature pro-factor D specific antibody or antigen-binding fragment thereof specifically binds to an epitope in the activation ("pro") peptide of human factor D designated as "APPRGR" (SEQ ID NO:4).

105. The method of claim 104, wherein the antibody or antigen-binding fragment thereof specifically binds to human profactor D (SEQ ID NO:2) and does not bind to human mature factor D (SEQ ID NO:3).

106. The method of claim 104 or 105, wherein the anti-human profactor D specific antibody or antigen-binding fragment thereof is immobilized on a substrate.

107. The method of any one of claims 104 to 106, wherein the immunoassay is an ELISA assay.

108. The method of any one of claims 104 to 107, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof is labeled with a detectable moiety, and step (b) comprises detecting the presence or amount of the detectable moiety.

109. The method of any one of claims 104 to 107, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof is naked (i.e., unlabeled) and the presence or amount of the antibody or fragment thereof bound to mature factor D is detected using a labeled antibody that binds to the anti-pro-factor D antibody.

110. The method of any one of claims 104 to 109, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof is immobilized (i.e., captured / coated) on a substrate and the bound pro-factor D is detected using a second antibody that binds to a different epitope of factor D.

111. 111. The method of any one of claims 104-110, wherein the test sample is a biological sample obtained from a mammalian subject, e.g., the biological sample is selected from the group consisting of blood, serum, plasma, urine, and cerebrospinal fluid.

112. The method of claim 111, wherein the mammalian subject is suffering from or at risk of developing an alternative pathway disease or disorder.

113. The method of claim 111 or 112, wherein the mammalian subject has been treated with a complement inhibitor, such as an alternative complement pathway inhibitor, such as a pro-factor D maturation inhibitor, such as a MASP-3 inhibitory antibody or its antigen-binding fragment.

114. The method of any one of claims 104 to 113, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 141, and LC-CDR-1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 147, wherein the CDRs are numbered according to the Kabat numbering system.

115. The method of any one of claims 104 to 113, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136 to 139, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 142 to 145, wherein the CDRs are numbered according to the Kabat numbering system.

116. The method of any one of claims 104 to 113, wherein the anti-human pro-factor D specific antibody or antigen-binding fragment thereof comprises a binding domain comprising HC-CDR-1, HC-CDR-2, and HC-CDR-3 in a heavy chain variable region selected from the group consisting of SEQ ID NO: 140 and SEQ ID NO: 141, and LC-CDR-1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NO: 146 and SEQ ID NO: 147, wherein the CDRs are numbered according to the Kabat numbering system.

117. The anti-human profactor D specific antibody or antigen-binding fragment thereof comprises the following six CDRs: (a) a CDR-H1 comprising SEQ ID NO: 167; (b) a CDR-H2 comprising SEQ ID NO: 169 or SEQ ID NO: 173; (c) a CDR-H3 comprising SEQ ID NO: 171 or SEQ ID NO: 174; (d) a CDR-L1 comprising SEQ ID NO: 194; (e) a CDR-L2 comprising SEQ ID NO: 196 or SEQ ID NO: 199; and (f) a CDR-L3 comprising SEQ ID NO: 198 or SEQ ID NO:

200. The method of any one of claims 104 to 113, comprising a binding domain comprising:

118. 1. A method for assessing the degree of alternative pathway complement (APC) activation in a test sample, comprising: (a) providing a test sample; (b) performing an immunoassay comprising at least one of the following: (i) capturing and detecting mature factor D in the test sample, wherein mature factor D is captured or detected with a mature factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but not pro-factor D; and / or (ii) capturing and detecting pro-factor D in the test sample, wherein pro-factor D is captured or detected with a pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but not mature factor D; and (c) comparing the level of mature factor D detected according to (b)(i) with a predetermined level or a control sample and / or comparing the level of pro-factor D detected according to (b)(ii) with a predetermined level or a control sample, wherein the detected level of mature factor D and / or pro-factor D in the test sample indicates the degree of alternative pathway complement activation. A method comprising:

119. The method of claim 118, wherein step (b)(i) comprises capturing mature factor D with a mature factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but not pro-factor D, and detecting with an antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D.

120. The method of claim 118, wherein step (b)(i) comprises capturing mature factor D with an anti-factor D antibody or an antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, and detecting with a mature factor D-specific monoclonal antibody or an antigen-binding fragment thereof that specifically binds to an epitope in "ILGGREA" (SEQ ID NO:5) present in mature factor D but not pro-factor D.

121. The method of any one of claims 118 to 120, wherein step (b)(ii) comprises capturing pro-factor D with a pro-factor D-specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but not mature factor D, and detecting with an antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D.

122. The method of any one of claims 118 to 120, wherein step (b)(ii) comprises capturing pro-factor D with an anti-factor D antibody or an antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, and detecting with a monoclonal antibody or an antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO:4) present in pro-factor D but not mature factor D.

123. The method of any one of claims 118 to 122, wherein the mature factor D specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in "ILGGREA" (SEQ ID NO: 5) present in mature factor D but does not bind to pro-factor D comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a heavy chain variable region selected from the group consisting of SEQ ID NOs: 12 to 17, and LC-CDR1, LC-CDR2, and LC-CDR3 in a light chain variable region selected from the group consisting of SEQ ID NOs: 18 to 23, wherein the CDRs are numbered according to the Kabat numbering system.

124. The method of any one of claims 118 to 123, wherein the pro-factor D specific monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the activation ("pro") peptide "APPRGR" (SEQ ID NO: 4) present in pro-factor D but not mature factor D, comprises a binding domain comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 136-141 and LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 142-147, wherein the CDRs are numbered according to the Kabat numbering system.

125. 125. The method of any one of claims 118 to 124, wherein said test sample is a biological sample obtained from a mammalian subject.

126. 126. The method of claim 125, wherein the biological sample comprises whole blood, serum, plasma, urine, or cerebrospinal fluid.

127. The method of any one of claims 118 to 126, wherein the test sample comprises a complement inhibitor, such as an alternative complement pathway inhibitor, such as a pro-factor D maturation inhibitor, such as a MASP-3 inhibitor.

128. The method of claim 125, wherein the mammalian subject has been treated with a complement inhibitor, such as an alternative complement pathway inhibitor, such as a pro-factor D maturation inhibitor, such as a MASP-3 inhibitor, and the degree of inhibition of the alternative pathway is measured using the assay.

129. 129. The method of claim 125 or 128, wherein the mammalian subject is a human subject.

130. The method of claim 129, wherein the human subject is suffering from, at risk of developing, or suspected of having an alternative pathway disease or disorder.

131. The alternative pathway disease or disorder is selected from the group consisting of paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple myelopathy, and glaucoma.

131. The method of claim 130, wherein the patient is selected from the group consisting of chronic sclerosis, Guillain-Barre syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft versus host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

132. The method of claim 128, wherein the control sample is a sample taken from the subject before treatment with a MASP-3 inhibitor or a sample taken at an earlier point during the course of treatment with a MASP-3 inhibitor.

133. The method of claim 128 or 132, wherein the MASP-3 inhibitor is a MASP-3 inhibitory antibody or its antigen-binding fragment.

134. The MASP-3 inhibitory antibody is a monoclonal antibody or an antigen-binding fragment thereof that binds to MASP-3; and (i) a binding domain comprising an HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 220, 222, 223, 225, 226, and 228, and an LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 221, 224, and 227, wherein the CDRs are numbered according to the Kabat numbering system; (ii) HC-CDR1 comprising SEQ ID NO: 229 (TDDIN); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 236 (LEDTY); and a HC-CDR3 comprising SEQ ID NO: 237 (LEDTY); a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 177 (SEQ ID NO: 179), an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 242 (KQSYNLYT); (iii) HC-CDR1 comprising SEQ ID NO: 230 (SYGMS); and a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 237 (GGEAMDY); and an HC-CDR3 comprising SEQ ID NO: 237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 241 (LVSKLDS), an LC-CDR2 comprising SEQ ID NO: 241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO: 243 (WQGTHFPWT); or (iv) HC-CDR1 comprising SEQ ID NO: 231 (GKWIE); or and a HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT).

134. The method of claim 133, comprising at least one of:

135. A method for monitoring the effectiveness of treatment with a MASP-3 inhibitory antibody or antigen-binding fragment thereof in a mammalian subject, comprising: (a) administering a dose of a MASP-3 inhibitory antibody or antigen-binding fragment thereof to a mammalian subject at a first time point; (b) assessing a first concentration of mature factor D and / or pro-factor D in a biological sample obtained from the subject after step (a); (c) treating the subject with the MASP-3 inhibitory antibody or antigen-binding fragment thereof at a second time point; (d) assessing a second concentration of mature factor D and / or pro-factor D in a biological sample obtained from the subject after step (c); and (e) comparing the level of mature factor D and / or pro-factor D assessed in step (b) with the level of mature factor D and / or pro-factor D assessed in step (d) to determine the efficacy of the MASP-3 inhibitory antibody or antigen-binding fragment thereof in the mammalian subject. A method comprising:

136. The method of claim 135, further comprising a step of adjusting the dose of the MASP-3 inhibitory antibody or its antigen-binding fragment.

137. The method of claim 136, wherein the dose of the MASP-3 inhibitory antibody or its antigen-binding fragment administered to the subject is increased if the level of the maturation factor D is higher than the control or reference standard.

138. The method of claim 136, wherein the dose of the MASP-3 inhibitory antibody or its antigen-binding fragment administered to the subject is increased if the level of pro-factor D is lower than the control or reference standard.

139. The method of claim 137 or 138, wherein when an increased dose of the MASP-3 inhibitory antibody or its antigen-binding fragment is administered to the subject, steps (b) to (e) are repeated to determine whether the increased dose is sufficient to adjust the levels of mature factor D and / or pro-factor D to the desired level compared to the respective control or reference standard.

140. The method of claim 135, wherein steps (b) and (d) comprise assessing the concentration of mature factor D in the biological sample by immunoassay.

141. The method of claim 140, wherein the immunoassay comprises (i) a first monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope comprising or consisting of amino acids ILGGREA (SEQ ID NO:5) in the N-terminal region of human maturation factor D and does not bind to human pro-factor D, and (ii) a second antibody or antigen-binding fragment thereof that binds to an epitope common to both human maturation factor D and human pro-factor D, wherein the first and second antibodies or antigen-binding fragments function together in the immunoassay to specifically detect or quantify the amount of mature factor D protein (SEQ ID NO:3) but not pro-factor D protein (SEQ ID NO:2) that may be present in the biological sample.

142. The method of claim 135, wherein steps (b) and (d) comprise assessing the concentration of pro-factor D in the biological sample by immunoassay.

143. The method of claim 142, wherein the immunoassay comprises (i) a first monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope comprising or consisting of amino acids APPRGR (SEQ ID NO:4) in the propeptide of human factor D and does not bind to human mature factor D, and (ii) a second antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the first and second antibodies or antigen-binding fragments function together in the immunoassay to specifically detect or quantify the amount of pro-factor D protein (SEQ ID NO:2) but not mature factor D protein (SEQ ID NO:3) that may be present in the biological sample.

144. The method of any one of claims 135 to 143, wherein said mammalian subject is a human subject.

145. The method of claim 144, wherein the human subject is suffering from or at risk of developing an alternative pathway disease or disorder.

146. The alternative pathway disease or disorder is selected from the group consisting of paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple myelopathy, and glaucoma.

146. The method of claim 145, wherein the patient is selected from the group consisting of chronic sclerosis, Guillain-Barré syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft versus host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

147. The method of claim 135, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding fragment.

148. The MASP-3 inhibitory antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof that binds to MASP-3; and (i) a binding domain comprising an HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 220, 222, 223, 225, 226, and 228, and an LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 221, 224, and 227, wherein the CDRs are numbered according to the Kabat numbering system; (ii) HC-CDR1 comprising SEQ ID NO: 229 (TDDIN); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 236 (LEDTY); and a HC-CDR3 comprising SEQ ID NO: 237 (LEDTY); a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 177 (SEQ ID NO: 179), an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 242 (KQSYNLYT); (iii) HC-CDR1 comprising SEQ ID NO: 230 (SYGMS); and a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 237 (GGEAMDY); and an HC-CDR3 comprising SEQ ID NO: 237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 241 (LVSKLDS), an LC-CDR2 comprising SEQ ID NO: 241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO: 243 (WQGTHFPWT); or (iv) HC-CDR1 comprising SEQ ID NO: 231 (GKWIE); or and a HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT).

148. The method of claim 147, comprising at least one of:

149. 1. A method of treating a mammalian subject suffering from or at risk of developing an alternative pathway disease or disorder, comprising administering to the subject: (i) the level of pro-factor D in one or more samples taken from the subject is low or decreased compared to a predetermined pro-factor D level or compared to the level of pro-factor D in one or more control samples; and / or (ii) the level of maturation factor D in one or more samples taken from the subject is elevated or increased compared to a predetermined level of maturation factor D or compared to the level of maturation factor D in one or more control samples. A method comprising the step of administering to the subject an MASP-3 inhibitory antibody or its antigen-binding fragment if the subject is determined to have MASP-3.

150. The method of claim 149, wherein the level of pro-factor D in one or more samples taken from the subject is determined by performing an immunoassay comprising the use of a pro-factor D-specific monoclonal antibody.

151. The method of claim 150, wherein the immunoassay comprises (i) a first monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope comprising or consisting of amino acids APPRGR (SEQ ID NO:4) in the propeptide of human factor D and does not bind to human mature factor D, and (ii) a second antibody or antigen-binding fragment thereof that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the first and second antibodies or antigen-binding fragments function together in the immunoassay to specifically detect or quantify the amount of pro-factor D protein (SEQ ID NO:2) but not mature factor D protein (SEQ ID NO:3) that may be present in the sample.

152. The method of claim 149, wherein the level of mature factor D in one or more samples taken from the subject is determined by performing an immunoassay comprising the use of a mature factor D-specific monoclonal antibody or an antigen-binding fragment thereof.

153. The method of claim 152, wherein the immunoassay comprises (i) a first monoclonal antibody, or antigen-binding fragment thereof, that specifically binds to an epitope comprising or consisting of amino acids ILGGREA (SEQ ID NO:5) in the N-terminal region of human mature factor D and does not bind to human pro-factor D, and (ii) a second antibody, or antigen-binding fragment thereof, that binds to an epitope common to both human mature factor D and human pro-factor D, wherein the first and second antibodies, or antigen-binding fragments thereof, function together in the immunoassay to specifically detect or quantify the amount of mature factor D protein (SEQ ID NO:3) but not pro-factor D protein (SEQ ID NO:2) that may be present in the sample.

154. The method of any one of claims 149-153, wherein said mammalian subject is a human subject.

155. The human subject is furthermore afflicted by a disease, such as paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (AMD, including wet AMD and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uraemic syndrome (HUS), atypical hemolytic uraemic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple sclerosis, Guillain-Barré syndrome, Alzheimer's disease, or the like. The method of claim 154, wherein the patient is suffering from or at risk of developing an alternative pathway disease or disorder selected from the group consisting of chronic myocardial infarction, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft versus host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

156. The method of claim 149, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding fragment.

157. The MASP-3 inhibitory antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof that binds to MASP-3; and (i) a binding domain comprising an HC-CDR1, HC-CDR2, and HC-CDR3 of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 220, 222, 223, 225, 226, and 228, and an LC-CDR1, LC-CDR2, and LC-CDR3 of a light chain variable region selected from the group consisting of SEQ ID NOs: 221, 224, and 227, wherein the CDRs are numbered according to the Kabat numbering system; (ii) HC-CDR1 comprising SEQ ID NO: 229 (TDDIN); a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 236 (LEDTY); and a HC-CDR3 comprising SEQ ID NO: 237 (LEDTY); a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 177 (SEQ ID NO: 179), an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 242 (KQSYNLYT); (iii) HC-CDR1 comprising SEQ ID NO: 230 (SYGMS); and a heavy chain variable region comprising an HC-CDR2 comprising SEQ ID NO: 237 (GGEAMDY); and an HC-CDR3 comprising SEQ ID NO: 237 (GGEAMDY); a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 241 (LVSKLDS), an LC-CDR2 comprising SEQ ID NO: 241 (LVSKLDS), and an LC-CDR3 comprising SEQ ID NO: 243 (WQGTHFPWT); or (iv) HC-CDR1 comprising SEQ ID NO: 231 (GKWIE); or and a HC-CDR3 comprising SEQ ID NO: 238 (SEDV); and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT).

157. The method of claim 156, comprising at least one of the following:

158. A pharmaceutical composition comprising a MASP-3 inhibitory antibody or its antigen-binding fragment in an aqueous solution comprising a buffer system having a pH of 6.0±5%, 20±5% mM histidine, 100±5% mg / mL sucrose, and 0.035%±5% polysorbate 80, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is contained at a concentration of 110 mg / mL±5%, and the MASP-3 inhibitory antibody or its antigen-binding fragment has an HC-CDR1 comprising SEQ ID NO: 231 (GKWIE), or and a light chain variable region comprising an LC-CDR1 comprising SEQ ID NO: 239, an LC-CDR2 comprising SEQ ID NO: 178 (WASTRES), and an LC-CDR3 comprising SEQ ID NO: 244 (KQSYNIPT).

159. The pharmaceutical composition of claim 158, which is sterile.

160. The pharmaceutical composition of claim 158, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a heavy chain variable region that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to SEQ ID NO: 226 or SEQ ID NO: 227, and a light chain variable region that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to SEQ ID NO:

227.

161. A pharmaceutical composition described in any one of claims 158 to 160, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is selected from the group consisting of a human antibody, a humanized antibody, a chimeric antibody, a murine antibody, and an antigen-binding fragment of any of them.

162. A pharmaceutical composition described in any one of claims 158 to 161, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment is selected from the group consisting of a single-chain antibody, an ScFv, a Fab fragment, a Fab' fragment, an F(ab')2 fragment, a monovalent antibody lacking the hinge region, and a full-length antibody.

163. A pharmaceutical composition described in any one of claims 158 to 162, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment further comprises an immunoglobulin constant region.

164. A pharmaceutical composition described in any one of claims 158 to 163, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a human IgG4 constant region.

165. The pharmaceutical composition of claim 164, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a human IgG4 constant region having an S228P mutation.

166. The pharmaceutical composition of claim 164 or 165, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment contains a mutation that promotes FcRn interaction at low pH.

167. The pharmaceutical composition of claim 166, wherein the MASP-3 inhibitory antibody or its antigen-binding fragment comprises a human IgG4 constant region as shown in SEQ ID NO:

245.

168. 168. An article of manufacture containing the pharmaceutical composition of any one of claims 158-167.

169. The article of manufacture described in claim 168, wherein the MASP-3 inhibitory antibody or antigen-binding fragment thereof is in a unit dosage form of 10 mg to 1000 mg suitable for therapeutic administration to a human subject.

170. The article of manufacture of claim 169, wherein the MASP-3 inhibitory antibody or antigen-binding fragment thereof is in a unit dosage form of 100 mg to 200 mg suitable for therapeutic administration to a human subject.

171. The article of manufacture of any one of claims 168-170, comprising a container and a label or package insert on or associated with the container.

172. 172. The article of manufacture of claim 171, wherein the container is selected from the group consisting of a bottle, an ampoule, a pouch (e.g., an intravenous infusion bag), a vial, a syringe, and a cartridge.

173. A pharmaceutical composition of any one of claims 158-167 or an article of manufacture of any one of claims 168-172 for use in treating a subject suffering from or at risk of developing an alternative pathway disease or disorder.

174. The alternative pathway disease or disorder is selected from the group consisting of paroxysmal nocturnal hemoglobinuria (PNH), age-related macular degeneration (including AMD, wet AMD and dry AMD), ischemia-reperfusion injury, arthritis, disseminated intravascular coagulation, thrombotic microangiopathy (including hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), or transplant-associated TMA), asthma, dense deposit disease, microimmune necrotizing crescentic glomerulonephritis, traumatic brain injury, aspiration pneumonia, endophthalmitis, neuromyelitis optica, Behcet's disease, multiple sclerosis, 174. The pharmaceutical composition or article of manufacture of claim 173, wherein the disease is selected from the group consisting of Guillain-Barre syndrome, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), lupus nephritis, systemic lupus erythematosus (SLE), diabetic retinopathy, uveitis, chronic obstructive pulmonary disease (COPD), C3 glomerulopathy, transplant rejection, graft versus host disease (GVHD), hemodialysis, sepsis, systemic inflammatory response syndrome (SIRS), acute respiratory distress syndrome (ARDS), ANCA vasculitis, antiphospholipid syndrome, atherosclerosis, IgA nephropathy, and myasthenia gravis.

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