Anti-human factor d antibody constructs and uses thereof

Abstract

The present application relates to anti-human factor D (FD) antibody constructs. The present application also provides methods of using the anti-human FD antibody constructs to treat complement-mediated diseases, pharmaceutical compositions of the anti-human FD antibody constructs, and methods of making the anti-human FD antibody constructs.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to International Patent Application No. PCT / CN2023 / 116638 and International Patent Application No. PCT / CN2023 / 116639, filed on September 3, 2023, the contents of which are incorporated herein by reference in their entirety.

[0003] Reference to the electronic sequence list

[0004] The contents of the electronic sequence list (792252001141seqlist.xml; size: 370,943 bytes; and creation date: August 9, 2024) are incorporated herein by reference in their entirety. Technical Field

[0005] This invention relates to anti-human factor D (FD) antibody constructs and their uses. Background Technology

[0006] The complement system is part of the innate immune system and plays a crucial role in host defense. Complement also plays a pathogenic role in human inflammatory diseases. Activation of the complement system occurs through three distinct pathways: the classical pathway (CP), the lectin pathway (LP), and the alternative pathway (AP). CP is triggered by antigen-antibody binding. LP is triggered when mannose-binding lectin (MBL) interacts with sugar molecules on the surface of microorganisms. Activation of either pathway leads to the assembly of CP C3 convertase C4b2a, but direct cleavage of C3 by MBL-associated serine proteases can also occur. AP is a self-amplifying loop driven by AP C3 convertase C3bBb. AP activation can be secondary to CP or LP activation, or it can be initiated independently. In the latter case, low levels of spontaneous C3 “tick-over” produce initial C3bBb, which, in the absence of proper regulation, rapidly triggers AP. Therefore, it is generally accepted that AP activation on non-self surfaces with no or insufficient negative regulation is considered a default process, and autologous cells typically avoid this outcome by utilizing various membrane-bound and liquid-phase complement repressor proteins. Under certain conditions, altered, damaged, or stressed autologous cells and tissues can also activate AP and cause inflammatory damage.

[0007] Factor D (FD) is an essential enzyme for complement activation in the complement pathway (AP). After factor B binds to C3b, FD cleaves factor B, producing the active C3 convertase C3bBb. Factor D is a serine protease of approximately 24 kDa, produced from a factor D precursor via the enzymatic action of mannose-binding lectin-associated serine protease-3 (MASP-3), and circulates in the blood as a constitutively active enzyme after production. The concentration of factor D in the blood is relatively low (approximately 2 μg / ml) compared to other complement proteins. While this latter fact suggests that therapeutic inhibition of factor D activity in the blood is possible and likely easily achievable, previous studies have shown that factor D has a rapid turnover rate; therefore, the consensus in complement research is that systemic blockade of factor D is not feasible.

[0008] Small molecule antagonists of FD have been tested in clinical trials and have shown good AP inhibition and efficacy. However, this treatment regimen requires high doses and frequent dosing, and therefore hepatotoxicity has been reported. Anti-FD biologics for the treatment of macular disease have been evaluated in clinical trials. Topical application of the anti-FD Fab fragment to the vitreous humor is used to maintain good pharmacokinetics. Clinical trials showed good efficacy in Phase II trials, but failed to meet the primary endpoint in Phase III trials.

[0009] All references cited in this article, including patent applications, patent publications, and Genbank accession numbers, are incorporated in full as if each individual reference were specifically and individually cited. Summary of the Invention

[0010] This application provides anti-human FD antibody constructs, including mature anti-human FD antibody constructs with pH-dependent factor D binding and / or affinity.

[0011] In some aspects, this document provides an isolated antibody construct (anti-human FD antibody construct) comprising an antibody moiety specifically binding to human factor D (anti-human FD antibody moiety), wherein the anti-human FD antibody moiety comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), wherein: (i) the VH comprises heavy chain CDR1 (“H-CDR1”), the H-CDR1 comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof comprising up to 3 amino acid variations; heavy chain CDR2 (“H-CDR2”), the H-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 or a variant thereof comprising up to 3 amino acid variations; and heavy chain CDR3 (“H-CDR3”), the H-CDR3 comprising the amino acid sequence of SEQ ID NO: 3 or a variant thereof comprising up to 3 amino acid variations; and (ii) the VL comprises light chain CDR1 (“L-CDR1”), the L-CDR1 comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof comprising up to 3 amino acid variations; and (ii) the VL comprises light chain CDR1 (“L-CDR1”), the L-CDR1 comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof comprising up to 3 amino acid variations; and (iii) the VL comprises light chain CDR1 (“L-CDR1”), the L-CDR1 comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof comprising up to 3 amino acid variations; and (iv ... The amino acid sequence of SEQ ID NO: 4 or a variant thereof containing up to 3 amino acid variations; light chain CDR2 (“L-CDR2”), said L-CDR2 containing the amino acid sequence of SEQ ID NO: 5 or a variant thereof containing up to 3 amino acid variations; and light chain CDR3 (“L-CDR3”), said L-CDR3 containing the amino acid sequence of SEQ ID NO: 6 or a variant thereof containing up to 3 amino acid variations. In some embodiments, (i) the VH comprises: H-CDR1 containing the amino acid sequence DTYVH (SEQ ID NO: 1); H-CDR2 containing the amino acid sequence RIDP-X1-X2-G-X3-T-X4-F-X5-PRFQA (SEQ ID NO: 9), wherein X1 is A or H, X2 is N, S or Y, X3 is L or H, X4 is T or H, and X5 is D, V, L or H; and H-CDR3 containing the amino acid sequence AMEY (SEQ ID NO: 3); and (ii) the VL comprises: L-CDR1 containing the amino acid sequence SA-X6-SDVS-X7-MY (SEQ ID NO: 10), wherein X6 is R, N or S, and X7 is Y, D or V; and H-CDR2 containing the amino acid sequence X8-TSNLAS (SEQ ID NO: 10). L-CDR2 (252), wherein X8 is D or H; and L-CDR3 containing the amino acid sequence QQWSS-X9-PPW-X10 (SEQ ID NO: 11), wherein X9 is Y or H and X10 is T, R or H.

[0012] In some embodiments of the anti-human FD antibody construct isolated according to any of the above, (1) the VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 1, H-CDR2 containing the amino acid sequence of SEQ ID NO: 2, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 4, L-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 6; (2) the VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 20, H-CDR2 containing the amino acid sequence of SEQ ID NO: 21, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 22; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 23, L-CDR2 containing the amino acid sequence of SEQ ID NO: 24, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 25. (2) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 36, H-CDR2 containing the amino acid sequence of SEQ ID NO: 37, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 38; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 39, L-CDR2 containing the amino acid sequence of SEQ ID NO: 40, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 41; (4) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 44, H-CDR2 containing the amino acid sequence of SEQ ID NO: 45, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 46; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 47, L-CDR2 containing the amino acid sequence of SEQ ID NO: 48, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 49. L-CDR3 of the amino acid sequence of SEQ ID NO: 59; (5) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 52, H-CDR2 containing the amino acid sequence of SEQ ID NO: 53, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 54; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57;(6) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 63, L-CDR2 containing the amino acid sequence of SEQ ID NO: 64, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 65; (7) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 71, L-CDR2 containing the amino acid sequence of SEQ ID NO: 72, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 63. (8) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 76, H-CDR2 containing the amino acid sequence of SEQ ID NO: 77, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 78; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 79, L-CDR2 containing the amino acid sequence of SEQ ID NO: 80, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 81; (9) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 84, H-CDR2 containing the amino acid sequence of SEQ ID NO: 85, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 86; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89. L-CDR3 of amino acid sequence 89; (10) The VH includes H-CDR1 containing amino acid sequence of SEQ ID NO: 92, H-CDR2 containing amino acid sequence of SEQ ID NO: 93, and H-CDR3 containing amino acid sequence of SEQ ID NO: 94; and the VL includes L-CDR1 containing amino acid sequence of SEQ ID NO: 95, L-CDR2 containing amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing amino acid sequence of SEQ ID NO: 97;(11) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 100, H-CDR2 containing the amino acid sequence of SEQ ID NO: 101, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 102; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (12) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 108, H-CDR2 containing the amino acid sequence of SEQ ID NO: 109, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 110; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 105. (13) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 116, H-CDR2 containing the amino acid sequence of SEQ ID NO: 117, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 118; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (14) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, H-CDR2 containing the amino acid sequence of SEQ ID NO: 125, H-CDR3 containing the amino acid sequence of SEQ ID NO: 126 ... L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126; (15) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126;(16) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 4, L-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 6; (17) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 1, H-CDR2 containing the amino acid sequence of SEQ ID NO: 2, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 129. (18) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89; (19) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, H-CDR2 containing the amino acid sequence of SEQ ID NO: 128, H-CDR3 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, H-CDR3 containing the amino acid sequence of SEQ ID NO: 129, H-CDR3 containing the amino acid sequence of SEQ ID NO: 129, H-CDR2 ... L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (20) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97;(21) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113; (22) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 128. (23) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57; (24) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 87. L-CDR3 of the amino acid sequence NO: 89; (25) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105;(26) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97; (27) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 111. L-CDR3 of the amino acid sequence SEQ ID NO: 60; (28) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (29) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 70. L-CDR3 of the amino acid sequence of SEQ ID NO: 57; (30) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89;(31) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (32) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 95. (33) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113; (34) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, H-CDR2 containing the amino acid sequence of SEQ ID NO: 113, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 113; L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (35) the VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 206, H-CDR2 containing the amino acid sequence of SEQ ID NO: 207, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 208; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 209, L-CDR2 containing the amino acid sequence of SEQ ID NO: 210, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 211;(36) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 234, H-CDR2 containing the amino acid sequence of SEQ ID NO: 235, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 236; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 237, L-CDR2 containing the amino acid sequence of SEQ ID NO: 238, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 239; (37) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 240, H-CDR2 containing the amino acid sequence of SEQ ID NO: 241, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 242; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 243, L-CDR2 containing the amino acid sequence of SEQ ID NO: 244, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 244. L-CDR3 of the amino acid sequence 245; or (38) the VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 246, H-CDR2 containing the amino acid sequence of SEQ ID NO: 247, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 248; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 249, L-CDR2 containing the amino acid sequence of SEQ ID NO: 250, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 251.

[0013] In some embodiments of the isolated anti-human FD antibody construct according to any of the above, the anti-human FD antibody portion comprises: VH, the VH comprising the amino acid sequence of SEQ ID NO: 7, or a variant thereof having at least about 80% amino acid sequence homology with SEQ ID NO: 7; and VL, the VL comprising the amino acid sequence of SEQ ID NO: 8, or a variant thereof having at least about 80% amino acid sequence homology with SEQ ID NO: 8. In some embodiments, i) the amino acid residue at position 24 of VH is A or T; ii) the amino acid residue at position 54 of VH is A or H; iii) the amino acid residue at position 55 of VH is N, S, or Y; iv) the amino acid residue at position 57 of VH is H or L; v) the amino acid residue at position 61 of VH is D, H, L, or V; vi) the amino acid residue at position 74 of VH is K or T; vii) the amino acid residue at position 26 of VL is N, R, or S; viiii) the amino acid residue at position 31 of VL is D, V, or Y; ix) the amino acid residue at position 49 of VL is D or H; x) the amino acid residue at position 70 of VL is F or Y; xi) the amino acid residue at position 93 of VL is H or Y; and / or xii) the amino acid residue at position 97 of VL is T, H, or R; wherein the numbering is based on the Kabat numbering system.

[0014] In some embodiments of the anti-human FD antibody construct isolated according to any of the above, (1) the VH comprises the amino acid sequence of SEQ ID NO: 7, and the VL comprises the amino acid sequence of SEQ ID NO: 8; (2) the VH comprises the amino acid sequence of SEQ ID NO: 18, and the VL comprises the amino acid sequence of SEQ ID NO: 19; (3) the VH comprises the amino acid sequence of SEQ ID NO: 26, and the VL comprises the amino acid sequence of SEQ ID NO: 27; (4) the VH comprises the amino acid sequence of SEQ ID NO: 34, and the VL comprises the amino acid sequence of SEQ ID NO: 35; (5) the VH comprises the amino acid sequence of SEQ ID NO: 42, and the VL comprises the amino acid sequence of SEQ ID NO: 43; (6) the VH comprises the amino acid sequence of SEQ ID NO: 50, and the VL comprises the amino acid sequence of SEQ ID NO: 51; (7) the VH comprises the amino acid sequence of SEQ ID NO: 58, and the VL comprises the amino acid sequence of SEQ ID NO: 59. (8) The VH contains the amino acid sequence of SEQ ID NO: 66, and the VL contains the amino acid sequence of SEQ ID NO: 67; (9) The VH contains the amino acid sequence of SEQ ID NO: 74, and the VL contains the amino acid sequence of SEQ ID NO: 75; (10) The VH contains the amino acid sequence of SEQ ID NO: 82, and the VL contains the amino acid sequence of SEQ ID NO: 83; (11) The VH contains the amino acid sequence of SEQ ID NO: 90, and the VL contains the amino acid sequence of SEQ ID NO: 91; (12) The VH contains the amino acid sequence of SEQ ID NO: 98, and the VL contains the amino acid sequence of SEQ ID NO: 99; (13) The VH contains the amino acid sequence of SEQ ID NO: 106, and the VL contains the amino acid sequence of SEQ ID NO: 107; (14) The VH contains the amino acid sequence of SEQ ID NO: 114, and the VL contains the amino acid sequence of SEQ ID NO: 107. (15) The VH contains the amino acid sequence of SEQ ID NO: 122, and the VL contains the amino acid sequence of SEQ ID NO: 123; (16) The VH contains the amino acid sequence of SEQ ID NO: 156, and the VL contains the amino acid sequence of SEQ ID NO: 157;(17) The VH contains the amino acid sequence of SEQ ID NO: 158, and the VL contains the amino acid sequence of SEQ ID NO: 159; (18) The VH contains the amino acid sequence of SEQ ID NO: 160, and the VL contains the amino acid sequence of SEQ ID NO: 161; (19) The VH contains the amino acid sequence of SEQ ID NO: 162, and the VL contains the amino acid sequence of SEQ ID NO: 163; (20) The VH contains the amino acid sequence of SEQ ID NO: 164, and the VL contains the amino acid sequence of SEQ ID NO: 165; (21) The VH contains the amino acid sequence of SEQ ID NO: 166, and the VL contains the amino acid sequence of SEQ ID NO: 167; (22) The VH contains the amino acid sequence of SEQ ID NO: 168, and the VL contains the amino acid sequence of SEQ ID NO: 169; (23) The VH contains the amino acid sequence of SEQ ID NO: 169; (24) The VH contains the amino acid sequence of SEQ ID NO: 172, and the VL contains the amino acid sequence of SEQ ID NO: 173; (25) The VH contains the amino acid sequence of SEQ ID NO: 176, and the VL contains the amino acid sequence of SEQ ID NO: 177; (26) The VH contains the amino acid sequence of SEQ ID NO: 178, and the VL contains the amino acid sequence of SEQ ID NO: 179; (27) The VH contains the amino acid sequence of SEQ ID NO: 180, and the VL contains the amino acid sequence of SEQ ID NO: 181; (28) The VH contains the amino acid sequence of SEQ ID NO: 182, and the VL contains the amino acid sequence of SEQ ID NO: 183; (29) The VH contains the amino acid sequence of SEQ ID NO: 184, and the VL contains the amino acid sequence of SEQ ID NO: 175; (30) The VH contains the amino acid sequence of SEQ ID NO: 186, and the VL contains the amino acid sequence of SEQ ID NO: 187; (31) The VH contains the amino acid sequence of SEQ ID NO: 190, and the VL contains the amino acid sequence of SEQ ID NO: 191; (32) The VH contains the amino acid sequence of SEQ ID NO: 192, and the VL contains the amino acid sequence of SEQ ID NO: 193;(33) The VH contains the amino acid sequence of SEQ ID NO: 194, and the VL contains the amino acid sequence of SEQ ID NO: 195; (34) The VH contains the amino acid sequence of SEQ ID NO: 196, and the VL contains the amino acid sequence of SEQ ID NO: 197; (35) The VH contains the amino acid sequence of SEQ ID NO: 198, and the VL contains the amino acid sequence of SEQ ID NO: 199; (36) The VH contains the amino acid sequence of SEQ ID NO: 212, and the VL contains the amino acid sequence of SEQ ID NO: 213; (37) The VH contains the amino acid sequence of SEQ ID NO: 253, and the VL contains the amino acid sequence of SEQ ID NO: 254; (38) The VH contains the amino acid sequence of SEQ ID NO: 255, and the VL contains the amino acid sequence of SEQ ID NO: 256; (39) The VH contains the amino acid sequence of SEQ ID NO: 197. (40) The VH contains the amino acid sequence of SEQ ID NO: 257, and the VL contains the amino acid sequence of SEQ ID NO: 289; or (41) The VH contains the amino acid sequence of SEQ ID NO: 216, and the VL contains the amino acid sequence of SEQ ID NO: 214.

[0015] In some embodiments of any of the isolated anti-human FD antibody constructs described above, the anti-human FD antibody portion cross-reacts with cynomolgus factor D (cyno FD).

[0016] In some embodiments of any of the isolated anti-human FD antibody constructs described above, the isolated anti-human FD antibody moiety is pH-dependent, wherein the anti-human FD antibody moiety binds more strongly to human FD at neutral pH than at acidic pH.

[0017] In some embodiments of the isolated anti-human FD antibody construct according to the above description, the isolated anti-human FD antibody moiety is selected from the group consisting of: full-length antibody, Fab, Fab', F(ab)2, F(ab')2, scFv, and combinations thereof. In some embodiments, the anti-human FD antibody moiety is a full-length antibody (anti-human FD full-length antibody). In some embodiments, the anti-human FD full-length antibody comprises a heavy chain constant region derived from IgG4 (e.g., human IgG4). In some embodiments, the heavy chain constant region comprises the amino acid sequence of any one of SEQ ID NO: 132-139. In some embodiments, the anti-human FD antibody moiety is scFv (anti-human FD scFv). In some embodiments, the anti-human FD scFv comprises the amino acid sequence of SEQ ID NO: 219 or 220.

[0018] In some embodiments of the isolated anti-human FD antibody construct according to any of the above-described embodiments, the isolated anti-human FD antibody construct further comprises a second antibody portion that specifically recognizes a component of the complement pathway. In some embodiments, the second antibody portion is selected from the group consisting of: full-length antibody, Fab, Fab', F(ab)2, F(ab')2, scFv, sdAb, and combinations thereof. In some embodiments, the component of the complement pathway is complement component 2 (C2). In some embodiments, the component of the complement pathway is complement component 5 (C5). In some embodiments, the anti-human FD antibody portion is scFv (scFv1), and the second antibody portion is scFv (scFv2). In some embodiments, the isolated anti-human FD antibody construct comprises, from the N-terminus to the C-terminus: (i) scFv1-optional linker-scFv2; (ii) scFv2-optional linker-scFv1; (iii) scFv1-optional linker-Fc domain-optional linker-scFv2; or (iv) scFv2-optional linker-Fc domain-optional linker-scFv1. In some embodiments, i) the linker comprises the amino acid sequence of any one of SEQ ID NO: 221-229; and / or ii) the Fc domain comprises the amino acid sequence of SEQ ID NO: 230.

[0019] Also provided are isolated nucleic acids encoding any of the isolated anti-human FD antibody constructs described above. Additionally, a vector comprising any of the isolated nucleic acids described herein is provided. In some embodiments, the vector is a viral vector, such as an adeno-associated virus (AAV) vector or a lentiviral vector. A host cell is also provided comprising any of the isolated nucleic acids described herein, or any of the vectors described herein.

[0020] On the other hand, a method is provided for preparing an isolated anti-human FD antibody construct (e.g., any of the anti-human FD antibody constructs described herein), the method comprising: i) culturing a host cell containing any of the isolated nucleic acids or any of the vectors described herein, or any of the host cells described herein, under conditions suitable for expressing the isolated anti-human FD antibody construct; and ii) obtaining the expressed isolated anti-human FD antibody construct from the host cell.

[0021] On the other hand, a pharmaceutical composition is provided comprising any of the isolated anti-human FD antibody constructs described above, any of the isolated nucleic acids described above, or any of the vectors described above, and a pharmaceutically acceptable carrier.

[0022] On the other hand, a method for treating a complement-mediated disease in an individual is provided, the method comprising administering to the individual an effective amount of any of the pharmaceutical compositions described herein. In some embodiments, the complement-mediated disease is selected from the group consisting of: macular degeneration (MD), age-related macular degeneration (AMD), ischemia-reperfusion injury, arthritis, rheumatoid arthritis, lupus, ulcerative colitis, stroke, postoperative systemic inflammatory syndrome, asthma, allergic asthma, chronic obstructive pulmonary disease (COPD), paroxysmal nocturnal hemoglobinuria (PNH) syndrome, autoimmune hemolytic anemia (AIHA), and Gaucher disease. Diseases, myasthenia gravis, neuromyelitis optica (NMO), multiple sclerosis, delayed graft function, antibody-mediated rejection, atypical hemolytic uremic syndrome (aHUS), central retinal vein occlusion (CRVO), central retinal artery occlusion (CRAO), epidermolysis bullosa, sepsis, septic shock, organ transplantation, inflammation, C3 glomerulonephritis (C3G), membranous nephropathy, IgA nephropathy (IgAN), glomerulonephritis, thrombotic microangiopathy secondary to systemic lupus erythematosus (SLE-TMA), antineutrophil cytoplasmic antibody (ANCA)-mediated vasculitis, Shiga toxin-induced HUS, antiphospholipid antibody-induced abortion, graft-versus-host disease (GvHD), bullous pemphigoid, hidradenitis suppurativa, herpetic dermatitis, Sweet's syndrome Syndrome), gangrenous pyoderma, palmoplantar pustulosis and pustular psoriasis, rheumatoid neutrophilic dermatitis, subcorneal pustular dermatitis, intestinal-associated dermatitis-arthritis syndrome, neutrophilic eccrine sweat gland inflammation, linear IgA disease, and any combination thereof.

[0023] On the other hand, a method for reducing the complement system activity of an individual is provided, the method comprising administering an effective amount of any of the above-described pharmaceutical compositions to the individual.

[0024] These and other aspects and advantages of the invention will become apparent from the following detailed description and the appended claims. It should be understood that one, some, or all of the features of the various embodiments described herein can be combined to form other embodiments of the invention. Attached Figure Description

[0025] Figure 1 The binding activity of representative anti-FD scFv variants to human FD was depicted. z16-p2 is a parental, humanized anti-FD scFv. The z16-p2 clone was used as a reference anti-human FD control. Unrelated antibody clones were used as negative controls.

[0026] Figure 2A and 2B The binding activity of a representative anti-FD scFv variant to cynomolgus monkey FD was depicted. z16-p2 is a parental, humanized anti-FD scFv. The z16-p2 clone was used as a reference anti-human FD control. Unrelated antibody clones were used as negative controls.

[0027] Figure 3A The binding activity of representative anti-FD IgG4 monoclonal antibody (mAb) variants to human FD was described. Figure 3B The binding activity of a representative IgG4 mAb variant with cynomolgus monkey FD was depicted. The parental z16-p2 IgG4 form was used as a control.

[0028] Figure 4 Biolayer interference (BLI) sensing maps depicting the pH binding of engineered anti-FD variants to human and cynomolgus monkey FD.

[0029] Figure 5 The inhibitory effect of the affinity-matured anti-FD mAb variant on 50% NHS (normal human serum)-induced rabbit RBC lysis was depicted. The parental humanized z16-p2 clone was used as a control. Distilled water was used as a positive control. EDTA was used as a negative control.

[0030] Figure 6 The inhibitory effect of affinity-matured FD-resistant mAb variants on LPS-induced C3b deposition (in 10% NHS) was depicted. A parental z16-p2 clone was used as a control.

[0031] Figure 7 The inhibitory effect of pH-bound engineered anti-FD mAb on 30% NHS-induced rabbit RBC lysis was depicted. Parental affinity-matured mAb-42 was used as a control.

[0032] Figure 8 The inhibitory effect of pH-bound engineered anti-FD mAb on LPS-induced C3b deposition (in 10% NHS) was depicted. The parental affinity-matured mAb-42 was used as a control.

[0033] Figure 9 The FcRn binding optimization mutations used to enhance IgG recycling and reduce serum antigen levels were described.

[0034] Figure 10 The method of using BLI to optimize FcRn binding for antigen clearance was described. All tests were performed in duplicate.

[0035] Figure 11A A graph depicts the change in total plasma human IgG4 levels over time in hFD Scid / FcRn mice injected with 40 mg / kg mAb42-IgG4P (WT) or mAb42pH FcRn binding variants (Mut, YEY, YPY, N3E, and YTE). Figure 11B A graph depicts the change in total plasma hFD levels over time in hFD Scid / FcRn mice injected with 40 mg / kg mAb42-IgG4P (WT) or mAb42pH FcRn binding variants (Mut, YEY, YPY, N3E, and YTE). Figure 11C The total plasma IgG4 mAb levels over time were plotted in hFD Scid / FcRn mice injected with 40 mg / kg mAb42-IgG4P (WT) or mAb42pH FcRn binding variants (Mut, YEY, YPY, N3E, and YTE). Each test was performed in duplicate.

[0036] Figure 12A A graph depicts the total plasma IgG4 levels of mAb42-IgG4P-LA (42WT) and mAb42pH-IgG4P-N3E (N3E) in hFD Scid / FcRn mice injected with 40 mg / kg of any mAb. Figure 12B A graph depicting the total free plasma IgG4 levels at 42WT and N3E in hFD Scid / FcRn mice injected with 40 mg / kg of any mAb is presented. Figure 12C A graph depicts the change in total hFD levels over time in hFD Scid / FcRn mice injected with 40 mg / kg 42WT or N3E mAb. Figure 12D The ratio of total plasma IgG4 mAb to total plasma hFD in hFD Scid / FcRn mice injected with 40 mg / kg 42WT or N3E mAb was plotted over time. All tests were performed in duplicate. Detailed Implementation

[0037] This invention provides various anti-human FD antibody constructs, wherein the anti-human FD antibody constructs comprise an antibody portion (anti-human FD antibody portion) that specifically binds to human factor D. The anti-human FD antibody constructs described herein possess one or more of the following desirable properties: i) reduced immunogenicity (e.g., compared to parental mouse Ab); ii) reduced N-linked glycosylation potential; iii) modified cross-reactivity with cynomolgus monkey FD (e.g., affinity-matured variants), enabling extrapolation of monkey studies to humans; iv) maintaining or modifying binding affinity to human FD (e.g., affinity-matured variants, humanized variants, or pH-dependent FD-binding variants); v) some variants exhibit strong binding to FD at neutral pH (e.g., approximately pH 7.4; e.g., in plasma) but weaker binding to FD at acidic pH (e.g., approximately pH 5.8; e.g., in endosomes), allowing the anti-FD antibody to dissociate from FD in the acidic environment of endosomes and enabling the free anti-FD antibody to recirculate back to the extracellular space to bind to other free FD in plasma; vi) some variants exhibit strong binding affinity at pH 5.8 and / or pH 5.4. 7.4 Optimized FcRn binding, thereby improving antibody recycling, prolonging serum half-life (high FcRn binding at pH 5.8), and / or improving antigen clearance activity (to reduce free FD; high FcRn binding at pH 7.4); vii) exhibited strong inhibition of normal human serum (NHS)-induced rabbit RBC lysis and LPS-induced C3b deposition, reflecting the antibody's potent efficacy in inhibiting alternative pathways; and viii) demonstrated excellent in vivo pharmacokinetics (longer and / or higher antibody levels) and antigen clearance activity (lower FD accumulation in plasma).

[0038] I. Definition

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar to or equivalent to those described and used herein may be used in the practice or testing of this invention, exemplary methods and materials are described.

[0040] The term "antibody" is used in its broadest sense herein and encompasses a wide range of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, and their antigen-binding fragments, as long as they exhibit the desired antigen-binding activity. The term "antibody portion" refers to a full-length antibody or its antigen-binding fragment.

[0041] An "antibody" can refer to an immunoglobulin molecule or fragment thereof (including the basic 4-chain antibody unit) that can specifically bind to a specific epitope of an antigen. Antibodies can be complete immunoglobulins derived from natural or recombinant sources, and can be the immunoreactive portion of a complete immunoglobulin. The antibodies used in this invention can exist in various forms, including, for example, polyclonal antibodies, monoclonal antibodies, intracellular antibodies (“intramolecular antibodies”), antigen-binding fragments (such as Fv, Fab, Fab', F(ab)2, and F(ab')2), as well as single-chain antibodies (scFv), heavy-chain antibodies (such as camel antibodies), and humanized antibodies (Harlow et al., 1999, Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY; Harlow et al., 1989, Antibodies: A Laboratory Manual, Cold Spring Harbor, New York; Houston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; Bird et al., 1988, Science 242:423-426).

[0042] Full-length antibodies consist of two heavy chains and two light chains. Variable regions of the light and heavy chains are responsible for antigen binding. These variable domains of the heavy and light chains may be referred to as “VH” and “VL”, respectively. The variable regions of the two chains typically contain three highly variable loops called complementarity-determining regions (CDRs): light chain (LC) CDRs, including LC-CDR1, LC-CDR2, and LC-CDR3; heavy chain (HC) CDRs, including HC-CDR1, HC-CDR2, and HC-CDR3. The CDR boundaries of the antibody and antigen-binding fragments disclosed herein can be defined or identified according to the Kabat, Chothia, or Al-Lazikani conventions (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). The three core-residue junctions (CDRs) of either the heavy or light chain are interspersed between flanking segments called framework regions (FRs), which are more conserved than the CDRs and form a scaffold supporting the hypervariable loop. The constant regions of the heavy and light chains do not participate in antigen binding but exhibit various effector functions. Antibodies are classified according to the amino acid sequence of their heavy chain constant regions. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, characterized by the presence of α, δ, ε, γ, and μ heavy chains, respectively. Several major antibody classes are further subdivided into subclasses, such as lgG1 (γ1 heavy chain), lgG2 (γ2 heavy chain), lgG3 (γ3 heavy chain), lgG4 (γ4 heavy chain), lgA1 (α1 heavy chain), or lgA2 (α2 heavy chain).

[0043] As used herein, the term "antigen-binding fragment" refers to an antibody fragment, including, for example, bifunctional antibodies, Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized bifunctional antibodies (ds bifunctional antibodies), single-chain Fv (scFv), scFv dimers (bivalent bifunctional antibodies), multispecific antibodies formed from a portion of an antibody containing one or more CDRs, single-domain antibodies (sdAbs) (e.g., camelified single-domain antibodies), nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment that binds to an antigen but does not contain a complete antibody structure. An antigen-binding fragment is capable of binding to the same antigen with a parent antibody or a fragment of a parent antibody (e.g., a parental scFv). In some embodiments, the antigen-binding fragment may contain one or more CDRs from a specific human antibody that have been grafted into a frame region from one or more different human antibodies.

[0044] "Fv" is the smallest antibody fragment containing a complete antigen recognition and binding site. This fragment consists of a dimer of a heavy chain variable region domain and a light chain variable region domain in tight, non-covalent association. The folding of these two domains generates six hypervariable rings (three from each of the heavy and light chains), which contribute amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, although the affinity is lower than the entire binding site, even a single variable domain (or half of the Fv containing only three antigen-specific CDRs) can recognize and bind to the antigen.

[0045] A "single-chain Fv," also abbreviated as "sFv" or "scFv," is an antibody fragment containing VH and VL antibody domains linked together to form a single polypeptide chain. In some embodiments, the scFv polypeptide further includes a polypeptide linker between the VH and VL domains, which allows the scFv to form the structure required for antigen binding. For a review of scFv, see Plückthun. The Pharmacology of Monoclonal Antibodies Volume 113, edited by Rosenburg and Moore, Springer-Verlag, New York, pp. 269-315 (1994).

[0046] A basic 4-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 contain 10 antigen-binding sites, while IgA antibodies contain 2–5 basic 4-chain units that can polymerize to combine with the J chain to form a polyvalent assembly. In the case of IgG, the 4-chain unit is typically about 150,000 Daltons. Each L chain is linked to the H chain by a covalent disulfide bond, and two H chains are linked to each other by one or more disulfide bonds depending on the H chain isoform. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has a variable domain (VH) at its N-terminus, followed by three constant domains (CH) for both the α and γ chains, and four CH domains for the μ and ε isoforms. Each L chain has a variable domain (VL) at its N-terminus, followed by a constant domain at its other end. VL is aligned with VH, and CL is aligned with the first constant domain (CH1) of the heavy chain. Specific amino acid residues are thought to form the interface between the variable domains of the light chain and the variable domains of the heavy chain. VH and VL pair together to form a single antigen-binding site. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th ed., Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds.), Appleton & Lange, Norwalk, Conn., 1994, p. 71 and Chapter 6. The L chain from any vertebrate species can be designated as one of two distinct types, called κ and λ, based on the amino acid sequence of its constant domain. Immunoglobulins can be designated as different classes or isotypes based on the amino acid sequence of the heavy chain (CH) constant domain. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, each with a heavy chain designated as α, δ, ε, γ, and μ, respectively. Based on relatively minor differences in CH sequence and function, the γ and α classes are further subdivided into subclasses, such as the following subclasses expressed in humans: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0047] The Fc fragment comprises the carboxyl-terminal portions of two H chains held together by disulfide bonds. The effector function of an antibody is determined by the sequence in the Fc region, which is also recognized by Fc receptors (FcRs) found on certain types of cells.

[0048] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of either the heavy or light chain. The variable domains of the heavy and light chains are referred to as "VH" and "VL," respectively. These domains are typically the largest variable parts of an antibody (relative to other antibodies of the same class) and contain antigen-binding sites. Camelidae Antibodies of species with only heavy chains possess a single heavy chain variable region, which is called a "VHH". Therefore, VHH is a special type of VH.

[0049] The term "variability" refers to the fact that the sequences of certain segments of a variable domain differ widely between antibodies. The V domain mediates antigen binding and defines the specificity of a particular antibody for its specific antigen. However, variability is not uniformly distributed across the entire span of the variable domain. Rather, in both the light and heavy chain variable domains, variability is concentrated in three segments called hypervariable regions (HVRs). The more conserved portions of the variable domain are called framework regions (FRs). The variable domains of the natural heavy and light chains each contain four FR regions, predominantly employing a β-sheet configuration, linked by three HVRs that form loops connecting β-sheet structures and, in some cases, forming part of a β-sheet structure. The HVRs in each chain are held together closely adjacent to each other by the FR regions and, together with HVRs from the other chain, contribute to the formation of the antibody's antigen-binding site (see Kabat et al., Sequences of Immunological Interest, 5th ed., National Institute of Health, Bethesda, Md. (1991)). Constant domains do not directly participate in the binding of antibodies to antigens, but they exhibit various effector functions, such as antibody participation in antibody-dependent cytotoxicity.

[0050] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous group of antibodies, i.e., the individual antibodies constituting said group are identical except for possible small amounts of possibly naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation). Monoclonal antibodies are highly specific (targeting a single antigenic site). Unlike polyclonal antibody formulations, which typically comprise different antibodies targeting different determinants (epitopes), each monoclonal antibody targets a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous because they are synthesized via hybridoma culture and are not contaminated by other immunoglobulins. The modifier "monoclonal" indicates the characteristic of an antibody obtained from a substantially homogeneous group of antibodies and should not be construed as requiring the production of said antibody by any particular method. For example, the monoclonal antibody intended for use according to this application can be prepared by a variety of techniques, including, for example, hybridoma methods (e.g., Kohler and Milstein). Nature256:495-97 (1975); Hongo et al., Hybridoma, 14 (3): 253-260 (1995); Harlow et al., Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd edition, 1988); Hammerling et al. Monoclonal Antibodies and T - Cell Hybridomas 563-681 (Elsevier, NY, 1981), recombinant DNA methods (see, for example, U.S. Patent No. 4,816,567), phage display technology (see, for example, Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2):299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004);Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004)) and techniques for generating human antibodies or human-like antibodies in animals having some or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, for example, WO 1998 / 24893, WO 1996 / 34096, WO 1996 / 33735, WO 1991 / 10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993); Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Patent Nos. 5,545,807, 5,545,806, 5,569,825, 5,625,126, 5,633,425, and 5,661,016; Marks et al., Bio / Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813 (1994); Fishwild et al., Nature Biotechnol.14: 845-851 (1996); Neuberger, Nature Biotechnol. 14: 826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13: 65-93 (1995)).

[0051] The terms "full-length antibody," "intact antibody," or "complete antibody" are used interchangeably to refer to an antibody in essentially its complete form, rather than an antibody fragment. Specifically, full-length 4-chain antibodies include antibodies having both a heavy chain and a light chain and including an Fc region. The constant domain can be a native sequence constant domain (e.g., a human native sequence constant domain) or a variant of its amino acid sequence. In some cases, an intact antibody may have one or more effector functions.

[0052] The term "bifunctional antibody" refers to an antibody constructed by combining the VH domain with the V... L Small antibody fragments are prepared from sFv fragments (see previous paragraph) with short linkers (approximately 5-10 residues) between their domains. These linkers enable interchain rather than intrachain pairing of the V domain, resulting in bivalent fragments—fragments with two antigen-binding sites. Bispecific bifunctional antibodies are heterodimers of two “cross-linked” sFv fragments, where the VH domain and V... L The domains are located on different polypeptide chains. Bifunctional antibodies are described in detail, for example, EP 404,097; WO 93 / 11161; Hollinger et al. Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993)

[0053] Monoclonal antibodies as used in this article explicitly include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and / or light chains is identical or homologous to the corresponding sequence in an antibody derived from a specific species or belonging to a specific antibody class or subclass, while the remainder of one or more chains is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass; and fragments of such antibodies, provided they exhibit the desired biological activity (US Patent No. 4,816,567; Morrison et al.). Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). Chimeric antibodies of interest in this paper include PRIMATTZFD® antibodies, wherein the antigen-binding region of the antibody is derived from antibodies produced by immunizing macaques, for example, with the antigen of interest. As used herein, “humanized antibody” is used as a subset of “chimeric antibody”.

[0054] As used herein, the term “CDR” or “complementarity-determining region” is intended to refer to a non-continuous antigenic combination site found within the variable region of both the heavy and light chain polypeptides. These specific regions have been described in the following literature: Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., US Dept. of Health and Human Services, “Sequences of proteins of immunological interest” (1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987); Al-Lazikani B. et al., J. Mol. Biol. , 273:927-948 (1997); MacCallum et al., J. Mol. Biol. 262:732-745 (1996); Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Lefranc MP et al., Dev. Comp. Immunol. , 27: 55-77 (2003); and Honegger and Plückthun, J. Mol. Biol. , 309:657-670 (2001), where the definition includes overlap or subset of amino acid residues when compared with each other. However, the application of any definition means that the CDR of the antibody or its transplanted antibody or variant is intended to be within the scope of the terminology defined and used herein. The amino acid residues covering the CDR as defined in each of the references cited above are listed in Table A below for comparison. CDR prediction algorithms and interfaces are known in the art, including, for example, those by Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Ehrenmann F. et al., Nucleic Acids Res. , 38: D301-D307 (2010); and Adolf-Bryfogle J. et al., Nucleic Acids Res. References cited in this paragraph are incorporated herein by reference in their entirety for the purposes of this application and may be included in one or more of the claims herein.

[0055] Table A: CDR Definition

[0056] 1 Residue numbering follows the nomenclature of Kabat et al., as above.

[0057] 2 Residue numbering follows the nomenclature of Chothia et al., as above.

[0058] 3 Residue numbering follows the nomenclature of MacCallum et al., as above.

[0059] 4 Residue numbering follows the nomenclature of Lefranc et al., as above.

[0060] 5 Residue numbering follows the Honegger and Plückthun nomenclature, as described above.

[0061] The expressions “as in Kabat variable domain residue numbering” or “as in Kabat amino acid position numbering” and their variations refer to the numbering system used by Kabat et al. (ibid.) for heavy chain or light chain variable domains in antibody editing. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortenings or insertions within the FR or hypervariable region (HVR) of the variable domain. For example, a heavy chain variable domain may include a single amino acid insertion after residue 52 of H2 (according to Kabat residue 52a) and residues inserted after heavy chain FR residue 82 (e.g., according to Kabat residues 82a, 82b, and 82c, etc.). For a given antibody, the Kabat number of the residues can be determined by comparing homologous regions of the antibody sequence with “standard” Kabat numbered sequences.

[0062] Unless otherwise indicated herein, the residue numbers in the immunoglobulin heavy chain are derived from the EU index, as described by Kabat et al. (ibid.), with slight modifications. "EU index, as described by Kabat," refers to the residue numbers of human IgG1 EU antibodies.

[0063] “Frame” or “FR” residues are those variable domain residues other than the CDR residues defined herein.

[0064] As used in this article, “immunoassay” refers to any binding assay that uses antibodies capable of specifically binding to target molecules to detect and quantify the target molecules.

[0065] "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies containing a minimal sequence derived from a non-human antibody. To a large extent, humanized antibodies are human immunoglobulins (recipient antibodies) in which residues from the hypervariable region (HVR) of the receptor are replaced by residues from the hypervariable region of a non-human species (donor antibody), such as mice, rats, rabbits, or non-human primates possessing the desired antibody specificity, affinity, and capability. In some cases, the frame region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may contain residues not present in the recipient antibody or the donor antibody. These modifications are made to further improve antibody performance. Generally, humanized antibodies will contain substantially all at least one and usually two variable domains, wherein all or substantially all of the hypervariable loops correspond to the hypervariable loops of the non-human immunoglobulin, and all or substantially all of the FRs are FRs of the human immunoglobulin sequence. Humanized antibodies will also optionally contain at least a portion of the immunoglobulin constant region (Fc) (typically the constant region of human immunoglobulins). Suitable human receptor antibodies can be selected from conventional databases, such as the KABAT database, the Los Alamos database, the AbM database, and the Swiss protein database, based on homology with the nucleotide and amino acid sequences of the donor antibody. Human antibodies characterized by homology (based on amino acids) with the frame regions of the donor antibody may be suitable for providing heavy chain constant regions and / or heavy chain variable frame regions for insertion into the donor CDR. Suitable receptor antibodies capable of providing light chain constant regions or variable frame regions can be selected in a similar manner. It should be noted that the heavy and light chains of the receptor antibody do not need to originate from the same receptor antibody. Several methods for generating such humanized antibodies are described in the prior art (see, for example, EP-A-0239400 and EP-A-054951). For further details, see, for example, Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See also, for example, Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409.

[0066] "Human antibody" refers to an antibody having an amino acid sequence corresponding to that of human-produced antibodies, and / or an antibody prepared using any of the techniques disclosed herein for preparing human antibodies. This definition of human antibody explicitly excludes humanized antibodies containing non-human antigen-binding residues. Various techniques known in the art, including phage display libraries, can be used to generate human antibodies. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Cole et al. also used this method to prepare human monoclonal antibodies. Monoclonal Antibodies and Cancer Therapy Alan R. Liss, 77 (1985); Boerner et al., J. Immunol., The method described in 147(1):86-95 (1991). See also van Dijk and van deWinkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering an antigen to a transgenic animal that has been modified to produce such antibodies in response to antigen stimulation, but whose endogenous loci have been disabled, for example, by immunizing xenogeneic mice (see, for example, U.S. Patent Nos. 6,075,181 and 6,150,584 for XENOMOUSE™ technology). Also see, for example, Li et al., for human antibodies produced via human B-cell hybridoma technology. Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006).

[0067] The term "donor antibody" refers to an antibody (monoclonal antibody and / or recombinant antibody) that contributes its variable region, CDR or other functional fragment or analogue of amino acid sequence to a first immunoglobulin conjugate, thereby giving the altered immunoglobulin coding region and the altered antibody expressed therefrom the antigen specificity and neutralizing activity characteristic of the donor antibody.

[0068] The term "receptor antibody" refers to an antibody (monoclonal antibody and / or recombinant antibody) that is heterologous to a donor antibody, which contributes all (or any portion, but in some embodiments, all) of its amino acid sequence encoding its heavy chain and / or light chain framework region and / or its heavy chain and / or light chain constant region to the first immunoglobulin conjugate. In some embodiments, the human antibody is a receptor antibody.

[0069] As used herein, the terms “attach,” “attached,” “fuse,” or “fused” refer to the connection or union of two or more components by bonds, linkages, forces, or ligations, including direct or indirect connections, thereby, for example, directly binding a first polypeptide to a second polypeptide or material, and, for example, placing one or more intermediate compounds (e.g., amino acids, peptides, polypeptides, etc.) between the first polypeptide and the second polypeptide or material.

[0070] The "percentage of amino acid sequence identity (%)" or "homology" for the peptide and antibody sequences identified herein is defined as the percentage of amino acid residues in the candidate sequence that are identical to amino acid residues in the compared peptide, after sequence alignment and taking into account any conserved substitutions as part of sequence identity. The alignment used to determine the percentage of amino acid sequence identity can be performed in various ways within the capabilities of those skilled in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. Those skilled in the art can determine appropriate parameters for measuring the alignment, including any algorithms required to achieve maximum alignment across the full length of the compared sequences. However, for the purposes of this document, the amino acid sequence identity value % (Edgar, RC, ...) is generated using the sequence comparison computer program MUSCLE. Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, R.C., BMC Bioinformatics 5(1):113, 2004).

[0071] "Homologous" refers to the sequence similarity or identity between two polypeptides or two nucleic acid molecules. Two compared sequences are homologous when positions at those positions are occupied by the same base or amino acid monomer subunit. For example, if a position in each of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percentage of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions being compared, multiplied by 100. For example, if six out of ten positions in two sequences are matching or homologous, then the two sequences are 60% homologous. For instance, the DNA sequences ATTGCC and TATGGC share 50% homology. Generally, comparisons are made when two sequences are aligned to give the maximum homology.

[0072] The term "constant domain" refers to a portion of an immunoglobulin molecule that has a more conserved amino acid sequence compared to another part of the immunoglobulin (the variable domain, which contains the antigen-binding site). The constant domain contains the C16 heavy chain. H1. C H 2 and C H 3. Structural domains (collectively referred to as C) H ) and light chain CHL (or C L ) structural domain.

[0073] The “light chains” of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two distinct types (called kappa (“κ”) and lamuda (“λ”)) based on the amino acid sequence of their constant domains.

[0074] The “CH1 domain” (also known as the “C1” of the “H1” domain) typically extends from about amino acid 118 to about amino acid 215 (EU numbering system).

[0075] The "hinge region" is generally defined as the region in IgG corresponding to Glu216 to Pro230 of human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). The hinge region of other IgG isotypes can be aligned with the IgG1 sequence by placing the first and last cysteine ​​residues that form the inter-heavy chain SS bond in the same position.

[0076] The "CH2 domain" (also known as the "C2 domain") of the human IgG Fc region typically extends from about amino acid 231 to about amino acid 340. The unique feature of the CH2 domain is that it does not pair tightly with another domain. Instead, two N-linked branched carbohydrate chains are intercalated between the two CH2 domains of the intact native IgG molecule. It is speculated that the carbohydrates provide an alternative to domain-domain pairing and contribute to the stability of the CH2 domain. (Burton) Molec Immunol. 22:161-206 (1985).

[0077] The “CH3 domain” (also known as the “C3 domain”) contains a chain segment from the C-terminus of the Fc domain to the CH2 domain (i.e., from approximately amino acid residue 341 of the antibody sequence to the C-terminus, typically at amino acid residue 446 or 447 of IgG).

[0078] The term "Fc domain" or "fragment crystallizable region" is used herein to define the C-terminal region of the immunoglobulin heavy chain, including both native sequence Fc domains and variant Fc domains. Although the boundaries of immunoglobulin heavy chain Fc domains may vary, the human IgG heavy chain Fc domain is generally defined as the segment extending from an amino acid residue at position Cys226 or from position Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc structure (residue 447 according to the EU numbering system) can be removed, for example, during antibody production or purification or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Therefore, compositions of complete antibodies may comprise antibody populations with all K447 residues removed, antibody populations without K447 residues removed, and antibody populations having mixtures with or without K447 residues. Suitable native sequence Fc domains for the antibodies described herein include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4.

[0079] As used herein, the term "epitope" refers to a specific set of atoms or amino acids bound to an antigen by an antibody or antibody moiety. If two antibodies or antibody moiety exhibit competitive binding to an antigen, they can bind to the same epitope within the antigen.

[0080] As used herein, when a first antibody or fragment thereof inhibits the binding of a second antibody or fragment thereof to the target antigen by at least about 50% (e.g., at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) in the presence of an equimolar concentration of the first antibody or fragment thereof, the first antibody or fragment thereof “competes” with the second antibody or fragment thereof for binding to the target antigen, or vice versa. A high-throughput method for “binding” antibodies based on antibody cross-competition is described in PCT Publication No. WO 03 / 48731.

[0081] As used herein, the terms “specifically bind,” “specifically recognize,” and “specific to” refer to measurable and reproducible interactions, such as the binding between a target and an antibody or antibody moiety, which determines the presence of the target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody or antibody moiety that specifically recognizes a target (which may be an epitope) binds to that target with greater affinity, cohesion, ease of binding, and / or longer duration of binding than it does to other targets. In some embodiments, the degree to which an antibody binds to an irrelevant target is less than about 10% of the antibody's binding to the target, as measured, for example, by radioimmunoassay (RIA). In some embodiments, antibodies that specifically bind to a target have ≤10% binding. -5 M, ≤10 -6 M, ≤10 -7M, ≤10 -8 M, ≤10 -9 M, ≤10 -10 M, ≤10 -11 M or ≤10 -12 The dissociation constant of M (K) D In some embodiments, the antibody specifically binds to an epitope on a protein that is conserved across proteins from different species. In some embodiments, specific binding may include, but is not required to be, exclusive binding. The binding specificity of the antibody or antigen-binding domain can be determined experimentally using methods known in the art. Such methods include, but are not limited to, Western blotting, ELISA assays, RIA assays, ECL assays, IRMA assays, EIA assays, BIACORE™ assays, and peptide scanning.

[0082] The term "specificity" refers to the selective recognition of a specific epitope of an antigen by an antigen-binding protein or antibody. For example, natural antibodies are monospecific. As used herein, the term "multispecific" means that an antigen-binding protein or antibody has two or more antigen-binding sites, wherein at least two antigen-binding sites bind to different antigens or different epitopes of the same antigen. As used herein, "bispecific" means that an antigen-binding protein or antibody has two different antigen-binding specificities. As used herein, the term "monospecific" antibody means an antibody having one or more binding sites, wherein each binding site binds to the same epitope of the same antigen.

[0083] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its bound collateral (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the inherent binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its collateral Y is generally expressed as a dissociation constant (Kd). Affinity can be measured by commonly used methods known in the art, including those described herein. Low-affinity antibodies typically bind antigens slowly and tend to dissociate readily, while high-affinity antibodies typically bind antigens more quickly and tend to maintain binding for longer periods. Various methods for measuring binding affinity are known in the art, any of which can be used for the purposes of this application. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0084] As used in this article, "on-rate", "rate of association", "association rate" or "k" on "It can also be determined using methods such as biolayer interferometry and surface plasmon resonance (SPR), as mentioned above."

[0085] "Isolated" antibodies are antibodies (e.g., natural or recombinant) that have been identified, separated, and / or recovered from components of their production environment. Preferably, the isolated peptide does not associate with any other components from its production environment. Contaminant components of its production environment, such as those produced by recombinantly transfected cells, are substances that typically interfere with the research, diagnostic, or therapeutic use of the antibody and may include enzymes, hormones, and other proteins or non-protein solutes. In a preferred embodiment, the peptide will: (1) be purified to a greater than 95% by weight of antibody, and in some embodiments, greater than 99% by weight of antibody, as determined by, for example, the Lowry method; (2) be purified to a degree sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a twist-cup sequencer; or (3) be purified to a degree determined to be homogeneous by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver staining. Isolated antibodies include in situ antibodies from recombinant cells, since at least one component of the antibody's native environment will be absent. However, isolated peptides or antibodies are typically prepared through at least one purification step.

[0086] An "isolated" nucleic acid molecule encoding the constructs, antibodies, or antigen-binding fragments described herein is a nucleic acid molecule that is identified and isolated from at least one contaminant nucleic acid molecule typically associated with it in the environment in which it is produced. Preferably, the isolated nucleic acid is not associated with any components related to the environment in which it is produced. The isolated nucleic acid molecule encoding the polypeptides and antibodies described herein is in a form different from the form or environment in which it is found in nature. Therefore, the isolated nucleic acid molecule is different from the nucleic acids naturally present in cells that encode the polypeptides and antibodies described herein. Isolated nucleic acids include nucleic acid molecules typically found in cells containing nucleic acid molecules, but which are located extrachromosomally or at chromosomal locations other than their natural chromosomal locations.

[0087] Nucleic acids are "operably linked" when they are placed in a functional relationship with another nucleic acid sequence. For example, if the DNA of a pre-sequence or secretory leader sequence is expressed as a pre-protein involved in polypeptide secretion, then the DNA of the pre-sequence or secretory leader sequence is operably linked to the DNA of the polypeptide; if a promoter or enhancer affects the transcription of a coding sequence, then the promoter or enhancer is operably linked to that sequence; or, if a ribosome binding site is positioned to facilitate translation, then the ribosome binding site is operably linked to the coding sequence. Generally, "operably linked" means that the linked DNA sequences are contiguous, and in the case of a secretory leader sequence, they are contiguous and located within the reading frame. However, enhancers do not necessarily have to be contiguous. Ligation is accomplished by connecting at a convenient restriction site. If these sites are not present, synthetic oligonucleotide adaptors or linkers are used according to conventional practice.

[0088] "Separated" refers to a change from or separation from the native state. For example, nucleic acids or peptides that are naturally present in the native state in a living subject are not "separated," but the same nucleic acids or peptides that are partially or completely separated from their native coexisting substances are "separated." Separated nucleic acids or proteins can exist in a substantially purified form or in non-native environments, such as host cells.

[0089] As used herein, the term "hybridoma" refers to a cell resulting from the fusion of B lymphocytes with a fusion pair (such as myeloma cells). Hybridoma cells can be cloned and maintained indefinitely in cell cultures and are capable of producing monoclonal antibodies. Hybridoma cells can also be considered hybrid cells.

[0090] The terms “nucleic acid molecule,” “nucleic acid,” and “polynucleotide” are used interchangeably and refer to polymers of nucleotides. Such nucleotide polymers may contain natural and / or non-natural nucleotides and include, but are not limited to, DNA, RNA, and PNA. “Nucleic acid sequence” refers to a linear sequence of nucleotides containing a nucleic acid molecule or polynucleotide. “Isolated nucleic acid” refers to a nucleic acid segment or fragment that is isolated from the sequence sidebanded therewith in its natural state, i.e., a DNA fragment removed from a sequence typically adjacent to the segment (i.e., a sequence adjacent to a segment in its naturally occurring genome). The term also applies to nucleic acids that have been substantially purified from other components naturally accompanying the nucleic acid (i.e., RNA, DNA, or protein) that naturally accompany the nucleic acid in cells. Therefore, the term includes, for example, recombinant DNA incorporated into vectors, autonomously replicating plasmids or viruses, or into the genomic DNA of prokaryotes or eukaryotes, or recombinant DNA existing as a separate molecule independent of other sequences (i.e., cDNA or genomic or cDNA fragments produced by PCR or restriction enzyme digestion). It also includes recombinant DNA, which is part of a hybrid gene encoding an additional polypeptide sequence.

[0091] As used herein, “complementarity” refers to nucleic acids, and is a broad concept of sequence complementarity between regions of two nucleic acid strands or between two regions of the same nucleic acid strand. It is known that adenine residues in a first nucleic acid region can form specific hydrogen bonds (“base pairing”) with residues in a second nucleic acid region, and if said residues are thymine or uracil, then the second nucleic acid region is antiparallel to the first region. Similarly, it is known that cytosine residues in the first nucleic acid strand can base pair with residues on a second nucleic acid strand antiparallel to the first strand, provided that said residues are guanine. If a first region of a nucleic acid is arranged antiparallel to a second region of the same or different nucleic acid, and at least one nucleotide residue in the first region can base pair with a residue in the second region, then the two regions are complementary. In some embodiments, the first region comprises a first portion, and the second region comprises a second portion, such that when the first and second portions are arranged antiparallel, at least about 50% and / or at least about 75%, or at least about 90%, or at least about 95% of the nucleotide residues in the first portion can base pair with nucleotide residues in the second portion. In some implementations, all nucleotide residues in the first part can be base-paired with nucleotide residues in the second part.

[0092] As used herein, the term "vector" refers to a nucleic acid molecule capable of proliferating another nucleic acid to which it is linked. This term includes vectors in the form of self-replicating nucleic acid structures as well as vectors incorporated into the genome of a host cell. Vectors can be plasmids, bacteriophages, bacterial artificial chromosomes, or yeast artificial chromosomes. Vectors can be DNA vectors or RNA vectors. Vectors can be autonomously replicating extrachromosomal vectors or vectors integrated into the host genome. Some vectors are capable of guiding the expression of the nucleic acids to which they are operatively linked. Such vectors are referred to herein as "expression vectors."

[0093] "Encoding" refers to the inherent characteristic of a specific nucleotide sequence in a polynucleotide (such as a gene, cDNA, or mRNA) that serves as a template for the synthesis of other polymers and macromolecules having a defined nucleotide sequence (i.e., rRNA, tRNA, and mRNA) or a defined amino acid sequence in biological processes, and the resulting biological characteristics. Therefore, if the transcription and translation of mRNA corresponding to a gene produces a protein in a cell or other biological system, then the gene encodes that protein. The coding strand, whose nucleotide sequence is identical to the mRNA sequence and is generally provided in the sequence listing, and the non-coding strand, which serves as a transcription template for a gene or cDNA, can both be referred to as encoding the protein or other product of that gene or cDNA.

[0094] The terms "polypeptide" and "peptide" are used interchangeably and refer to a polymer of amino acid residues, and are not limited to a minimum length. Such a polymer of amino acid residues can contain native or non-native amino acid residues. Full-length proteins and fragments thereof are included in this definition. The term also includes post-expression modifications of the polypeptide, such as glycosylation, sialylation, acetylation, phosphorylation, etc. Furthermore, "polypeptide" includes modifications to the native sequence, such as deletions, additions, and substitutions (often conserved), provided that the polypeptide retains the desired activity. These modifications can be intentional, such as by site-directed mutagenesis, or can be accidental, such as by mutations in the host producing the protein or errors caused by PCR amplification.

[0095] As used in this article, “combination” refers to the covalent connection between one molecule and another.

[0096] As used herein, the term "variant" is a nucleic acid or peptide sequence that differs from a reference nucleic acid or peptide sequence, respectively, but retains the essential biological characteristics of the reference molecule. Sequence changes in nucleic acid variants may not alter the amino acid sequence of the peptide encoded by the reference nucleic acid, or may result in substitutions, additions, deletions, fusions, or truncations of amino acids. Sequence changes in peptide variants are generally limited or conserved, so the sequences of the reference peptide and the variant are very similar overall and identical in many regions. The amino acid sequences of the variant and the reference peptide may differ due to any combination of one or more substitutions, additions, or deletions. Variants of nucleic acids or peptides can be naturally occurring variants, such as allelic variants, or can be variants known not to exist naturally. Non-naturally occurring variants of nucleic acids and peptides can be prepared by mutagenesis or direct synthesis. In various embodiments, the variant sequence is at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 88%, at least 87%, at least 86%, or at least 85% identical to the reference sequence.

[0097] As described in this article by Spree USA. The term "regulation" can refer to any method that alters the level or activity of a substrate. Non-limiting examples of protein regulation include affecting expression (including transcription and / or translation), affecting folding, affecting degradation or protein turnover, and affecting protein localization. Non-limiting examples of enzyme regulation include affecting enzyme activity. A "regulator" is a molecule whose activity includes affecting the level or activity of a substrate. Regulators can be direct or indirect. Regulators can be used to activate, inhibit, or otherwise regulate their substrates.

[0098] The term "control sequence" refers to the DNA sequence required to express an operable, linked coding sequence in a specific host organism. Control sequences suitable for prokaryotes include, for example, promoters, optional operon sequences, and ribosome binding sites. Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.

[0099] "Pharmaceutically acceptable carriers" refer to conventional, non-toxic solid, semi-solid, or liquid fillers, diluents, encapsulating materials, formulation aids, or carriers used in conjunction with therapeutic agents to form a "pharmaceutical composition" for administration to a subject. Pharmaceutically acceptable carriers are non-toxic to the recipient at the doses and concentrations used and are compatible with other components in the formulation. Pharmaceutically acceptable carriers are suitable for the formulations used.

[0100] The “diluent” of interest herein is pharmaceutically acceptable (safe and non-toxic for human administration) and can be used to prepare liquid formulations, such as lyophilized and reconstituted formulations. Exemplary diluents include sterile water, water for sterile injection (BWFI), pH buffer solutions (e.g., phosphate-buffered saline), sterile saline solutions, Ringer's solution, or dextran solution. In an alternative embodiment, the diluent may comprise an aqueous solution of salt and / or buffer.

[0101] A "preservative" is a compound that can be added to the formulations described herein to reduce bacterial activity. Adding preservatives can, for example, facilitate the manufacture of multi-purpose (multi-dosage) formulations. Examples of potential preservatives include octadecyl dimethylbenzyl ammonium chloride, hexamethyl diammonium chloride, benzalkonium chloride (a mixture of alkylbenzyl dimethyl ammonium chlorides, where the alkyl group is a long-chain compound), and benzyl chloride. Other types of preservatives include aromatic alcohols such as phenol, butanol, and benzyl alcohol; alkyl hydroxybenzoates such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol. The most commonly used preservative in this article is benzyl alcohol.

[0102] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to preparations that are in a form that enables the biological activity of one or more active ingredients and do not contain any additional components that would cause unacceptable toxicity to a subject to whom the formulation will be administered. Such formulations may be sterile.

[0103] "Sterile" preparations are sterile or substantially free of live microorganisms and their spores.

[0104] "Stable" formulations are those in which proteins substantially retain their physical and chemical stability and integrity during storage. Various analytical techniques exist in the art for measuring protein stability, and a review of these techniques can be found in [link to relevant documentation]. Peptide and Protein Drug Delivery, 247-301, Vincent Lee (ed.), Marcel Dekker, Inc., New York, NY, Pubs. (1991); and Jones, A. Adv. Drug Delivery Review.10: 29-90 (1993). Stability can be measured at a selected temperature and over a selected time period. For rapid screening, formulations can be kept at 40°C for 2 weeks to 1 month and then their stability can be measured. If a formulation is to be stored at 2-8°C, it should generally be stable at 30°C or 40°C for at least 1 month, and / or stable at 2-8°C for at least 2 years. If a formulation is to be stored at 30°C, it should generally be stable at 30°C for at least 2 years, and / or stable at 40°C for at least 6 months. For example, the degree of aggregation during storage can be used as an indicator of protein stability. Thus, a “stable” formulation can be one in which the amount of protein present in aggregate form is less than about 10%, and preferably less than about 5%. In other embodiments, any increase in aggregate formation during formulation storage can be identified.

[0105] A "reconstituted" formulation is a formulation prepared by dissolving a lyophilized protein or antibody formulation in a diluent, thereby dispersing the protein as a whole. Reconstituted formulations are suitable for administration (e.g., subcutaneous injection) to patients treated with the protein of interest, and in some embodiments, may be formulations suitable for parenteral or intravenous administration.

[0106] An "isotonic" formulation is one whose osmotic pressure is substantially the same as that of human blood. The osmotic pressure of an isotonic formulation is generally between approximately 250 and 350 mOsm. The term "hypotonic" describes a formulation with an osmotic pressure lower than that of human blood. Correspondingly, the term "hypertonic" is used to describe a formulation with an osmotic pressure higher than that of human blood. For example, isotonicity can be measured using a vapor pressure osmoremeter or a cryo-osmoremeter. The formulations in this application can be made hypertonic by adding salts and / or buffer solutions.

[0107] “ Recombinant AAV vector (rAAV vector)A recombinant adeno-associated virus (rAAV) vector refers to a polynucleotide vector containing one or more heterologous sequences (i.e., nucleic acid sequences not derived from AAV), which are side-joined with at least one and, in some embodiments, two AAV inverted terminal repeat (ITR) sequences. Such an rAAV vector, when present in a host cell infected with a helper virus (or expressing a helper function) and expressing the AAV rep and cap gene products (i.e., the AAV Rep and Cap proteins), can replicate and be packaged into infectious viral particles. When the rAAV vector is incorporated into a larger polynucleotide (e.g., into a chromosome or another vector, such as a plasmid for cloning or transfection), the rAAV vector can be referred to as a "pre-vector," which is "rescued" by replication and encapsulation in the presence of AAV packaging and helper functions. The rAAV vector can take many forms, including but not limited to plasmids, linear artificial chromosomes, lipid complexes, encapsulation in liposomes, and capsidation within viral particles, particularly AAV particles. The rAAV vector can be packaged into the capsid of an AAV virus to produce "recombinant adeno-associated virus particles (rAAV particles)."

[0108] “ AAV inverted terminal repeat (ITR) "Sequence" is a well-known term in the art; it is a sequence of approximately 145 nucleotides located at both ends of a naturally occurring single-stranded AAV genome. The outermost 125 nucleotides of the ITR can be oriented in two different ways, resulting in heterogeneity between different AAV genomes and between the ends of a single AAV genome. The outermost 125 nucleotides also contain several shorter self-complementary regions (called A, A', B, B', C, C', and D regions) that allow intrastrand base pairing to occur within this portion of the ITR.

[0109] As used herein, the terms “transfection” or “transformation” or “transduction” refer to the process of transferring or introducing exogenous nucleic acids into host cells. “Transfected” or “transformed” or “transduced” cells are cells that have been transfected, transformed, or transduced with exogenous nucleic acids. The cells include primary subject cells and their progeny.

[0110] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells in which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include “transformers” and “transformed cells,” which include primary transformed cells and their derived progeny, regardless of the number of passages. The nucleic acid content of progeny cells may not be identical to that of the parent cells, and they may contain mutations. This article includes screened or selected mutant progeny that have the same function or biological activity as the initially transformed cells.

[0111] As used herein, “treatment” or “treating” is a method for obtaining a beneficial or desired outcome (including clinical outcomes). For the purposes of this application, a beneficial or desired clinical outcome includes, but is not limited to, one or more of the following: relief of one or more symptoms caused by a disease, reduction of the severity of the disease, stabilization of the disease (e.g., prevention or delay of disease exacerbation), prevention or delay of disease spread, prevention or delay of disease recurrence, delay or slowing of disease progression, improvement of disease status, provision of disease remission (partial or complete remission), reduction of the dosage of one or more other medications required to treat the disease, delay of disease progression, improvement or enhancement of quality of life, increase in weight gain, and / or prolongation of survival. The methods of this application cover any one or more of these treatment aspects.

[0112] As used herein, the terms “effective amount” and “pharmaceutically effective amount” refer to an amount of a pharmaceutical agent sufficient to provide the desired biological outcome. This outcome may be a reduction (e.g., a reduction of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100%) and / or an alleviation of the signs, symptoms, or cause of a disease or condition, or any other desired alteration of a biological system.

[0113] The terms "inhibition" or "inhibit" refer to a reduction or cessation of any phenotypic trait, or a reduction or cessation of the occurrence, extent, or likelihood of that trait. "Reduction" or "inhibit" means a decrease, reduction, or cessation of activity, function, and / or quantity compared to a reference. In some embodiments, "reduction" or "inhibit" means an overall reduction of 20% or greater (e.g., at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100%). In another embodiment, "reduction" or "inhibit" means an overall reduction of 50% or greater. In yet another embodiment, "reduction" or "inhibit" means an overall reduction of 75%, 85%, 90%, 95%, or greater.

[0114] As used herein, “delayed disease development” means postponing, hindering, slowing, delaying, stabilizing, suppressing, and / or postponing the development of a disease. This delay can vary in length depending on the individual’s medical history and / or the treatment received. As will be apparent to those skilled in the art, adequate or significant delay can effectively encompass prevention, as the individual does not develop the disease.

[0115] As used in this article, “prevention” includes providing protection against the onset or recurrence of disease in individuals who may be susceptible to the disease but have not yet been diagnosed with it.

[0116] As used herein, "suppression" function or activity is a decrease in function or activity when compared to conditions that are otherwise identical except for the condition or parameter of interest, or alternatively, when compared to another condition. For example, an antibody that suppresses tumor growth reduces the rate of tumor growth compared to the absence of the antibody.

[0117] As used herein, “reference” means any sample, standard, or level used for comparative purposes. References may be obtained from healthy and / or disease-free samples. In some instances, references may be obtained from untreated samples. In some instances, references may be obtained from disease-free or untreated samples of individuals. In some instances, references may be obtained from one or more healthy individuals who are not individuals or patients.

[0118] As used herein, the terms “patient,” “subject,” “individual,” etc., are used interchangeably and refer to any animal with a complement system, which in some embodiments is a mammal and in some embodiments is a human, including those who require treatment or are susceptible to a disease or its sequelae. An individual may include, for example, dogs, cats, pigs, cattle, sheep, goats, horses, rats, monkeys, mice, and humans. In some embodiments, the individual is a human.

[0119] The "human shared framework" or "recipient human framework" represents a series of human immunoglobulin V... L or V H The framework sequence is a framework of the most frequently occurring amino acid residues. Typically, this sequence is found in human immunoglobulin V. L or V H The sequence is a subgroup derived from sequences of variable structural domains. Generally, this subgroup is those described by Kabat et al. Sequences of Proteins of Immunological Interest Subgroups in, 5th edition, Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Examples include for V L The subgroup can be κ I, κ II, κ III, or κ IV, as described in Kabat et al. above. Furthermore, for V... HThe subgroup can be Subgroup I, Subgroup II, or Subgroup III as described by Kabat et al. Alternatively, the human common framework can be derived from the above, wherein specific residues are selected based on their homology with the donor framework, for example, by comparing the donor framework sequence with a collection of various human framework sequences. The recipient human framework "derived from" the human immunoglobulin framework or the human common framework may contain its same amino acid sequence, or it may contain pre-existing amino acid sequence variations. In some embodiments, the number of pre-existing amino acid variations is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less.

[0120] Affinity-matured antibodies are antibodies that have one or more modifications in one or more CDRs, such that the antibody's affinity for the antigen is improved compared to a parent antibody without these modifications. In some embodiments, affinity-matured antibodies have nanomolar or even picomolar affinity for the target antigen. Affinity-matured antibodies are produced by procedures known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describes the production of antibodies via V... H Domain and V L Domain recombination leads to affinity maturation. Random mutagenesis of CDRs and / or framework residues is described, for example, by Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al., Gene 169:147-155 (1995); Yelton et al., J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol . 154(7):3310-9 (1995); and Hawkins et al., J. Mal. Biol 226:889-896 (1992).

[0121] As used herein, the term "valence" indicates the presence of a specified number of binding sites in an antigen-binding protein. For example, a natural antibody or a full-length antibody has two binding sites and is divalent. Therefore, the terms "trivalent," "tetravalent," "pentavalent," and "hexavalent" indicate the presence of two, three, four, five, and six binding sites in an antigen-binding protein, respectively.

[0122] "Antibody effector function" refers to those biological activities attributable to the antibody's Fc region (natural sequence Fc region or amino acid sequence variant Fc region) and varies with antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation. "Reduced or minimized" antibody effector function means a reduction of at least 50% (or 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%) in effector function compared to wild-type or unmodified antibodies. The determination of antibody effector function is readily determined and measured by those skilled in the art. In a preferred embodiment, effector functions such as complement binding, complement-dependent cytotoxicity, and antibody-dependent cytotoxicity are all affected. In some embodiments, effector function is eliminated by removing mutations in glycosylation from the constant region, such as "effect-free mutations." On the one hand, an effect-free mutation is a C1q binding mutation. H N297A or DANA mutations (D265A+N297A) in region 2. Shields et al. J. Biol. Chem 276 (9): 6591-6604 (2001). Alternatively, other mutations that lead to reduced or eliminated effector function include K322A and L234A / L235A (LALA). Alternatively, effector function can be reduced or eliminated by manufacturing techniques, for example, by expression in host cells that do not undergo glycosylation (e.g., *E. coli*), or by expression in host cells that induce alterations in glycosylation patterns to be ineffective or less effective in promoting effector function (e.g., Shinkawa et al.). J. Biol. Chem . 278(5): 3466-3473 (2003)).

[0123] Antibody-dependent cell-mediated cytotoxicity (ADCC) refers to a form of cytotoxicity in which secreted Ig binding to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) enables these cytotoxic effector cells to specifically bind to target cells carrying antigens and subsequently kill the target cells with cytotoxins. Antibodies "arm" cytotoxic cells and are required to kill target cells through this mechanism. NK cells, the main cells mediating ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. Fc expression on hematopoietic cells is summarized in Ravetch and Kinet. Annu. Rev. ImmunolTable 2 on page 464 of 9:457-92 (1991). To assess the ADCC activity of the molecule of interest, in vitro ADCC assays can be performed, such as those described in U.S. Patent Nos. 5,500,362 or 5,821,337. Effector cells that can be used for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, in vivo, such as in animal models, e.g., Clynes et al., PNAS USA The ADCC activity of molecules of interest was evaluated in animal models disclosed in 95:652-656 (1998).

[0124] Effector cells are leukocytes that express one or more FcRs and perform effector functions. Specifically, effector cells express at least FcγRIII and perform ADCC effector functions. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils. Effector cells can be derived from natural sources, such as blood. Effector cells are typically lymphocytes associated with the effector phase and are used to produce cytokines (helper T cells), kill infected pathogens (cytotoxic T cells), or secrete antibodies (differentiated B cells).

[0125] Complement-dependent cytotoxicity (CDC) refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is triggered by the binding of the first component of the complement system (C1q) to an antibody (an antibody of the appropriate subclass) that binds to a homologous antigen. To assess complement activation, one can, for example, use antibodies such as those from Gazzano-Santoro et al. J. Immunol. Methods The CDC assay is described in 202:163 (1996). Peptide variants with altered Fc region amino acid sequences and increased or decreased C1q binding capacity are described in U.S. Patent Nos. 6,194,551B1 and WO99 / 51642. The contents of those patents are expressly incorporated herein by reference. See also Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0126] Half-maximum inhibitory concentration (IC50) 50 IC50 is an indicator of the effectiveness of a substance (such as an antibody) in inhibiting a specific biological or biochemical function. It indicates how much of a particular drug or other substance (inhibitor, such as an antibody) is needed to inhibit a given biological process by half. These values ​​are typically expressed as molar concentration. 50 "EC" with agonist drugs or other substances (such as antibodies) 50 Similar. EC 50This also represents the plasma concentration required to achieve 50% of its maximum effect in vivo. As used in this article, "IC50" 50 "Used to indicate the effective antibody concentration required to neutralize 50% of the antigen's biological activity in vitro. IC" 50 or EC 50 Measurements can be performed using bioassays, such as FACS analysis to inhibit ligand binding (competitive binding assay), cell-based cytokine release assays, or amplified luminescent proximity homogeneous assays (AlphaLISA).

[0127] As used herein, the “low pH dissociation factor” is defined as the percentage of antibody that dissociates from the antigen at 25°C and pH 5.8, where the antibody has been pre-bound to the antigen at pH 7.4. The low pH dissociation factor can be measured by associating the antibody and antigen (e.g., an anti-human FD antibody construct and human factor D) at pH 7.4 for 600 seconds, followed by dissociation in a buffer at pH 5.8 for 600 seconds, and calculating the percentage of antibody that dissociates from the antigen at pH 5.8. The “neutral pH dissociation factor” is defined as the percentage of antibody that dissociates from the antigen at 25°C and pH 7.4, where the antibody has been pre-bound to the antigen at pH 7.4. The neutral pH dissociation factor can be measured by associating the antibody and antigen at pH 7.4 for 600 seconds, followed by dissociation in a buffer at pH 7.4 for 600 seconds, and calculating the percentage of antibody that dissociates from the antigen at pH 7.4. Antibody-antigen association and dissociation can be measured using a variety of methods within the scope of the art, such as using biolayer interferometry.

[0128] The term "Fc receptor" or "FcR" describes a receptor that binds to the Fc domain of an antibody. Preferred FcRs are naturally occurring human FcRs. Furthermore, preferred FcRs are receptors that bind IgG antibodies (γ receptors) and include receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternative splicing forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences and are distinguished primarily by their cytoplasmic domains. The activating receptor FcγRIIA contains an activation motif (ITAM) based on the immunoreceptor tyrosine residue in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an inhibitory motif (ITIM) based on the immunoreceptor tyrosine residue in its cytoplasmic domain. (See M. Daëron, Annu. Rev. Immunol. 15:203-234 (1997)). FcR is a comprehensive review by Ravetch and Kinet. Annu. Rev. Immunol.9: 457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126: 330-41 (1995). Other FcRs, including those to be identified in the future, are covered by the term "FcR" in this paper.

[0129] The term "Fc receptor" or "FcR" also includes the neonatal receptor FcRn, which is responsible for transferring maternal IgG to the fetus. Guyer et al., J. Immunol. 117: 587 (1976); and Kim's people, J. Immunol 24: 249 (1994). Methods for measuring the binding of FcRn are known (see, for example, Ghetie and Ward, Immunol. Today 18:(12): 592-8 (1997); Ghetie et al., Nature Biotechnology 15 (7): 637-40 (1997); Hinton et al., J. Biol. Chem 279 (8): 6213-6 (2004); WO 2004 / 92219 (Hinton et al.)). The binding of human FcRn high-affinity binding peptides to FcRn in vivo and their serum half-life can be determined, for example, in transgenic mice expressing human FcRn or in transfected human cell lines, or in primates administered peptides with variant Fc regions. WO 2004 / 42072 (Presta) describes antibody variants with improved or weakened binding to FcR. See also, for example, Shields et al. J. Biol. Chem . 9(2): 6591-6604 (2001).

[0130] It should be understood that the implementation schemes described herein include “consisting of each implementation scheme” and / or “substantially consisting of each implementation scheme”.

[0131] This document uses the term "about" to refer to a value or parameter, which includes (and describes) variations relating to said value or parameter itself. For example, a description of "about X" includes a description of "X".

[0132] As used herein, the reference to "not" a certain value or parameter generally means and describes "other than a certain value or parameter". For example, "the method is not used to treat type X disease" means that the method is used to treat types of diseases other than X.

[0133] The term “about XY” as used in this article has the same meaning as “about X to about Y”.

[0134] Unless the context clearly specifies otherwise, the singular forms “a,” “or,” and “the” used herein and in the appended claims include multiple indicators.

[0135] II. Anti-Factor D Antibody Construct

[0136] In some aspects, the present invention provides isolated antibody constructs (anti-human FD antibody constructs) comprising an antibody moiety that specifically binds to human factor D (an anti-human FD antibody moiety, such as any anti-human FD antibody moiety described herein). In some embodiments, the anti-human FD antibody construct consists substantially of (or is composed of) an anti-human FD antibody moiety. Therefore, in some embodiments, an anti-human FD antibody moiety is also provided. In some embodiments, the isolated anti-human FD antibody construct has any of the characteristics and activities of the anti-human FD antibody moiety described herein.

[0137] In some embodiments, the isolated anti-human FD antibody construct further comprises a fusion conjugate (e.g., a protein or peptide). In some embodiments, the fusion conjugate is fused directly to the anti-human FD antibody moiety. In some embodiments, the fusion conjugate is fused to the anti-human FD antibody moiety via a linker (e.g., a peptide linker). In some embodiments, the fusion conjugate is an effector protein. An effector protein can be any protein that produces a specific response to a stimulus, such as altering one or more cellular functions of a target cell. In some embodiments, the fusion conjugate is an antibody moiety, such as an antibody moiety that specifically binds to a non-FD epitope.

[0138] In some embodiments, the isolated anti-human FD antibody construct is monospecific. In some embodiments, the isolated anti-human FD antibody construct is multispecific (e.g., bispecific). In some embodiments, the isolated anti-human FD antibody construct is multivalent and monospecific. In some embodiments, the isolated anti-human FD antibody construct is multivalent and multispecific (e.g., bispecific).

[0139] In some embodiments, the isolated anti-human FD antibody construct further comprises a second antibody moiety (e.g., scFv, Fab, sdAb, or full-length antibody) that specifically recognizes components of the complement pathway. In some embodiments, the second antibody moiety specifically recognizes FD. Therefore, in some embodiments, the isolated anti-human FD antibody construct comprises two or more anti-human FD antibody moieties (e.g., scFv), such as anti-human FD antibody moieties linked in tandem. In some embodiments, the second antibody moiety specifically recognizes non-FD components in the complement pathway, such as C2 or C5.

[0140] Anti-human FD antibody part

[0141] The terms "anti-human FD antibody moiety" and "anti-FD antibody moiety" are used interchangeably in this document. The use of "anti-human FD" does not exclude antibody moiety capable of binding to human FD or FD from another species. Any anti-FD antibody moiety described herein can be used in the anti-human FD antibody construct described herein.

[0142] Factor D, also known as adipsin, is a 24 kDa serine protease containing 228 amino acids. The structure of factor D contains two antiparallel β-barrel domains, each containing six β-chains; all enzymes have the same type of β-chains. Unlike most complement system proteins synthesized by the liver and immune cells, factor D is primarily produced and secreted into the bloodstream by adipocytes. However, macrophages and monocytes can also synthesize this substance, and small amounts are synthesized by astrocytes in the brain. Serum levels of factor D may vary, but under normal conditions, they are found in the range of 1–2 µg / mL. In a healthy state, factor D, like other low-molecular-weight proteins, is filtered by the glomerulus and almost completely reabsorbed in the renal tubules, then rapidly metabolized intracellularly.

[0143] Factor D is produced as a precursor or prozymogen (pre-factor D), requiring subsequent cleavage of a 6-amino acid peptide for maturation. The conversion of pre-factor D to its mature form appears to be rapid, occurring either during or immediately after secretion in the secretory pathway. Despite some controversy, the maturation of pre-factor D to mature factor D is believed to occur primarily through the activation of mannose-binding lectin-associated serine protease-3 (MASP-3), a MASP believed to function in the lectin pathway. Although enzyme-mediated maturation is required, factor D is primarily present in its mature form in resting blood, likely because MASP-3 lacks physiological inhibitors.

[0144] Because mature factor D is the predominant form in resting blood and because it has no known endogenous inhibitors, it must be highly controlled to prevent inappropriate cleavage of endogenous proteins other than its substrate. Therefore, mature factor D is locked in an inactive state through a self-inhibitory loop. This loop determines the enzyme's low reactivity and extremely high specificity for its substrate, factor B. Importantly, factor B can only be cleaved by factor D when it binds to C3b or C3(H2O). After binding and cleaving factor B, factor D is not permanently incorporated into the complex but is recycled through a reversible reaction. Therefore, glomerular filtration by the kidneys plays a crucial role in regulating factor D concentration. Besides host defense against pathogens, the bypass pathway and factor D are also involved in various other physiological processes. For example, C3b-mediated opsonization is known to be responsible for labeling damaged hepatocytes for clearance by phagocytes following acute liver injury. This process provides a scaffold for the development of newly formed cells and helps prevent persistent inflammation. The bypass pathway and factor D have been shown to be essential in this process. However, the activation level of the alternative pathway needs to be kept in good balance, because an overactive complement cascade is known to lead to widespread hepatocellular death, persistent inflammation, and liver damage.

[0145] Recent studies have shown that factor D is also involved in the skin aging process. With age, the extracellular matrix of the dermis is known to degenerate with an increase in the number of senescent cells. In human skin samples, expression levels in older subjects were higher than in younger subjects. Another tissue in which factor D plays a role is adipose tissue. Adipocytes are energy reservoirs and play a crucial role in energy balance. They are responsible not only for lipolysis but also for glucose uptake and triglyceride synthesis. Interestingly, factor D is not the only complement system component produced in adipose tissue. Other factors, including C3 and factor B, are also expressed to some extent in this tissue, and in the absence of pathogens, they have been found to activate the proximal portion of the alternative pathway (i.e., upstream of C5 cleavage). In fact, studies have shown that factor D plays an important role in adipocyte differentiation and lipid accumulation through C3a signaling. In contrast, activation of the terminal or cleaved portion of this pathway has not been observed in adipose tissue because proteins such as C5 are not expressed. As part of their role in energy homeostasis, adipocytes also have endocrine functions. In response to certain stimuli, they secrete regulatory molecules that play a role in the metabolic functions of other tissues. These regulatory molecules include fatty acids and adipokines, such as factor D. Studies have shown that when blood glucose levels are elevated, factor D indirectly induces insulin secretion from pancreatic β-cells by controlling the production of C3a via the alternative pathway. Furthermore, research has found that factor D / C3a signaling can protect pancreatic β-cells by preventing cell dedifferentiation and death. Studies have even indicated that higher levels of circulating factor D are associated with a lower risk of developing diabetes in middle-aged individuals.

[0146] A deficiency in components of the proximal portion of the alternative pathway (such as factor D) leads to an inability to opsonize invading pathogens and insufficient MAC formation. This ultimately limits phagocytosis and lysis of invaders. Therefore, it is not surprising that complete factor D deficiency has been identified as a risk factor for severe bacterial infections. For example, complete factor D deficiency was observed in a Dutch patient with meningitis due to a Ser42Stop mutation in both alleles of the gene. Complete factor D deficiency is associated with a decreased ability to opsonize and phagocytose bacteria, and this has also been observed in other family members. Other case studies have also found that factor D deficiency is associated with Neisseria gonorrhoeae infection and various respiratory infections. Although the underlying factor D mutations are not always the same, they can all lead to protein instability or abnormal protein folding, resulting in a failure to secrete. Complete factor D deficiency requires mutations in both alleles, therefore, its inheritance pattern is autosomal recessive.

[0147] The anti-FD antibody moiety or its functional fragment can specifically bind to FD from any source, such as any organism with a complement system, including but not limited to dogs, cats, pigs, cattle, sheep, goats, horses, rats, rabbits, hamsters, guinea pigs, monkeys, mice, and humans. In some embodiments, the FD is human FD. In some embodiments, the FD is cynomolgus monkey FD (cynomolgus monkey FD).

[0148] In some implementations, factor D is human factor D. In some implementations, the binding of the anti-FD antibody moiety to human factor D is associated with a reduction in the production of C3bBb in the complement activation pathway of intact organisms.

[0149] In some embodiments, the anti-FD antibody portion binds to human FD. In some embodiments, the anti-FD antibody portion binds only to human FD and not to FDs from another species. In some embodiments, the anti-FD antibody portion is cross-species reactive to FDs other than human FD. Exemplary non-human FDs include, but are not limited to, mouse FD, rat FD, rabbit FD, sheep FD, and cynomolgus monkey FD. In some embodiments, the anti-FD antibody portion cross-reacts with cynomolgus monkey FD (cyno FD). In some embodiments, the anti-FD antibody portion does not cross-react with mouse FD.

[0150] In some embodiments, the anti-FD antibody partially binds to the mature FD molecule. In some embodiments, the anti-FD antibody partially binds to the precursor FD. In some embodiments, the anti-FD antibody partially binds to both the precursor FD and the mature FD.

[0151] The anti-FD antibody moiety can be any suitable form known in the art. In some embodiments, the anti-FD antibody moiety is selected from the group consisting of: full-length antibodies, Fab, Fab', F(ab')2, scFv, and combinations thereof. In some embodiments, the anti-FD antibody moiety comprises a full-length antibody, such as a full-length antibody comprising an Fc fragment derived from IgG4 (or consisting substantially of full-length antibodies). In some embodiments, the anti-FD antibody moiety comprises anti-FD scFv (or consists substantially of anti-FD scFv, or consists of anti-FD scFv). In some embodiments, the anti-FD antibody moiety is a mouse antibody, a chimeric antibody, or a humanized antibody. In some embodiments, the anti-FD antibody moiety is monospecific. In some embodiments, the anti-FD antibody moiety is multispecific. In some embodiments, the anti-FD antibody moiety is monovalent. In some embodiments, the anti-FD antibody moiety is multivalent.

[0152] In some embodiments, the anti-human FD antibody moiety has one or more mutations (e.g., insertions, deletions, and / or substitutions) that reduce off-target binding. In some embodiments, the binding of the anti-FD antibody moiety to FD (e.g., human FD) is pH-dependent, wherein the anti-FD antibody moiety binds to FD at a neutral pH (e.g., about pH 7.4; e.g., the pH found in blood) than it binds to FD at an acidic pH (e.g., about pH 5.8; e.g., the pH found in endosomal tissue). In some embodiments, the binding affinity of the pH-dependent anti-FD antibody moiety to FD (e.g., human FD) at about pH 7.4 is at least about 2 times (e.g., at least about 3, 4, 5, 6, 7, 8, 9, 10 times or higher) than the binding affinity of the pH-dependent anti-FD antibody moiety to FD (e.g., human FD) at about pH 5.8.

[0153] In some embodiments, the anti-FD antibody moiety is affinity-matured. In some embodiments, the affinity-matured anti-FD antibody moiety has a binding affinity for FD (e.g., human FD and / or cyno FD) that is at least about 2 times (e.g., at least about 3, 4, 5, 10, 50, 100, 1000 times or more) than that of the parental anti-FD antibody moiety (i.e., unaffected). In some embodiments, the affinity-matured anti-FD antibody moiety has a binding affinity for human FD similar to that of the parental anti-FD antibody moiety (e.g., within about 2 times), but a binding affinity for cynomolgus monkey FD that is at least about 2 times (e.g., at least about 3, 4, 5, 10, 50, 100, 1000 times or more) than that of the parental anti-FD antibody moiety.

[0154] In some implementations, the anti-FD antibody portion is affinity-matured, and the binding of the anti-FD antibody portion to FD (e.g., human FD) is pH-dependent.

[0155] In some implementations, the anti-human FD antibody portion includes a heavy chain variable domain (VH) and a light chain variable domain (VL).

[0156] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: i) a VH comprising H-CDR1, H-CDR2, and H-CDR3, wherein H-CDR1, H-CDR2, and H-CDR3 respectively comprise the amino acid sequences of H-CDR1 (or a variant thereof comprising up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, e.g., conservative substitutions), H-CDR2 (or a variant thereof comprising up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, e.g., conservative substitutions)) and H-CDR3 (or a variant thereof comprising up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, e.g., conservative substitutions)) within a reference VH having the amino acid sequence shown in SEQ ID NO: 7; and ii) VL, comprising L-CDR1, L-CDR2, and L-CDR3, wherein L-CDR1, L-CDR2, and L-CDR3 respectively comprise L-CDR1 (or a variant thereof comprising up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, e.g., conservative substitutions), L-CDR2 (or a variant thereof comprising up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, e.g., conservative substitutions)), and L-CDR3 (or a variant thereof comprising up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, e.g., conservative substitutions)) within a reference VL having the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the CDR positions are based on Kabat numbers. In some embodiments, the affinity of this anti-human FD antibody for FD (e.g., human FD and / or cynomolgus monkey FD) is comparable to that of a reference antibody for FD (e.g., the difference is in the range of about 2-fold), said reference antibody comprising a VH containing the amino acid sequence of SEQ ID NO: 7 and a VL containing the amino acid sequence of SEQ ID NO: 8. In some embodiments, the affinity of this anti-human FD antibody for FD (e.g., human FD and / or cynomolgus monkey FD) is at least about 2-fold (e.g., at least about 3, 4, 5, 10, 50, 100, 1000-fold or higher) than that of the reference antibody for FD, said reference antibody comprising a VH containing the amino acid sequence of SEQ ID NO: 7 and a VL containing the amino acid sequence of SEQ ID NO: 8.

[0157] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising VH and VL, wherein: (i) the VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 1 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); H-CDR2 containing the amino acid sequence of SEQ ID NO: 2 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and (ii) the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 4 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and H-CDR2 containing the amino acid sequence of SEQ ID NO: 2 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and H-CDR2 containing the amino acid sequence of SEQ ID NO: 3 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations L-CDR2 consisting of the amino acid sequence NO: 5 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions); and L-CDR3 consisting of the amino acid sequence SEQ ID NO: 6 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions). In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising VH and VL, wherein: (i) the VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 1 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); H-CDR2 containing the amino acid sequence of SEQ ID NO: 2 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and (ii) the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 4 or a variant thereof containing up to 3 (e.g., 3, 2, or 1) amino acid variations (e.g., insertion, deletion, and / or substitution, e.g., conservative substitution); and H-CDR3 containing the amino acid sequence of SEQ ID NO: 4. L-CDR2 is an amino acid sequence of 5 or a variant thereof containing up to 3 (e.g., 3, 2 or 1) amino acid variations (e.g., insertions, deletions and / or substitutions, e.g., conservative substitutions); and L-CDR3 is an amino acid sequence containing any of SEQ ID NO: 6, 49, 89 and 97.

[0158] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: (i) VH, the VH comprising H-CDR1 containing the amino acid sequence DTYVH (SEQ ID NO: 1); H-CDR2 containing the amino acid sequence RIDP-X1-X2-G-X3-T-X4-F-X5-PRFQA (SEQ ID NO: 9), wherein X1 is A or H, X2 is N, S or Y, X3 is L or H, X4 is T or H, and X5 is D, V, L or H; and H-CDR3 containing the amino acid sequence AMEY (SEQ ID NO: 3); and (ii) VL, the VL comprising L-CDR1 containing the amino acid sequence SA-X6-SDVS-X7-MY (SEQ ID NO: 10), wherein X6 is R, N or S, and X7 is Y, D or V; and H-CDR2 containing the amino acid sequence X8-TSNLAS (SEQ ID NO: 10). L-CDR2 (252), wherein X8 is D or H; and L-CDR3 containing the amino acid sequence QQWSS-X9-PPW-X10 (SEQ ID NO: 11), wherein X9 is Y or H and X10 is T, R, or H. In some embodiments, the CDR positions are based on Kabat numbers.

[0159] In some embodiments (independent of or as a supplement to the above-described CDR sequence), the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 7, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 7; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 8, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 8. In some embodiments, VH comprises the amino acid sequence of SEQ ID NO: 7, or a variant thereof comprising up to about 10 amino acid variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions) in the VH domain, and VL comprises the amino acid sequence of SEQ ID NO: 8, or a variant thereof comprising up to about 10 amino acid variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions) in the VL domain. In some embodiments, the one or more variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions) are present in one or more CDRs. In some embodiments, the one or more variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions) are present in one or more frame regions (FRs). In some embodiments, two or more variations (e.g., insertions, deletions, and / or substitutions, such as conservative substitutions) are present in both the CDRs and FRs. In some embodiments, i) the amino acid residue at position 24 of the VH is A or T; ii) the amino acid residue at position 74 of the VH is K or T; iii) the amino acid residue at position 77 of the VH is N or S; iv) the amino acid residue at position 97 of the VH is A or T; v) the amino acid residue at position 98 of the VH is R or Y; and / or vi) the amino acid residue at position 70 of the VL is F or Y; wherein the numbering is based on the Kabat numbering system. In some embodiments, the amino acid residue positions in the VH are relative to the VH containing the amino acid sequence of SEQ ID NO: 7, and the amino acid residue positions in the VL are relative to the VL containing the amino acid sequence of SEQ ID NO: 8.

[0160] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 1, H-CDR2 containing the amino acid sequence of SEQ ID NO: 2, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 4, L-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 6. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 7, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 8. In some embodiments, an anti-human FD antibody moiety is provided, the anti-human FD antibody moiety comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 18, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 19. In some embodiments, the anti-human FD antibody moiety comprises VH containing the amino acid sequence of SEQ ID NO: 7 and VL containing the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH containing the amino acid sequence of SEQ ID NO: 18 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 18; and VL, said VL containing the amino acid sequence of SEQ ID NO: 19 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 19. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 18 and VL containing the amino acid sequence of SEQ ID NO: 19.

[0161] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 20, H-CDR2 containing the amino acid sequence of SEQ ID NO: 21, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 22; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 23, L-CDR2 containing the amino acid sequence of SEQ ID NO: 24, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 25. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 26, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 27. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 26 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 26; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 27. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 26 and VL containing the amino acid sequence of SEQ ID NO: 27. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2 and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 34, and L-CDR1, L-CDR2 and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 35.In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 34; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 35 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 35. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 34 and VL containing the amino acid sequence of SEQ ID NO: 35. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 216, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 214. In some embodiments, the anti-human FD antibody portion comprises: VH, the VH comprising the amino acid sequence of SEQ ID NO: 216 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 216; and VL, the VL comprising the amino acid sequence of SEQ ID NO: 214 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 214. In some embodiments, the anti-human FD antibody portion includes VH containing the amino acid sequence of SEQ ID NO: 216 and VL containing the amino acid sequence of SEQ ID NO: 214.

[0162] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 36, H-CDR2 containing the amino acid sequence of SEQ ID NO: 37, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 38; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 39, L-CDR2 containing the amino acid sequence of SEQ ID NO: 40, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 41. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 42, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 43. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 42 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 42; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 43 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 43. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 42 and VL containing the amino acid sequence of SEQ ID NO: 43.

[0163] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 44, H-CDR2 containing the amino acid sequence of SEQ ID NO: 45, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 46; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 47, L-CDR2 containing the amino acid sequence of SEQ ID NO: 48, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 49. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 50, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 51. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 50 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 50; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 51 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 51. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 50 and VL containing the amino acid sequence of SEQ ID NO: 51. In some embodiments, the anti-human FD antibody portion binds to human and / or cynomolgus monkey FD in a pH-dependent manner. In some embodiments, the anti-human FD antibody portion binds more strongly to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about twice that at acidic pH.

[0164] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 52, H-CDR2 containing the amino acid sequence of SEQ ID NO: 53, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 54; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 58, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 59. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 58 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 58; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 59 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 59. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 58 and VL containing the amino acid sequence of SEQ ID NO: 59.

[0165] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 63, L-CDR2 containing the amino acid sequence of SEQ ID NO: 64, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 65. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 66, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 67. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 66 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 66; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 67 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 67. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 66 and VL containing the amino acid sequence of SEQ ID NO: 67.

[0166] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 71, L-CDR2 containing the amino acid sequence of SEQ ID NO: 72, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 73. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 74, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 75. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 74 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 74; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 75 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 75. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 74 and VL containing the amino acid sequence of SEQ ID NO: 75.

[0167] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 76, H-CDR2 containing the amino acid sequence of SEQ ID NO: 77, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 78; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 79, L-CDR2 containing the amino acid sequence of SEQ ID NO: 80, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 81. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 82, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 83. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 82 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 82; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 83 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 83. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 82 and VL containing the amino acid sequence of SEQ ID NO: 83.

[0168] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 84, H-CDR2 containing the amino acid sequence of SEQ ID NO: 85, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 86; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 90, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 91. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 90; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 91 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 91. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 90 and VL containing the amino acid sequence of SEQ ID NO: 91.

[0169] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 92, H-CDR2 containing the amino acid sequence of SEQ ID NO: 93, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 94; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 98, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 99. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 98 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 98; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 99 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 99. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 98 and VL containing the amino acid sequence of SEQ ID NO: 99.

[0170] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 100, H-CDR2 containing the amino acid sequence of SEQ ID NO: 101, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 102; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 106, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 107. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 106 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 106; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 107 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 107. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 106 and a VL containing the amino acid sequence of SEQ ID NO: 107.

[0171] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 108, H-CDR2 containing the amino acid sequence of SEQ ID NO: 109, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 110; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 114, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 115. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 114 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 114; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 115. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 114 and a VL containing the amino acid sequence of SEQ ID NO: 115.

[0172] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 116, H-CDR2 containing the amino acid sequence of SEQ ID NO: 117, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 118; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 122, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 123. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 122 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 122; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 123 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 123. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 122 and a VL containing the amino acid sequence of SEQ ID NO: 123.

[0173] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 168, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 169. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 168; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 169. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 168 and VL containing the amino acid sequence of SEQ ID NO: 169.

[0174] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 184, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 185. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 184 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 184; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 185 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 185. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 184 and a VL containing the amino acid sequence of SEQ ID NO: 185.

[0175] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 4, L-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 6. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 186, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 187. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 186; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 187 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 187. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 186 and a VL containing the amino acid sequence of SEQ ID NO: 187.

[0176] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 1, H-CDR2 containing the amino acid sequence of SEQ ID NO: 2, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 289, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 127. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 289 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 289; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 127 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 127. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 289 and VL containing the amino acid sequence of SEQ ID NO: 127.

[0177] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 190, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 191. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 190 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 190; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 191 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 191. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 190 and VL containing the amino acid sequence of SEQ ID NO: 191.

[0178] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 192, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 193. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 192 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 192; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 193 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 193. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 192 and a VL containing the amino acid sequence of SEQ ID NO: 193.

[0179] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 194, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 195. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 194 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 194; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 195 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 195. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 194 and a VL containing the amino acid sequence of SEQ ID NO: 195.

[0180] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 196, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 197. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 196; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 197 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 197. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 196 and VL containing the amino acid sequence of SEQ ID NO: 197.

[0181] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 198, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 199. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 198 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 198; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 199 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 199. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 198 and a VL containing the amino acid sequence of SEQ ID NO: 199.

[0182] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 156, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 157. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 156 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 156; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 157 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 157. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 156 and VL containing the amino acid sequence of SEQ ID NO: 157.

[0183] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 158, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 159. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 158 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 158; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 159 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 159. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 158 and a VL containing the amino acid sequence of SEQ ID NO: 159.

[0184] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 160, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 161. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 160 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 160; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 161 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 161. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 160 and a VL containing the amino acid sequence of SEQ ID NO: 161.

[0185] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 162, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 163. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 162 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 162; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 163 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 163. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 162 and a VL containing the amino acid sequence of SEQ ID NO: 163.

[0186] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 164, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 165. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 164 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 164; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 165 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 165. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 164 and a VL containing the amino acid sequence of SEQ ID NO: 165.

[0187] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 166, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 167. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 166 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 166; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 167 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 167. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 166 and a VL containing the amino acid sequence of SEQ ID NO: 167.

[0188] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 172, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 173. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 172 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 172; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 173 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 173. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 172 and a VL containing the amino acid sequence of SEQ ID NO: 173.

[0189] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 174, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 175. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 174 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 174; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 175 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 175. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 174 and VL containing the amino acid sequence of SEQ ID NO: 175.

[0190] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 176, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 177. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 176 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 176; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 177 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 177. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 176 and a VL containing the amino acid sequence of SEQ ID NO: 177.

[0191] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 178, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 179. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 178 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 178; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 179 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 179. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 178 and VL containing the amino acid sequence of SEQ ID NO: 179.

[0192] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 180, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 181. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 180 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 180; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 181 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 181. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 180 and a VL containing the amino acid sequence of SEQ ID NO: 181.

[0193] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 182, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 183. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 182 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 182; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 183. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 182 and a VL containing the amino acid sequence of SEQ ID NO: 183.

[0194] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 206, H-CDR2 containing the amino acid sequence of SEQ ID NO: 207, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 208; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 209, L-CDR2 containing the amino acid sequence of SEQ ID NO: 210, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 211. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 212, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 213. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 212 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 212; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 213 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 213. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 212 and VL containing the amino acid sequence of SEQ ID NO: 213. In some embodiments, the anti-human FD antibody portion binds to human and / or cynomolgus monkey FD in a pH-dependent manner. In some embodiments, the anti-human FD antibody portion binds more strongly to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about twice that at acidic pH.

[0195] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 234, H-CDR2 containing the amino acid sequence of SEQ ID NO: 235, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 236; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 237, L-CDR2 containing the amino acid sequence of SEQ ID NO: 238, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 239. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 253, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 254. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 253 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 253; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 254 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 254. In some embodiments, the anti-human FD antibody portion comprises VH containing the amino acid sequence of SEQ ID NO: 253 and VL containing the amino acid sequence of SEQ ID NO: 254. In some embodiments, the anti-human FD antibody portion binds to human and / or cynomolgus monkey FD in a pH-dependent manner. In some embodiments, the anti-human FD antibody portion binds more strongly to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about twice that at acidic pH.

[0196] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 240, H-CDR2 containing the amino acid sequence of SEQ ID NO: 241, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 242; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 243, L-CDR2 containing the amino acid sequence of SEQ ID NO: 244, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 245. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 255, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 256. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 255 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 255; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 256 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 256. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 255 and a VL containing the amino acid sequence of SEQ ID NO: 256. In some embodiments, the anti-human FD antibody portion binds to human and / or cynomolgus monkey FD in a pH-dependent manner. In some embodiments, the anti-human FD antibody portion binds more strongly to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about twice that at acidic pH.

[0197] In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising: VH, the VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 246, H-CDR2 containing the amino acid sequence of SEQ ID NO: 247, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 248; and VL, the VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 249, L-CDR2 containing the amino acid sequence of SEQ ID NO: 250, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 251. In some embodiments, an anti-human FD antibody portion is provided, the anti-human FD antibody portion comprising H-CDR1, H-CDR2, and H-CDR3 of VH containing the amino acid sequence of SEQ ID NO: 257, and L-CDR1, L-CDR2, and L-CDR3 of VL containing the amino acid sequence of SEQ ID NO: 258. In some embodiments, the anti-human FD antibody portion comprises: VH, said VH comprising the amino acid sequence of SEQ ID NO: 257 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 257; and VL, said VL comprising the amino acid sequence of SEQ ID NO: 258 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher) amino acid sequence homology with SEQ ID NO: 258. In some embodiments, the anti-human FD antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 257 and a VL containing the amino acid sequence of SEQ ID NO: 258. In some embodiments, the anti-human FD antibody portion binds to human and / or cynomolgus monkey FD in a pH-dependent manner. In some embodiments, the anti-human FD antibody portion binds more strongly to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about twice that at acidic pH.

[0198] In some embodiments, the anti-human FD antibody portion is a full-length antibody (anti-human FD full-length antibody). In some embodiments, the anti-human FD full-length antibody includes an Fc region, such as the human Fc region. In some embodiments, the Fc region is derived from an IgG molecule, such as any of the IgG1, IgG2, IgG3, or IgG4 subclasses. In some embodiments, the anti-human FD full-length antibody includes a heavy chain constant region derived from IgG4 (e.g., human IgG4). In some embodiments, the Fc region is capable of mediating antibody effector functions, such as ADCC and / or CDC. For example, IgG1, IgG2, and IgG3 subclass antibodies with wild-type Fc sequences typically exhibit complement activation, including C1q and C3 binding, while IgG4 does not activate the complement system and does not bind C1q and / or C3. In some embodiments, the Fc region includes modifications that reduce the binding affinity of the Fc region to the Fc receptor (FcR). In some embodiments, the Fc region is derived from IgG4 Fc (e.g., human IgG4 Fc). In some embodiments, the heavy chain constant region includes the amino acid sequence of the wild-type human IgG4 heavy chain constant region. In some implementations, the IgG4 heavy chain constant region or Fc region contains a mutation. See, for example, Armour KL et al. Eur J. Immunol.1999; 29: 2613; and Shields RL et al., J. Biol. Chem. 2001;276: 6591. In some embodiments, the heavy chain constant region comprises one or more mutations (e.g., insertions, deletions, and / or substitutions) in the hinge region for improving stability. In some embodiments, the heavy chain constant region comprises an S228P substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In one embodiment, the IgG4 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 132. In some embodiments, the heavy chain constant region comprises one or more mutations (e.g., insertions, deletions, and / or substitutions) for increasing (e.g., increasing by at least about 20%, 50%, 80%, 90%, 1.5-fold, 2-fold, 5-fold, 10-fold, 20-fold, or higher) binding to neonatal FcR (FcRn; e.g., human and / or cynomolgus monkey FcRn), also referred to herein as "FcRn variants". Any FcRn variant that enhances the binding of the full-length anti-human FD antibody to FcRn (e.g., at about pH 5.8 and / or about pH 7.4) may be used herein. In some embodiments, the heavy chain constant region comprises an M428L / N434A (“LA”) substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In some embodiments, the heavy chain constant region comprises an M428L / N434A (“LA”) substitution (EU number) relative to the heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 133. In some embodiments, the heavy chain constant region comprises an M252Y / V308P / N434Y (“YPY”) substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In some embodiments, the heavy chain constant region contains an M252Y / V308P / N434Y (“YPY”) substitution (EU number) relative to the heavy chain constant region containing the amino acid sequence of SEQ ID NO: 132. In some embodiments, the heavy chain constant region contains an M252Y / N286E / N434Y (“YEY”) substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In some embodiments, the heavy chain constant region contains an M252Y / N286E / N434Y (“YEY”) substitution (EU number) relative to the heavy chain constant region containing the amino acid sequence of SEQ ID NO: 132. In some embodiments, the heavy chain constant region contains the amino acid sequence of SEQ ID NO: 135.In some embodiments, the heavy chain constant region contains an L432C / H433S / N434W / Y436L / T437CE (“N3E”) substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In some embodiments, the heavy chain constant region contains an L432C / H433S / N434W / Y436L / T437CE (“N3E”) substitution (EU number) relative to the heavy chain constant region containing the amino acid sequence of SEQ ID NO: 132. “T437CE” indicates the amino acid substitution T437C followed by an insertion of E immediately after position 437 (EU number). In some embodiments, the heavy chain constant region contains the amino acid sequence of SEQ ID NO: 136. In some embodiments, the heavy chain constant region contains an L432C / H433S / N434W / Y436L / T437C (“N3”) substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In some embodiments, the heavy chain constant region contains a substitution of L432C / H433S / N434W / Y436L / T437C (“N3”) relative to the heavy chain constant region containing the amino acid sequence of SEQ ID NO: 132 (EU number). In some embodiments, the heavy chain constant region contains the amino acid sequence of SEQ ID NO: 137. In some embodiments, the heavy chain constant region contains a substitution of M252Y / S254T / T256E (“YTE”) relative to the wild-type human IgG4 heavy chain constant region sequence (EU number). In some embodiments, the heavy chain constant region contains a substitution of M252Y / S254T / T256E (“YTE”) relative to the heavy chain constant region containing the amino acid sequence of SEQ ID NO: 132 (EU number). In some embodiments, the heavy chain constant region contains the amino acid sequence of SEQ ID NO: 138. In some embodiments, the heavy chain constant region contains an M252Y / S254T / T256E / L432E / H433R / N434F / Y436R / T437Q (“Y31-YTE”) substitution (EU number) relative to the wild-type human IgG4 heavy chain constant region sequence. In some embodiments, the heavy chain constant region contains an M252Y / S254T / T256E / L432E / H433R / N434F / Y436R / T437Q (“Y31-YTE”) substitution (EU number) relative to the heavy chain constant region containing the amino acid sequence of SEQ ID NO: 132. In some embodiments, the heavy chain constant region contains the amino acid sequence of SEQ ID NO: 139. In some embodiments, any mutations (e.g., insertions, deletions, and / or substitutions) described herein that increase the binding of Fc to FcRn may be combined.For example, the “N3E” mutation can be combined with the “YTE” mutation to produce the “N3E-YTE” FcRn variant. In some embodiments, the S228P mutation (EU number) can be combined with any FcRn mutation described herein. In some embodiments, the heavy chain constant region of the anti-human FD full-length antibody (e.g., any anti-human FD full-length antibody described herein) contains the amino acid sequence of any of SEQ ID NO: 132-139.

[0199] In some embodiments, the anti-human FD antibody portion is a full-length anti-human FD antibody, said antibody: i) binds to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) more strongly than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about 2 times (e.g., at least about 2.5 times, 3 times, 3.5 times, 5 times, 10 times or more) than the binding affinity at acidic pH; and ii) binds to FcRn at about pH 5.8 and / or about pH 7.4. The binding affinity of (e.g., human FcRn and / or cynomolgus monkey FcRn) to FD is stronger than that of a reference antibody without FcRn mutation to FD. For example, the binding affinity of a full-length anti-human FD antibody to FcRn is at least about 2 times (e.g., at least about 5, 10, 50, 100, 1000 or higher) than that of a reference antibody without FcRn mutation to FcRn. In some embodiments, the anti-human FD antibody portion is a full-length anti-human FD antibody, said antibody: i) binds to FD (e.g., human FD and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) more strongly than it binds to FD at acidic pH (e.g., about pH 5.8), for example, the binding affinity at neutral pH is at least about 2 times (e.g., at least about 2.5, 3, 3.5, 5, 10 times or more) the binding affinity at acidic pH; and ii) is affinity mature, for example, the binding affinity to FD (e.g., human FD and / or cynomolgus monkey FD) is at least about 2 times (e.g., at least about 3, 5, 10, 20 times or more) the binding affinity of its unaffected parent antibody to FD. In some embodiments, the anti-human FD antibody portion is a full-length anti-human FD antibody, said antibody: i) binds to FcRn (e.g., human FcRn and / or cynomolgus monkey FcRn) more strongly at about pH 5.8 and / or about pH 7.4 than a reference antibody without FcRn mutation binds to FD, for example, the binding affinity of said full-length anti-human FD antibody to FcRn is at least about 2 times (e.g., at least about 5, 10, 50, 100, 1000 times or more) than the binding affinity of the reference antibody without FcRn mutation to FcRn; and ii) is affinity mature, for example, the binding affinity to FD (e.g., human FD and / or cynomolgus monkey FD) is at least about 2 times (e.g., at least about 2.5, 3, 3.5, 5, 10 times or more) than the binding affinity of its unaffected parent antibody to FD.

[0200] In some embodiments, the anti-human FD antibody portion is scFv (anti-human FD scFv). In some embodiments, the anti-human FD scFv comprises from N' to C': VH-optional linker-VL. In some embodiments, the anti-human FD scFv comprises from N' to C': VL-optional linker-VH. Any suitable linker (see, for example, the “Linker” section below) may be used herein, including but not limited to any of SEQ ID NO: 221-229. In some embodiments, an anti-human FD scFv comprising the amino acid sequence of SEQ ID NO: 219 or 220 is provided.

[0201] In addition, anti-human FD antibody moieties and anti-human FD antibody constructs are provided that compete with any of the anti-human FD antibody moieties or anti-human FD antibody constructs described herein for binding to FD (e.g., human FD and / or cynomolgus monkey FD).

[0202] Fc structural domain

[0203] In some embodiments, the isolated anti-human FD antibody construct or anti-human FD antibody moiety includes an Fc domain. In some embodiments, the anti-human FD antibody moiety is a full-length anti-human FD antibody.

[0204] In some embodiments, the Fc domain is an Fc effector domain, that is, an Fc domain that has some or all of the effector functions (including, for example, complement and ADCC functions). In some embodiments, the Fc effector domain is the Fc region of IgG1 or IgG3.

[0205] In some implementations, one or more amino acid modifications may be introduced into the Fc domain of the antibody moiety, thereby creating an Fc domain variant. The Fc domain variant may contain a human Fc domain sequence (e.g., human IgG1, IgG2, IgG3, or IgG4 Fc region) with amino acid modifications (e.g., substitutions) at one or more amino acid positions.

[0206] In some implementations, the Fc domain has some, but not all, effector functions that make the Fc domain a desirable candidate for applications where the in vivo half-life of the antibody moiety is important, but certain effector functions (such as complement and ADCC) are unnecessary or detrimental.

[0207] In vitro and / or in vivo cytotoxicity assays can be performed to analyze the CDC and / or ADCC activity of the Fc domain. For example, an Fc receptor (FcR) binding assay can be performed to determine whether the antibody has FcγR binding (and therefore may have ADCC activity), and / or retains FcRn binding ability. NK cells, the main cells mediating ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Ravetch and Kinet. Annu. Rev. Immunol. Table 2 on page 464 of 9:457-492 (1991). Non-limiting examples of in vitro assays for evaluating the ADCC activity of molecules of interest are described in U.S. Patent No. 5,500,362 (see, for example, Hellstrom, I. et al.). Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)); and Hellstrom, I et al. Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 (See Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assays may be used (see, for example, the ACTI™ non-radioactive cytotoxicity assay for flow cytometry (Cell Technology, Inc. Mountain View, CA); and CytoTox 96). ® Non-radioactive cytotoxicity assays (Promega, Madison, WI). Effector cells that can be used for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or additionally, ADCC activity of the molecule of interest can be assessed in vivo, such as in animal models, e.g., as disclosed in Clynes et al., Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). C1q binding assays can also be performed to confirm that the antibody cannot bind C1q and therefore lacks CDC activity. See, for example, C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, CDC assays can be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood103:2738-2743 (2004)). FcRn binding and in vivo clearance / half-life determination can also be performed using methods known in the art (see, for example, Petkova, SB, et al.). , Int'l. Immunol. 18(12):1759-1769 (2006)).

[0208] Antibodies with reduced effector function include those with substitutions at one or more of the Fc domain residues 238, 265, 269, 270, 297, 327, and 329 (US Patent No. 6,737,056). These Fc mutants include Fc mutants with substitutions at two or more of the amino acid positions 265, 269, 270, 297, and 327, including the so-called “DANA” Fc mutant with residues 265 and 297 substituted for alanine (US Patent No. 7,332,581). In some embodiments, the Fc domain of the multispecific construct or isolated anti-C5 antibody construct does not contain mutations that reduce its effector function, such as one or more mutations described herein. In some embodiments, the Fc domain of the multispecific construct or isolated anti-C5 antibody construct contains one or more of these mutations.

[0209] Certain antibody variants that improve or reduce binding to FcR have been described. See, for example, U.S. Patent No. 6,737,056; WO 2004 / 056312; and Shields et al. , J. Biol. Chem. 9(2): 6591-6604 (2001).

[0210] In some embodiments, the Fc domain is an IgG1 Fc domain. In some embodiments, the IgG1 Fc domain does not contain the L234A mutation and / or the L235A mutation. In some embodiments, the IgG1 Fc domain contains the L234A mutation and / or the L235A mutation (“LALA” mutation). In some embodiments, the Fc domain is an IgG3 Fc domain. In some embodiments, the Fc domain is an IgG2 or IgG4 Fc domain. In some embodiments, the Fc domain is a human IgG4 Fc containing the amino acid sequence of SEQ ID NO: 231. In some embodiments, the Fc domain is an IgG4 Fc domain containing the S228P, L234A, and / or L235A mutation. In some embodiments, the Fc domain is an IgG4 Fc fragment containing the S228P mutation, for example, containing the amino acid sequence of SEQ ID NO: 232. In some implementations, the Fc domain is an IgG4 Fc fragment containing the S228P / M428L / N434A (“PLA”) triple mutation, such as the amino acid sequence containing SEQ ID NO: 230.

[0211] In some embodiments, the anti-human FD antibody moiety (or isolated anti-human FD antibody construct) includes an Fc domain with one or more amino acid substitutions that improve ADCC, for example, substitutions at positions 298, 333 and / or 334 (EU numbers of residues) in the Fc domain.

[0212] In some implementations, the Fc domain is modified to alter (i.e., improve or reduce) C1q binding and / or complement-dependent cytotoxicity (CDC), as described in U.S. Patent Nos. 6,194,551, WO 99 / 51642, and by Idusogie et al. J. Immunol. As described in 164: 4178-4184 (2000).

[0213] In some embodiments, the anti-human FD antibody moiety (or isolated anti-human FD antibody construct) includes a variant Fc domain containing one or more amino acid substitutions that alter the half-life and / or change the binding to FcRn. Antibodies with increased half-life and improved binding to FcRn (responsible for transferring maternal IgG to the fetus) (Guyer et al.) J. Immunol. 117:587 (1976) and Kim et al., J. Immunol.24:249 (1994) is described in US2005 / 0014934A1 (Hinton et al.). These antibodies contain an Fc domain having one or more substitutions that alter the binding of the Fc domain to FcRn. Such Fc variants include those that have substitutions at one or more Fc domain residues (e.g., substitution at Fc domain residue 434 (US Patent No. 7,371,826)).

[0214] For other examples of variants of the Fc domain, see also Duncan and Winter. Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821; and WO 94 / 29351.

[0215] Binding affinity of anti-human FD antibody moiety

[0216] The binding affinity and specificity of the anti-human FD antibody moiety described herein can be experimentally determined using methods known in the art. For example, antibody-protein antigen binding can be detected and / or quantified using a variety of techniques, including but not limited to Western blot, dot blot, surface plasmon resonance (SPR) methods (e.g., BIAcore system; Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, NJ), biomembrane interferometry (BLI) (e.g., Octet system, ForteBio), RIA, ECL, IRMA, EIA, peptide scanning, and enzyme-linked immunosorbent assay (ELISA). Furthermore, methods for measuring affinity (e.g., measuring dissociation and association constants via BLI) are also described in the working examples.

[0217] In some embodiments, the anti-human FD antibody partially and specifically binds to FD (e.g., human FD and / or cynomolgus monkey FD), where K D It is about 10 -7 M to approximately 10 -12 M, for example, any of the following: approximately 10 -7 M to approximately 10 -10 M, approximately 10 -8 M to approximately 10 -11 M, approximately 10 -9 M to approximately 10 -11 M, approximately 10 -8 M to approximately 10 -10 M, approximately 10 -8 M to approximately 10 -9 M, approximately 10-9 M to approximately 10 -10 M, or about 10 -10 M to approximately 10 -11 M. In some embodiments, the anti-human FD antibody partially and specifically binds to FD (e.g., human FD and / or cynomolgus monkey FD), wherein K D It is about 10 -8 M to approximately 10 -12 M, or approximately 1×10 -9 M to approximately 1×10 -12 M.

[0218] In some embodiments, the anti-human FD antibody moiety binds more strongly to human FD and / or cynomolgus monkey FD at neutral pH (e.g., about pH 7.4) than it does at acidic pH (e.g., about pH 5.8). In some embodiments, the anti-human FD antibody moiety specifically binds to FD (e.g., human FD and / or cynomolgus monkey FD), wherein at about pH 5.8, K D It is approximately 5 × 10 -10 M to approximately 10 × 10 -10 M. In some embodiments, the anti-human FD antibody partially and specifically binds to FD (e.g., human FD and / or cynomolgus monkey FD), wherein at approximately pH 7.4, K D It is approximately 1×10 -10 M to approximately 5 × 10 -10 M.

[0219] Biological characteristics of anti-human FD antibody

[0220] Because C3 spontaneously hydrolyzes to form C3(H2O), AP is considered to have constitutive activity at low levels. C3(H2O) behaves similarly to C3b because it can associate with fB, making fB readily cleaved and activated by fD. The resulting C3(H2O)Bb then cleaves C3 to produce C3b and C3a, initiating the AP cascade by forming the C3 convertase C3bBb of AP. As the initial C3 convertase produces increasing amounts of C3b, an amplification loop is established. It should be noted that since CP and LP also produce C3b, which can bind to factor B and conjugate AP, the AP amplification loop also participates in CP and LP once these pathways are activated. Therefore, AP consists of two functional entities: an independent complement activation pathway independent of CP or LP, and an amplification process that participates in and facilitates the full expression of CP and LP.

[0221] In some embodiments, the anti-human FD antibody partially inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) AP. In some embodiments, the anti-human FD antibody partially inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) AP amplification process or amplification loop. In some embodiments, the anti-human FD antibody partially does not inhibit (or inhibits at most about 10%, 5%, 2%, 1%, or less) CP or LP activation. In some embodiments, the anti-human FD antibody portion or its use preserves the individual's ability to fight infection through CP and LP. In some embodiments, the anti-human FD antibody partially inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) amplification process or amplification loop. In some implementations, the anti-FD antibody partially inhibits the IC50 of CP and / or LP (e.g., inhibiting the lysis of sheep RBCs in 20% human serum). 50 The value could not be detected.

[0222] In some embodiments, the AP activity partially inhibited by the method of the present invention or by the anti-human FD antibody described herein is AP activation induced by one or more of the following: lipopolysaccharide (LPS), lipooligosaccharide (LOS), pathogen-associated molecular patterns (PAMPs), and danger-associated molecular patterns (DAMPs). In some embodiments, the AP activity partially inhibited by the method of the present invention or by the anti-human FD antibody described herein is the production of the C3bBb protein complex. In some embodiments, the AP activity partially inhibited by the method of the present invention or by the anti-human FD antibody described herein is FD-dependent.

[0223] Methods for determining whether a particular antibody described herein inhibits human FD are known in the art. Inhibition of human FD reduces APC-mediated cytolysis in the body fluids of a subject. This reduction in cytolysis present in the body fluids can be measured by methods well known in the art, such as by conventional hemolysis assays, such as hemolysis assays in chicken erythrocyte hemolysis methods, as described in Hillmen et al. (2004) N Engl. J Med 350(6):552; and Yuan et al. (2017) Haematologica 102(3):466-475. The concentration and / or physiological activity of FD in the body fluids can be measured by methods well known in the art. Methods for measuring FD concentration or activity include, for example, ELISA (see, for example, Corvillo et al. (2021) Int J Mol Sci 22(12):6608). Inhibition of FD can cause inhibition of alternative pathway activity (e.g., C3b production), as illustrated in Barratt et al. (2021) Front Immunl. 12: 712572. Therefore, for example, ELISA assays can be used to determine the inhibitory effect of anti-FD antibodies on LPS-induced C3b deposition, as described, for example, in Kimura et al. (2008) Blood. 111(2):732–740; Li et al. (2020) Front Immunol. 11:1123; and “Instructions for Use WIESLAB® Complement System Alternative Pathway”, document number LABEL-DOC-0034, 2.0, December 2018. Other assays known in the art can also be used. Candidates capable of inhibiting human FD can be screened using these or other suitable types of assays.

[0224] Hemolysis assays can be used to determine the inhibitory activity of anti-FD antibodies against FD-mediated complement activation, and thereby to determine the potential off-target binding of anti-FD antibodies. These assays include, but are not limited to, sheep erythrocyte (RBC) lysis assays. To determine the effect of humanized anti-FD antibodies on complement pathway-mediated hemolysis in in vitro serum test solutions, for example, rabbit RBC lysis assays can be used to examine AP regulation, or LPS-based ELISA assays can be used to examine AP complement inhibition. The percentage of lysis is normalized by considering 100% lysis as lysis occurring in the absence of inhibitors. C3 inhibition can be measured by methods known in the art, including, for example, flow cytometry and commercial kits (e.g., Wieslab® assays). See, for example, Mannes et al., (2021). Blood137(4):443-455; and “Instructions for UseWIESLAB® Complement System Alternative Pathway”, document number LABEL-DOC-0034, 2.0, December 2018.

[0225] In some embodiments, the binding of the anti-human FD antibody portion to FD is associated with a reduction in the production of C3bBb in the complement activation pathway of an intact organism (e.g., human) (e.g., a reduction of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%). In some embodiments, the anti-human FD antibody portion inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) LPS-induced C3b and / or C5b-9 deposition. In some embodiments, the anti-FD antibody portion does not inhibit (or inhibits at most about 10%, 5%, 2%, 1%, or less) IgM-induced or mannan-induced C3b and / or C5b-9 deposition.

[0226] Methods for testing the activity of the anti-human FD antibody moiety (e.g., AP inhibition) are well known in the art, including but not limited to rabbit RBC lysis assays (e.g., induced by human serum) or LPS-induced C3b and / or C5b-9 deposition assays. See, for example, the assay in US11434279, the contents of which are incorporated herein by reference in their entirety. See also Example 4 herein.

[0227] pH-dependent and / or FcRn binding optimized anti-human FD antibody moiety

[0228] In some embodiments, the anti-human FD antibody portion described herein exhibits pH-dependent dissociation from factor D (e.g., human FD and / or cynomolgus monkey FD). This pH-dependent binding results in greater persistence of the administered antibody or antibody fusion protein molecule because the immune complex taken up by the cell (i.e., the anti-human FD antibody construct bound to factor D) will dissociate in the acidic environment of the endosome, allowing the released antibody or antibody fusion protein to recycle out of the cell via the neonatal Fc receptor (FcRn), where it can bind to new factor D molecules.

[0229] As used herein, the term "pH-dependent binding" means that the antibody exhibits reduced binding to FD at acidic pH (e.g., about pH 5.8; e.g., in the body) compared to its binding to FD at neutral pH (e.g., about pH 7.4; e.g., in the blood) (e.g., a reduction of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%).

[0230] The pH dependence of the anti-human FD antibody moiety described herein can be experimentally determined by methods known in the art, such as those described in US20220204602, US20220177556, US9,079,949, and US9765135, the contents of which are incorporated herein by reference in their entirety. See also Example 3 herein. The pH dependence of FD binding can reflect differences in binding properties, such as binding affinity (e.g., dissociation constant), kinetic parameters (e.g., association rate and dissociation rate), and dissociation percentage at different pH values. In some embodiments, the pH dependence of the anti-human FD antibody moiety can be expressed as a ratio of dissociation percentages. In some embodiments, the dissociation percentage can be expressed as a low-pH dissociation factor and a neutral-pH dissociation factor. In some embodiments, the pH dependence of the anti-human FD antibody moiety is expressed as a Kc of approximately pH 5.8 versus approximately pH 7.4. off The ratio is expressed. In some embodiments, the pH dependence of the anti-human FD antibody moiety is approximately pH 5.8 versus approximately pH 7.4 with K. D Ratio representation.

[0231] The pH dependence of the binding of anti-human FD antibody to FD can be assessed based on the dissociation of FD-bound antibody at acidic pH (e.g., approximately pH 5.8) or neutral pH (e.g., approximately pH 7.4). The dissociation of FD-bound antibody at acidic pH can be determined using a low-pH dissociation factor, which is the percentage of antibody dissociated from the antigen at approximately pH 5.8 and a temperature of approximately 25°C (where the antibody is pre-bound to the antigen at approximately pH 7.4). The low-pH dissociation factor can be measured by associating the antibody with the antigen (e.g., an anti-human FD antibody construct and human / cynomolgus monkey factor D) at pH 7.4 for approximately 600 seconds, followed by dissociation in a buffer at approximately pH 5.8 for approximately 600 seconds, and calculating the percentage of antibody dissociated from the antigen at approximately pH 5.8. In some embodiments, the low pH dissociation factor of the anti-human FD antibody moiety (i.e., the percentage of the anti-human FD antibody moiety that dissociates from bound FD at an acidic pH (e.g., about pH 5.8) is not less than any one of about 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%).

[0232] The dissociation factor of FD-binding antibodies at neutral pH can be used, which is the percentage of antibody dissociation from antigen at approximately pH 7.4 and a temperature of approximately 25°C (where the antibody is pre-bound to the antigen at approximately pH 7.4), to determine the dissociation of FD-binding antibodies at neutral pH. The neutral pH dissociation factor can be measured by associating the antibody with the antigen (e.g., an anti-human FD antibody construct and human / cynomolgus monkey FD) at approximately pH 7.4 for approximately 600 seconds, followed by dissociation in a buffer solution at approximately pH 7.4 for approximately 600 seconds, and calculating the percentage of antibody dissociated from the antigen at approximately pH 7.4. In some embodiments, the neutral pH dissociation factor of the anti-human FD antibody portion (i.e., the percentage of the anti-human FD antibody portion dissociated from bound FD at neutral pH (e.g., approximately pH 7.4) does not exceed any one of approximately 20%, 18%, 16%, 14%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0233] In some embodiments, the ratio of the low-pH dissociation factor to the neutral-pH dissociation factor of the anti-human FD antibody portion of the present invention is at least about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 20, 50 or higher. In some embodiments, the ratio of the percentage of antibody dissociation of factor D at pH 5.8 to the percentage of antibody dissociation of factor D at pH 7.4 is at least about 3, 4, 5 or 6.

[0234] In some embodiments, the anti-human FD antibody portion binds more strongly to FD (e.g., human and / or cynomolgus monkey FD) at neutral pH (e.g., about pH 7.4) than it binds to FD at acidic pH (e.g., about pH 5.8) (e.g., at least about 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5 times or more).

[0235] In some embodiments, the FcRn variant anti-human FD antibody portion described herein exhibits a prolonged in vivo serum half-life (e.g., an increase of at least about 10%, 20%, 50%, 70%, 90%, 1.5-fold, 2-fold, 5-fold, 20-fold, or higher) compared to a reference anti-human FD antibody without an FcRn binding mutation, for example in mice (including transgenic mice). In some embodiments, the anti-human FD antibody portion exhibits a prolonged serum half-life in other test animals, including but not limited to rats, chickens, rabbits, sheep, and cynomolgus monkeys. In some embodiments, the anti-human FD antibody portion exhibits a prolonged serum half-life in humans.

[0236] Antibodies with pH-dependent antigen binding can enhance the degradation of antigens in endosomes or lysosomes; and more free or unbound antibodies are recovered via FcRn binding and eventually recycled back into the circulation to bind to additional antigens (Igawa et al., Nat Biotechnol. Nov 2010; 28(11):1203-7). pH-dependent antigen binding properties can arise naturally or can be generated by engineering antigen-binding interactions.

[0237] IgG Fc naturally binds to FcRn in a pH-dependent manner. Engineered antibodies possess enhanced pH-dependent FcRn binding properties, such as increased binding to FcRn at acidic pH (e.g., about pH 5.8) (e.g., an increase of at least about 2-fold, 5-fold, 20-fold, or higher) and minimal (e.g., less than 2-fold) or no change in binding at neutral pH (e.g., about pH 7.4). These antibodies can be more effectively recovered and recycled back to serum and are also known as "recirculating antibodies." For example, increasing the affinity of antibody Fc for FcRn at acidic pH in the body can improve antibody serum persistence and has been used in the development of therapeutic antibodies. Recirculating antibodies can have an extended serum half-life. Engineered antibodies Fc that exhibit enhanced binding affinity to FcRn at acidic pH (e.g., approximately pH 5.8) and neutral pH (e.g., approximately pH 7.4) in the endosome can enhance antigen / antibody complexes that are taken up from the circulation (to reduce serum antigen levels) and recirculated from the endosome back into the circulation to bind more neoantigens; these are therefore termed "scavenging" antibodies (Igawa et al., PloS One. May 7, 2013; 8(5):e63236). For example, FcRn binding mutant LA has been shown to increase antibody recycling, thereby prolonging serum IgG persistence (Yeung et al., ...). J Immunol (2009) 182 (12): 7663–7671). FcRn binding mutations Y31-YTE and N3E (Borrok et al., J Biol Chem February 13, 2015; 290(7):4282-90) and YEY and YPY (Igawa et al., PloS One. May 7, 2013; 8(5):e63236) have been shown to significantly increase FcRn binding at both pH 7.4 and pH 5.8. Antibodies carrying these Fc mutations have been observed to persist in serum for an extended period and / or have decreased serum antigen levels. The contents of these references are incorporated herein by reference in their entirety.

[0238] In some embodiments, the binding of the anti-human FD antibody moiety to FD (e.g., human and / or cynomolgus monkey FD) is pH-dependent, and dissociation from FD at acidic pH (e.g., about pH 5.8) is faster than at neutral pH (e.g., about pH 7.4) by at least about 50%, 70%, 90%, 1.5-fold, 2-fold, 5-fold, 20-fold, or higher. In some embodiments, the binding of the anti-human FD antibody moiety (e.g., full-length anti-human FD antibody) to FcRn is improved (e.g., increased by at least about 10%, 20%, 50%, 70%, 90%, 1.5-fold, 2-fold, 5-fold, 20-fold, 100-fold, 1000-fold, or higher) at about pH 7.4 and / or about pH 5.8, for example, compared to a reference anti-human FD antibody without an FcRn binding mutation. In some embodiments, the anti-human FD antibody moiety is a recycled antibody moiety. In some embodiments, the anti-human FD antibody portion is a scavenging antibody portion. In some embodiments, the anti-human FD antibody portion is a recycling antibody portion and a pH-dependent FD-binding antibody portion. In some embodiments, the anti-human FD antibody portion is a scavenging antibody portion and a pH-dependent FD-binding antibody portion. In some embodiments, the full-length anti-human FD antibody has a binding affinity of about 10 for FcRn at about pH 7.4. -11 M to approximately 10 -5 M, for example, about 10 -10 M to approximately 10 -6 M, or about 10 -9 M to approximately 10 -8 M. In some implementations, the binding affinity of the full-length anti-human FD antibody to FcRn is approximately 10 at approximately pH 5.8. -11 M to approximately 10 -7 M, for example, about 10 -11 M to approximately 10 -8 M, or about 10 -10 M to approximately 10 -8 M.

[0239] Fusion coordinators and multispecific constructs

[0240] In some embodiments, the isolated anti-human FD antibody construct described herein further comprises a fusion coordinate, or a second antibody portion that specifically recognizes a complement pathway component (hereinafter also referred to as a "complement protein"). Therefore, in some embodiments, an isolated anti-human FD antibody construct is provided, comprising: an anti-human FD antibody portion (e.g., any anti-human FD antibody portion described herein) and a second antibody portion that specifically recognizes a complement pathway component (e.g., FD or non-FD complement protein). Any complement protein may be used herein. Exemplary fusion coordinates include, but are not limited to, an anti-FD antibody portion (e.g., any anti-human FD antibody portion described herein), an anti-C2 antibody portion, or an anti-C5 antibody portion. In some embodiments, the isolated anti-human FD antibody construct is monospecific. In some embodiments, the isolated anti-human FD antibody construct is multivalent. In some embodiments, the isolated anti-human FD antibody construct is multispecific ("multispecific construct").

[0241] In some embodiments, the second antibody portion is selected from the group consisting of: full-length antibodies, Fab, Fab', F(ab)2, F(ab')2, scFv, sdAb, and combinations thereof. In some embodiments, the second antibody portion is scFv.

[0242] The complement system plays a crucial role in the pathogenesis of many autoimmune, inflammatory, and ischemic diseases. Improper complement activation and complement deposition on host cells can lead to complement-mediated cell and target tissue lysis and / or damage, as well as tissue destruction due to the production of potent inflammatory mediators. The complement system, also known as the complement cascade, is part of the immune system. It enhances the ability of (complementary) antibodies and phagocytes to clear microorganisms and damaged cells from the body, promote inflammation, and attack pathogen cell membranes. It is part of the innate immune system, lacks adaptive response, and does not change throughout an individual's life. However, the complement system can be recruited and function through antibodies produced by the adaptive immune system.

[0243] Unbound by any theory or assumption, three complement pathways are known: the alternative complement pathway (AP), the classical pathway (CP), and the lectin pathway (LP). Generally, CP is initiated by antigen-antibody complexes, LP is activated by lectin binding to sugar molecules on microbial surfaces, and AP, while constitutively active at low levels, can rapidly proliferate on bacterial, viral, and parasitic cell surfaces due to a lack of regulatory proteins. Host cells are usually protected from AP complement activation by regulatory proteins. However, in certain situations, such as when regulatory proteins are defective or absent, AP can also be uncontrolled activated on host cells, leading to complement-mediated diseases or symptoms. CP consists of components C1, C2, and C4, and merges with AP in the C3 activation step. LP consists of mannose-binding lectin (MBL) and MBL-associated serine protease (MASP), and shares components C4 and C2 with CP. AP consists of component C3 and several factors, such as factor B, factor D, propertin, and fluid phase regulator H. Complement activation consists of three phases: (a) recognition, (b) enzymatic activation, and (c) membrane attack leading to cell death. The first phase of CP complement activation begins at C1. C1 is composed of three distinct proteins: the recognition subunit C1q and the serine protease subfractions C1r and C1s, which bind together to form the calcium-dependent tetrameric complex C1r2s2. The intact C1 complex is essential for the physiological activation of C1. Activation occurs when the intact C1 complex binds to an immunoglobulin complex conjugated to an antigen. This binding activates C1s, which then cleaves C4 and C2 proteins to produce C4a and C4b, as well as C2a and C2b. The C4b and C2a fragments combine to form the C3 convertase C4b2a, which in turn cleaves C3 to form C3a and C3b. LP activation is initiated by the binding of MBL to certain sugars on the target surface, which triggers the activation of MBL-associated serine proteases (MASP). MASP then cleaves C4 and C2 in a manner similar to the activity of C1s in CP, resulting in the C3 convertase C4b2a. Therefore, CP and LP are activated through different mechanisms, but they share the same components C4 and C2, and both pathways lead to the production of the same C3 convertase C4b2a. The cleavage of C3 into C3b and C3a by C4b2a is a central event in the complement pathway for two reasons. It initiates the AP amplification loop because surface-deposited C3b is a key intermediate for the AP C3 convertase C3bBb. Both C3a and C3b have important biological significance. C3a has pro-inflammatory effects and, along with C5a, is known as an allergen. C3b and its further cleavage products also bind to complement receptors present on neutrophils, eosinophils, monocytes, and macrophages, thereby promoting the phagocytosis and clearance of C3b opsonization granules.Finally, C3b can associate with C4b2a or C3bBb to form C5 convertases of CP and LP and AP, respectively, activating the terminal complement sequence, leading to the production of the potent pro-inflammatory mediator C5a and the assembly of the soluble membrane attack complex (MAC) C5-C9.

[0244] Complement dysfunction is a cause of several human glomerular diseases, including atypical hemolytic uremic syndrome (aHUS), antineutrophil cytoplasmic antibody-mediated vasculitis (ANCA), C3 glomerulonephropathy, IgA nephropathy, immune complex-induced membranoproliferative glomerulonephritis, renal ischemia-reperfusion injury, lupus nephritis, membranous nephropathy, and chronic transplant-mediated glomerulonephropathy. Abnormal complement component activation has been proposed as a biomarker for various types of cancer and their clinical outcomes. Lung cancer patients show significantly higher plasma complement protein and activation fragment levels than control donors, and elevated complement levels correlate with lung tumor size. Complement-related proteins are also elevated in biofluids from patients with other types of tumors. See, for example, Pio et al., Semin Immunol. 2013 Feb; 25(1): 54–64. Inhibition of the complement cascade has been proposed for the treatment of glomerular diseases and cancer.

[0245] In some embodiments, the second antibody moiety specifically recognizes FD. In some embodiments, the isolated anti-human FD antibody construct comprises two or more anti-human FD antibody moieties (e.g., scFv), such as two or more anti-human FD antibody moieties described herein. In some embodiments, the two or more anti-human FD antibody moieties are fused together in tandem. In some embodiments, the two or more anti-human FD antibody moieties are on different polypeptide chains within the isolated anti-human FD antibody construct. In some embodiments, the two or more anti-human FD antibody moieties bind to the same FD epitope. In some embodiments, the two or more anti-human FD antibody moieties bind to at least two different FD epitopes. In some embodiments, the two or more anti-human FD antibody moieties each bind to a different FD epitope.

[0246] In some embodiments, the second antibody moiety specifically recognizes complement component 2 (“C2”). In some embodiments, the second antibody moiety specifically recognizes complement component 5 (“C5”). In some embodiments, the isolated anti-human FD antibody construct comprises an anti-human FD antibody moiety (e.g., any anti-human FD antibody moiety described herein) and one or more antibody moieties (e.g., scFv) that specifically recognize non-FD complement proteins, said one or more antibody moieties being fused together in tandem with each other or with the anti-human FD antibody moiety, or on separate polypeptide chains.

[0247] In some embodiments, the isolated anti-human FD antibody construct comprises one polypeptide chain (or consists essentially of one polypeptide chain, or consists of one polypeptide chain). In some embodiments, the isolated anti-human FD antibody construct comprises two or more (e.g., 2) polypeptide chains (or consists essentially of two or more polypeptide chains, or consists of two or more polypeptide chains), such as heterodimers or homodimers formed in the presence of an Fc domain.

[0248] In some embodiments, the isolated anti-human FD antibody construct comprises an anti-human FD scFv (“scFv1”; for example, any anti-human FD scFv described herein) fused to a second scFv (scFv2) that specifically recognizes a component of the complement pathway. In some embodiments, scFv2 is an anti-C2 scFv. In some embodiments, scFv2 is an anti-C5 scFv. In some embodiments, the scFv (e.g., scFv1 or scFv2) comprises, from the N-terminus to the C-terminus: VH-optional linker-VL. In some embodiments, the scFv (e.g., scFv1 or scFv2) comprises, from the N-terminus to the C-terminus: VL-optional linker-VH.

[0249] connector

[0250] In some embodiments, the isolated anti-human FD antibody construct includes one or more linkers between the anti-human FD antibody portion (e.g., anti-FDscFv) and the second antibody portion (e.g., scFv) that specifically recognizes complement proteins. The length, flexibility, and / or other properties of the linker include, but are not limited to, affinity, specificity, or affinity for one or more specific antigens or epitopes. For example, a longer linker may be chosen to ensure that two adjacent domains do not spatially interfere with each other. In some embodiments, the linker (e.g., a peptide linker) includes flexible residues (e.g., glycine (G) and serine (S)) such that the anti-human FD antibody portion (e.g., scFv) and the second antibody portion that specifically recognizes complement proteins are freely movable relative to each other. In some embodiments, the linker is a peptide linker. In some embodiments, the linker is a non-cleavable linker. In some embodiments, the linker is a cleavable linker.

[0251] Other linker considerations include the impact on the physical or pharmacokinetic properties of the resulting compound, such as solubility, lipophilicity, hydrophilicity, hydrophobicity, stability (various degrees of stability and planned degradation), rigidity, flexibility, immunogenicity, regulation of antibody binding, and ability to be incorporated into micelles or liposomes.

[0252] Peptide linkers can have naturally occurring or non-natural sequences. For example, sequences derived from the hinge region of heavy-chain-only antibodies can be used as linkers. See, for example, WO1996 / 034103.

[0253] The peptide linker can have any suitable length. In some embodiments, the length of the peptide linker is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 75, 100, 200 amino acids or more. In some embodiments, the length of the peptide linker is no more than about 200, 100, 75, 50, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5 amino acids or fewer. In some embodiments, the length of the peptide linker is any of the following: about 1 amino acid to about 10 amino acids, about 1 amino acid to about 20 amino acids, about 1 amino acid to about 30 amino acids, about 5 amino acids to about 15 amino acids, about 10 amino acids to about 25 amino acids, about 5 amino acids to about 30 amino acids, about 10 amino acids to about 30 amino acids, about 30 amino acids to about 50 amino acids, about 50 amino acids to about 100 amino acids, or about 1 amino acid to about 100 amino acids.

[0254] In some embodiments, the peptide linker does not contain any polymerization activity. Peptide linkers are known in the art to contain no promotion of secondary structure, and their characteristics are described, for example, by Dall'Acqua et al. (Biochem. (1998) 37, 9266-9273), Cheadle et al. (Mol Immunol (1992) 29, 21-30), and Raag and Whitlow (FASEB (1995) 9(1), 73-80). In some embodiments, the peptide linker does not promote the formation of any secondary structure. The bonding of the domains to each other can be provided, for example, by genetic engineering. Methods for preparing fused and operatively linked bispecific single-stranded constructs and expressing these constructs in mammalian cells or bacteria are well known in the art (e.g., WO1999 / 054440; Ausubel, Current Protocols in Molecular Biology, Green Publishing Associates and Wiley Interscience, NY 1989 and 1994; or Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2001).

[0255] Peptide linkers can be stable linkers that cannot be cleaved by proteases, especially matrix metalloproteinases (MMPs).

[0256] In some embodiments, the peptide linker is a flexible linker. Exemplary flexible linkers include glycine polymers (G)n, where n is at least one integer; and glycine-serine polymers (including, for example, (GS)n, where n is at least one integer); glycine-alanine polymers, alanine-serine polymers, and other flexible linkers known in the art. In some embodiments, the linker is a GS linker.

[0257] In some implementations, the isolated anti-human FD antibody construct (e.g., a multispecific construct) comprises two polypeptide chains, each containing a subunit of an Fc domain, and the two polypeptide chains are dimerized by the Fc domain.

[0258] In some embodiments, the isolated anti-human FD antibody construct containing the Fc domain (e.g., a multispecific construct) has one or more of the following properties: i) an in vivo half-life extended by at least about 20% (e.g., an extension of at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 1-fold, 1.5-fold, 2-fold, 5-fold, 10-fold or higher); and ii) an activity at least about 20% stronger (e.g., at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 1-fold, 1.5-fold, 2-fold, 5-fold, 10-fold or higher) than the same isolated anti-human FD antibody construct (e.g., a multispecific construct) without the Fc domain (e.g., different antibody moieties linked by a G4S linker).

[0259] In some embodiments, the anti-human FD antibody moiety is fused to the second antibody moiety via an Fc domain, such as an Fc domain containing the amino acid sequence of SEQ ID NO: 230. Any Fc domain described in the “Fc Domain” section above may be used in the linkers herein. In some embodiments, the Fc domain is derived from human IgG4 Fc. In some embodiments, the Fc domain is side-joined with GGGGS (SEQ ID NO: 221) at both ends of its sequence. In some embodiments, the Fc domain is side-joined with a linker (e.g., SEQ ID NO: 227) at one or both ends of its sequence and then located between the anti-human FD antibody moiety (e.g., anti-FD scFv1) and the second antibody moiety (e.g., scFv2). In some embodiments, the Fc domain is side-joined with a linker (e.g., SEQ ID NO: 227) at both ends of its sequence (hereinafter also referred to as “side-joined Fc domain linker”) and then located between the anti-human FD antibody moiety (e.g., anti-FD scFv1) and the second antibody moiety (e.g., scFv2). In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 230. In some embodiments, the side-attached Fc domain linker comprises the amino acid sequence of SEQ ID NO: 233.

[0260] In some embodiments, the anti-human FD antibody portion (e.g., scFv) is fused directly to a fusion complex or a second antibody portion (e.g., scFv). In some embodiments, the anti-human FD antibody portion (e.g., scFv) is fused to a fusion complex or a second antibody portion (e.g., scFv) via a linker (e.g., a peptide linker). In some embodiments, the linkers within the scFv (e.g., anti-human FD scFv and / or scFv targeting complement proteins) and / or the linkers between two or more antibody portions in the anti-human FD antibody construct are independently selected from the group consisting of SEQ ID NO: 221-229, such as any one of SEQ ID NO: 221 and 226-229.

[0261] In some embodiments, the isolated anti-human FD antibody construct comprises, from the N-terminus to the C-terminus: an anti-human FD scFv (e.g., any anti-human FD scFv described herein), an optional linker, and an anti-C2 scFv or an anti-C5 scFv. In some embodiments, the isolated anti-human FD antibody construct comprises, from the N-terminus to the C-terminus: an anti-C2 scFv or an anti-C5 scFv, an optional linker, and an anti-human FD scFv (e.g., any anti-human FD scFv described herein). In some embodiments, the isolated anti-human FD antibody construct comprises, from the N-terminus to the C-terminus: an anti-human FD scFv (e.g., any anti-human FD scFv described herein), a first optional linker, an Fc domain, a second optional linker, and an anti-C2 scFv or an anti-C5 scFv. In some embodiments, the isolated anti-human FD antibody construct comprises, from the N-terminus to the C-terminus: an anti-C2 scFv or an anti-C5 scFv, a first optional linker, an Fc domain, a second optional linker, and an anti-human FD scFv (e.g., any anti-human FD scFv described herein). In some embodiments, where the isolated anti-human FD antibody construct includes an Fc domain, either as a whole or as part of a linker, the two polypeptide chains, each containing an Fc subunit, dimerize through the Fc domain.

[0262] Complement factor 2 (C2)

[0263] In some embodiments, the complement factor bound to the second antibody moiety is complement factor 2 (C2), therefore the anti-human FD antibody construct is an anti-human FD / anti-C2 multispecific construct. Any anti-C2 antibody or its antigen-binding fragment described in WO2023143583 (the contents of which are incorporated herein by reference in their entirety) may be used in the anti-C2 antibody moiety described herein. In some embodiments, the anti-C2 antibody moiety is an anti-C2 scFv.

[0264] Human C2 is an 83 kDa protein (100 kDa after glycosylation) produced as an inactive, highly glycosylated zymogen, consisting of five domains: three N-terminal complement control protein (CCP) domains, one von Willebrand factor A (VWA) domain, and one C-terminal trypsin-like serine protease (SP) domain. The last two domains, VWA and SP, form the C2a fragment (residues 224–732, 70 kDa), which is generated in the canonical and lectin-binding proteolytic activation cascade. The larger C2a fragment is 57.4 kDa (70 kDa after glycosylation) and provides the catalytic center for the invertase complex of the complement-activated canonical and lectin-binding pathways. C2, bound to C4b, is cleaved by classical proteases (C1s) or lectin proteases (MASP2) to produce C4bC2a. C4bC2a is a short-lived C3 convertase that further cleaves C3 into C3a and C3b, ultimately leading to the formation of the membrane attack complex (MAC) and the lysis of bacteria and damaged cells. C2 shares the same serine protease domain as C4bC2a, but exhibits an inactive, zymogeneous conformation, requiring a cofactor-induced conformational change to exert its activity.

[0265] The anti-C2 antibody portion can specifically bind to C2 from any source, such as any organism having a complement system, including but not limited to dogs, cats, pigs, cattle, sheep, goats, horses, rats, rabbits, hamsters, guinea pigs, monkeys, mice, and humans. In some embodiments, C2 is mouse C2. In some embodiments, C2 is human C2. In some embodiments, C2 is cynomolgus monkey C2.

[0266] In some embodiments, the anti-C2 antibody portion binds to C2. In some embodiments, the anti-C2 antibody portion binds to C2a. In some embodiments, the anti-C2 antibody portion exhibits cross-species reactivity with C2 other than human C2 and / or with C2a other than human C2a. Exemplary non-human C2s and C2a include, but are not limited to, mouse C2 and C2a, rat C2 and C2a, rabbit C2 and C2a, sheep C2 and C2a, and cynomolgus monkey C2 and C2a. In some embodiments, the anti-C2 antibody portion cross-reacts with cynomolgus monkey C2 and / or C2a.

[0267] The anti-C2 antibody moiety can be any suitable form known in the art. In some embodiments, the anti-C2 antibody moiety is selected from the group consisting of: full-length antibodies, Fab, Fab', F(ab')2, scFv, sdAb, and combinations thereof. In some embodiments, the anti-C2 antibody moiety comprises an sdAb that binds to C2. In some embodiments, the anti-C2 antibody moiety is a full-length antibody, such as a full-length antibody comprising an Fc fragment derived from IgG4 (e.g., an IgG4 Fc containing a PLA mutation, such as SEQ ID NO:230). In some embodiments, the anti-C2 antibody moiety comprises anti-C2 scFv (or consists substantially of anti-C2 scFv, or consists of anti-C2 scFv). In some embodiments, the anti-C2 antibody moiety is a mouse antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the anti-C2 antibody moiety is monospecific. In some embodiments, the anti-C2 antibody moiety is multispecific. In some embodiments, the anti-C2 antibody moiety is monovalent. In some embodiments, the anti-C2 antibody moiety is multivalent.

[0268] In some embodiments, the binding of the anti-C2 antibody moiety to C2 (e.g., human C2) is pH-dependent, and the binding of the anti-C2 antibody moiety to C2 at neutral pH (e.g., about pH 7.4; e.g., the pH found in blood) is stronger than its binding at acidic pH (e.g., about pH 5.8; e.g., the pH found in endocrine systems). In some embodiments, the binding affinity of the pH-dependent anti-C2 antibody moiety to C2 (e.g., human C2) at about pH 7.4 is at least about 3 times (e.g., at least about 4, 5, 6, 7, 8, 9, 10 times or higher) than the binding affinity of the pH-dependent anti-C2 antibody moiety to C2 (e.g., human C2) at about pH 5.8.

[0269] In some embodiments, the anti-C2 antibody portion comprises: (i) VH, said VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 259, H-CDR2 containing the amino acid sequence of SEQ ID NO: 260, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 261; and (ii) VL, said VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 262, L-CDR2 containing the amino acid sequence of SEQ ID NO: 263, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 264. In some embodiments, the anti-C2 antibody portion comprises: a VH containing the amino acid sequence of SEQ ID NO: 265 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 97%, 99% or higher) amino acid homology with SEQ ID NO: 265; and a VL containing the amino acid sequence of SEQ ID NO: 266 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 97%, 99% or higher) amino acid homology with SEQ ID NO: 266. In some embodiments, the anti-C2 antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 265 and a VL containing the amino acid sequence of SEQ ID NO: 266. In some embodiments, the anti-C2 antibody portion is an scFv, such as an anti-C2 scFv containing the amino acid sequence of SEQ ID NO: 267 or 268.

[0270] In some embodiments, the anti-C2 antibody portion comprises: (i) VH, said VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 269, H-CDR2 containing the amino acid sequence of SEQ ID NO: 270, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 271; and (ii) VL, said VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 272, L-CDR2 containing the amino acid sequence of SEQ ID NO: 273, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 274. In some embodiments, the anti-C2 antibody portion comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 275 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 97%, 99% or higher) amino acid homology with SEQ ID NO: 275; and a VL domain comprising the amino acid sequence of SEQ ID NO: 276 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 97%, 99% or higher) amino acid homology with SEQ ID NO: 276. In some embodiments, the anti-C2 antibody portion comprises a VH containing the amino acid sequence of SEQ ID NO: 275 and a VL containing the amino acid sequence of SEQ ID NO: 276. In some embodiments, the anti-C2 antibody portion is an scFv, such as an anti-C2 scFv comprising the amino acid sequence of SEQ ID NO: 277 or 278.

[0271] In some implementations, the anti-C2 antibody partially and specifically binds to C2 (e.g., human C2), where K D It is about 10 -7 M to approximately 10 -12 M, for example, any of the following: approximately 10 -7 M to approximately 10 -10 M, approximately 10 -8 M to approximately 10 -11 M, approximately 10 -9 M to approximately 10 -11 M, approximately 10 -8 M to approximately 10 -10 M, approximately 10 -8 M to approximately 10 -9 M, approximately 10 -9 M to approximately 10 -10 M or approximately 10 -10 M to approximately 10 -11 M. In some embodiments, the anti-C2 antibody partially and specifically binds to C2 (e.g., human C2), where KD It is approximately 1×10 -10 M to approximately 5 × 10 -10 M.

[0272] In some embodiments, the anti-C2 antibody described herein partially inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or higher) CP and / or LP. In some embodiments, the anti-C2 antibody described herein partially inhibits no more than about 70% (e.g., no more than about 60%, 50%, 40%, 30%, 20%, 10% or lower) CP and / or LP. In some embodiments, the anti-C2 antibody partially does not inhibit (or inhibits at most about 10%, 5%, 2%, 1% or lower) AP activation. In some embodiments, the anti-C2 antibody partially inhibits about 50% to about 80% of CP and / or LP. In some embodiments, the anti-C2 antibody partially inhibits the IC50 of AP (e.g., inhibits rabbit RBC lysis in 20% human serum). 50 The value could not be detected. In some embodiments, the anti-C2 antibody partially inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) IgM-induced and / or mannan-induced C3b and / or C5b-9 deposition. In some embodiments, the anti-C2 antibody partially does not inhibit (or inhibits at most about 10%, 5%, 2%, 1%, or less) LPS-induced C3b and / or C5b-9 deposition.

[0273] In some embodiments, the anti-human FD / anti-C2 multispecific construct: i) inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) CP and LP, e.g., inhibits about 50% to about 80% of CP and / or LP, and ii) inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) AP. In some embodiments, the anti-human FD / anti-C2 multispecific construct inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) all of CP, LP, AP, and TP. In some embodiments, the anti-human FD / anti-C2 multispecific construct inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) IgM-induced, mannan-induced, and LPS-induced C3b and C5b-9 deposition. In some embodiments, the anti-human FD / anti-C2 multispecific construct inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) sheep RBC lysis and rabbit RBC lysis (e.g., induced by human serum).

[0274] Complement factor 5 (C5)

[0275] In some embodiments, the complement factor bound to the second antibody portion is complement factor 5 (C5), thus the anti-human FD antibody construct is an anti-human FD / anti-C5 multispecific construct. Any anti-C5 antibody or its antigen-binding fragment described in US20220204602, US20220177556, US11578137, WO2022134047, and PCT / US2023 / 063305 may be used in the anti-C5 antibody portion described herein, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the anti-C5 antibody portion is an anti-C5 scFv.

[0276] The C5 gene encodes a protoprotein that is proteased to produce various protein products, including the C5 α chain, C5 β chain, C5a anaphylatoxin, and C5b. Human C5 is a 188 kDa protein containing a C5 α chain and a β chain linked by disulfide bonds. Invertase cleaves the α chain, forming the potent anaphylatoxin C5a and the large C5b cleavage product, which possess strong convulsant and chemotactic activities. The C5b large cleavage product can form a complex with complement component C6, which then becomes the membrane attack complex (MAC). Mutations in this gene cause complement component 5 deficiency, a disease characterized by recurrent bacterial infections.

[0277] The anti-C5 antibody moiety can specifically bind to C5 from any source, such as any organism having a complement system, including but not limited to dogs, cats, pigs, cattle, sheep, goats, horses, rats, rabbits, hamsters, guinea pigs, monkeys, mice, and humans. In some embodiments, C5 is mouse C5. In some embodiments, C5 is human C5.

[0278] In some embodiments, the anti-C5 antibody portion binds to C5. In some embodiments, the anti-C5 antibody portion binds to C5b. In some embodiments, the anti-C5 antibody portion exhibits cross-species reactivity with C5s other than human C5 and / or with C5b other than human C5b. Exemplary non-human C5s and C5b include, but are not limited to, mouse C5 and C5b, rat C5 and C5b, rabbit C5 and C5b, sheep C5 and C5b, and cynomolgus monkey C5 and C5b. In some embodiments, the anti-C5 antibody portion cross-reacts with cynomolgus monkey C5 and / or C5b. In some embodiments, the anti-C5 antibody portion does not bind to C5a.

[0279] The anti-C5 antibody moiety can be any suitable form known in the art. In some embodiments, the anti-C5 antibody moiety is selected from the group consisting of: full-length antibodies, Fab, Fab', F(ab')2, scFv, sdAb, and combinations thereof. In some embodiments, the anti-C5 antibody moiety comprises an sdAb that binds to C5. In some embodiments, the anti-C5 antibody moiety is a full-length antibody, such as a full-length antibody comprising an Fc fragment derived from IgG4 (e.g., an IgG4 Fc containing a PLA mutation, such as SEQ ID NO:230). In some embodiments, the anti-C5 antibody moiety comprises anti-C5 scFv (or consists essentially of anti-C5 scFv, or consists of anti-C5 scFv). In some embodiments, the anti-C5 antibody moiety is a mouse antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the anti-C5 antibody moiety is monospecific. In some embodiments, the anti-C5 antibody moiety is multispecific. In some embodiments, the anti-C5 antibody moiety is monovalent. In some implementations, the anti-C5 antibody portion is multivalent.

[0280] In some embodiments, the binding of the anti-C5 antibody moiety to C5 (e.g., human C5) is pH-dependent, and the binding of the anti-C5 antibody moiety to C5 at neutral pH (e.g., about pH 7.4; e.g., the pH found in blood) is stronger than its binding at acidic pH (e.g., about pH 5.8; e.g., the pH found in endocrine systems). In some embodiments, the binding affinity of the pH-dependent anti-C5 antibody moiety to C5 (e.g., human C5) at about pH 7.4 is at least about 3 times (e.g., at least about 4, 5, 6, 7, 8, 9, 10 times or more) than the binding affinity of the pH-dependent anti-C5 antibody moiety to C5 (e.g., human C5) at about pH 5.8.

[0281] In some implementations, the anti-C5 antibody portion binds to an epitope in the α chain of C5 and / or an epitope in the β chain of C5.

[0282] In some embodiments, the anti-C5 antibody portion comprises: (i) VH, said VH comprising H-CDR1 containing the amino acid sequence of SEQ ID NO: 279, H-CDR2 containing the amino acid sequence of SEQ ID NO: 280, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 281; and (ii) VL, said VL comprising L-CDR1 containing the amino acid sequence of SEQ ID NO: 282, L-CDR2 containing the amino acid sequence of SEQ ID NO: 283, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 284. In some embodiments, the anti-C5 antibody moiety comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 285 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 97%, 99% or higher) amino acid homology with SEQ ID NO: 285; and a VL domain comprising the amino acid sequence of SEQ ID NO: 286 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 97%, 99% or higher) amino acid homology with SEQ ID NO: 286. In some embodiments, the anti-C5 antibody moiety comprises a VH containing the amino acid sequence of SEQ ID NO: 285 and a VL containing the amino acid sequence of SEQ ID NO: 286. In some embodiments, the anti-C5 antibody moiety is an scFv containing the amino acid sequence of SEQ ID NO: 287 or 288.

[0283] In some implementations, the anti-C5 antibody partially and specifically binds to C5 (e.g., human C5), where K D It is about 10 -7 M to approximately 10 -12 M, for example, any of the following: approximately 10 -7 M to approximately 10 -10 M, approximately 10 -8 M to approximately 10 -11 M, approximately 10 -9 M to approximately 10 -11 M, approximately 10 -8 M to approximately 10 -10 M, approximately 10 -8 M to approximately 10 -9 M, approximately 10 -9 M to approximately 10 -10 M or approximately 10 -10 M to approximately 10 -11 M. In some implementations, the anti-C5 antibody partially and specifically binds to C5 (e.g., human C5), where K DIt is approximately 1×10 -9 M to approximately 5 × 10 -9 M.

[0284] In some embodiments, the anti-human FD / anti-C5 multispecific construct: i) inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) TP, and ii) inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) AP. In some embodiments, the anti-human FD / anti-C5 multispecific construct inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) IgM-induced, mannan-induced, and LPS-induced C5b-9 deposition. In some embodiments, the anti-human FD / anti-C5 multispecific construct inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) LPS-induced C3b deposition. In some embodiments, the anti-human FD / anti-C5 multispecific construct does not inhibit (or inhibits at most about 10%, 5%, 2%, 1%, or less) IgM-induced or mannan-induced C3b deposition. In some embodiments, the anti-FD / anti-C5 multispecific construct inhibits (e.g., inhibits at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) sheep RBC lysis and rabbit RBC lysis (e.g., induced by human serum).

[0285] In some embodiments, the complement pathway activity inhibited by the anti-human FD / anti-C5 multispecific construct is complement pathway activation induced by one or more of LPS, LOS, PAMP, and DAMP. In some embodiments, the complement signaling activity inhibited by the anti-human FD / anti-C5 multispecific construct is the production of C5a protein, the production of C5b protein, and / or the formation of MAC. In some embodiments, the complement pathway activity inhibited by the anti-human FD / anti-C5 multispecific construct is C5-dependent.

[0286] The activity assays for the multispecific constructs described herein are well known in the art, including, for example, inhibition of one or more of AP, CP, LP, and TP, including but not limited to, rabbit RBC lysis assays (e.g., induced by human serum), sheep RBC lysis assays (e.g., induced by human serum), IgM-induced C3b and / or C5b-9 deposition assays, mannan-induced C3b and / or C5b-9 deposition assays, or LPS-induced C3b and / or C5b-9 deposition assays. See, for example, the assays in US11434279, WO2023143583, US20220204602, US20220177556, US11578137, WO2022134047, and PCT / US2023 / 063305, the contents of which are incorporated herein by reference in their entirety. See also Example 4 herein.

[0287] Chimeric antibodies or humanized antibodies

[0288] In some embodiments, the isolated anti-human FD antibody construct (e.g., a multispecific construct) or one or more antibody portions of the anti-human FD antibody moiety of this application are chimeric antibodies. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and in Morrison et al. Proc. Natl. Acad. Sci. USA In 81:6851-6855 (1984), chimeric antibodies comprise a non-human variable region (e.g., a variable region derived from a mouse) and a human constant region. In some embodiments, chimeric antibodies are "class-switched" antibodies, where the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include their antigen-binding fragments.

[0289] In some embodiments, the chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce its immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more variable domains, wherein the HVR, such as the CDR (or a portion thereof), is derived from the non-human antibody, and the FR (or a portion thereof) is derived from the human antibody sequence. Optionally, the humanized antibody will also contain at least a portion of the human constant region. In some embodiments, some FR residues in the humanized antibody are substituted with corresponding residues from the non-human antibody (e.g., the HVR residues are derived from the antibody therein) to, for example, restore or enhance antibody specificity or affinity.

[0290] Humanized antibodies and their preparation methods have been reviewed, for example, by Almagro and Fransson. Front. Biosci.13:1619-1633 (2008), and further described in, for example, Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (Description of SDR (a-CDR) porting); Padlan, Mol. Immunol. 28:489-498 (1991) (Description of “Surface Remodeling”); Dall'Acqua et al., Methods 36:43-60 (2005) (describing “FR reorganization”); and Osbourn et al., Methods 36:61-68 (2005) and Klimka, Br. J. Cancer , 83:252-260 (2000) (Describes the “guided selection” approach to FR reorganization).

[0291] Human frame regions that can be used for humanization include, but are not limited to: frame regions selected using the "best-fit" method (see, for example, Sims et al.). J. Immunol 15 1 :2296 (1993)); Frame regions derived from common sequences of specific subgroups of the variable regions of human antibody light or heavy chains (see, for example, Carter et al., Proc. Natl. Acad. Sci. USA , 8 9 :4285 (1992); and Presta et al., J. Immunol. , 151:2623 (1993)); human maturation (somatic mutation) framework region or human germline framework region (see, for example, Almagro and Fransson, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and the frame regions obtained by screening FR libraries (see, for example...). Baca people, J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0292] Human antibodies

[0293] In some embodiments, one or more antibody portions of the isolated anti-human FD antibody construct (e.g., a multispecific construct) of this application are human antibodies (referred to as human domain antibodies, or human dAbs). Human antibodies can be generated using various techniques known in the art. Human antibodies are broadly described in van Dijk and van de Winkel. Curr. Opin. Pharmacol. 5: 368-74 (2001), Lonberg, Curr. Opin. Immunol. 20:450-459 (2008) and Chen, Mol. Immunol. In 47(4):912-21 (2010). Transgenic mice or rats capable of producing fully human single-domain antibodies (or dAbs) are known in the art. See, for example, US20090307787A1, US Patent No. 8,754,287, US20150289489A1, US20100122358A1 and WO2004049794.

[0294] Human antibodies (e.g., human dAbs) can be prepared by administering an immunogen to transgenic animals modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigen stimulation. Such animals typically contain all or part of the human immunoglobulin loci, which replace endogenous immunoglobulin loci or are located extrachromosomally or randomly integrated into the animal's chromosome. In such transgenic mice, endogenous immunoglobulin loci are usually inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg. Nat. Biotech. 23:1117-1125 (2005). See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE™ technology; and U.S. Patent No. 5,770,429, which describes HuMab. ® The technology is described in U.S. Patent No. 7,041,870 and in U.S. Patent Application Publication No. US 2007 / 0061900. The human variable region derived from intact antibodies produced by such animals can be further modified, for example, by combining it with different human constant regions.

[0295] Human antibodies (e.g., human dAbs) can also be prepared using hybridoma-based methods. Human myeloma and mouse-human xenograft cell lines for generating human monoclonal antibodies have been described (see, for example, Kozbor). J. Immunol. ,133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and ApplicationsPages 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al. J. Immunol ., 147: 86 (1991)). Human antibodies produced via human B-cell hybridoma technology were also described by Li et al., Proc. Natl. Acad. Sci. USA , 103:3557-3562 (2006). Additional methods include those described, for example, in U.S. Patent No. 7,189,826 (description of generating monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue , 26(4):265-268 (2006) (Description of those methods for human-human hybridomas). Human hybridoma technology (tri-source hybridoma technology) is also described by Vollmers and Brandlein, Histology and Histopathology , 20(3):927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology , 27(3):185-91 (2005)

[0296] Human antibodies (e.g., human dAbs) can also be generated by isolating variable domain sequences from Fv clones selected from a human phage display library. These variable domain sequences can then be combined with desired human constant domains.

[0297] Replacement, insertion, and missing variants

[0298] In some embodiments, the isolated anti-human FD antibody construct (e.g., a multispecific construct) or anti-human FD antibody moiety described herein includes an antibody variant containing one or more amino acid substitutions. Sites of interest for substitution mutagenesis include HVR (or CDR) and FR. Conserved substitutions are shown in... Table B Under the heading "Preferred Substitutions," the most significant changes are provided. Table B Under the heading “Exemplary Substitutions”, and further described below with reference to the amino acid side chain category, amino acid substitutions can be introduced into antibodies of interest, and products can be screened for desired activities, such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[0299] Table B. Amino Acid Substitution

[0300] Amino acids can be grouped according to the properties of their common side chains: (1) hydrophobic: leucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that affect chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[0301] Non-conservative replacement would require replacing members of one of these categories with members of another category.

[0302] One type of substitution variant involves replacing one or more hypervariable residues of a parent antibody (e.g., a humanized or human antibody). Generally, the resulting variants selected for further research will have modifications (e.g., improvements) to certain biological properties relative to the parent antibody (e.g., increased affinity, decreased immunogenicity) and / or will retain some biological properties of the parent antibody substantially. An exemplary substitution variant is an affinity-matured antibody, which can be conveniently generated, for example, using phage display-based affinity maturation techniques, such as those described herein. In short, one or more HVR residues are mutated, causing the variant antibody to be displayed on a phage and screened for specific biological activities (e.g., binding affinity).

[0303] HVR can be modified (e.g., substituted), for example, to improve antibody affinity. HVR "hotspots," which are residues encoded by codons that undergo mutations at a high frequency during somatic cell maturation (see, for example, Chowdhury), can also be modified. Methods Mol. Biol . 207:179-196 (2008)), and / or SDR (a-CDR) make such changes, wherein the resulting variant VH or VL is tested for binding affinity. Affinity maturation via the construction and reselection of secondary libraries has been described, for example, by Hoogenboom et al. Methods in Molecular Biology 178:1-37 (O'Brien et al., eds., Human Press, Totowa, NJ, (2001)). In some embodiments of affinity maturation, diversity is introduced into the variable gene for maturation selection by any of a variety of methods (e.g., error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity. Another approach to introducing diversity involves an HVR-directed pathway, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding can be specifically identified, for example, using alanine scan mutagenesis or modeling. Specifically, CDR-H3 and CDR-L3 are often targeted.

[0304] In some implementations, substitutions, insertions, or deletions can occur within one or more HVRs, as long as such changes do not materially reduce the antibody's ability to bind to the antigen. For example, conserved changes to the HVR that do not materially reduce binding affinity can be made (e.g., conserved substitutions as described herein). Such changes can be made outside the HVR "hotspot" or CDR. In some implementations of the variant VHH sequences provided above, each HVR is either unchanged or contains no more than one, two, or three amino acid substitutions.

[0305] One method that can be used to identify residues or regions in antibodies that can serve as mutagenic targets is called "alanine scanning mutagenesis," as described by Cunningham and Wells (1989). Science As described in 244:1081-1085. In this method, groups of residues or target residues (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) are identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the antibody-antigen interaction is affected. Other substitutions may be introduced at amino acid positions that show functional sensitivity to the initial substitution. Alternatively or additionally, the crystal structure of the antigen-antibody complex is used to identify the contact points between the antibody and antigen. Such contact residues and adjacent residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they contain the desired properties.

[0306] Amino acid sequence insertions include N-terminal and / or C-terminal fusions of peptides ranging in length from one residue to one hundred or more residues, as well as intra-sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies having an N-terminal methionyl residue. Other insertion variants of antibody molecules include fusions of the N-terminus or C-terminus of the antibody with an enzyme (e.g., for ADEPT) or a peptide that extends the serum half-life of the antibody.

[0307] Glycosylation variants

[0308] In some embodiments, one or more antibody portions of the isolated anti-human FD antibody construct (e.g., a multispecific construct) or the anti-human FD antibody moiety of this application are modified to increase or decrease the degree of glycosylation of said construct. The addition or deletion of glycosylation sites in the antibody can be conveniently achieved by altering the amino acid sequence to generate or remove one or more glycosylation sites.

[0309] When an antibody moiety (or isolated anti-human FD antibody construct) contains an Fc domain, the carbohydrate linked to it can be altered. Naturally occurring antibodies produced by mammalian cells typically contain branched, biantennary oligosaccharides, generally linked to the C-terminal of the Fc domain via N-bonds.H 2. Asn297 of the structural domain. See, for example, Wright et al. TIBTECH 15:26-32 (1997). Oligosaccharides can include various carbohydrates such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to the GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some embodiments, the oligosaccharides in the antibody moiety can be modified to produce antibody variants with certain improved properties.

[0310] In some embodiments, the antibody moiety (or isolated anti-human FD antibody construct) has a carbohydrate structure lacking fucose linked (directly or indirectly) to the Fc domain. For example, the amount of fucose in such antibodies can be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose at Asn297 within the sugar chain relative to the sum of all sugar structures linked to Asn297 (e.g., complex, heterogeneous, and high-mannose structures), measured by MALDI-TOF mass spectrometry, as described, for example, in WO 2008 / 077546. Asn297 refers to the asparagine residue located at approximately position 297 (EU number of the Fc domain residue) in the Fc domain; however, Asn297 may also be located approximately ±3 amino acids upstream or downstream of position 297 due to minor sequence variations in the antibody, i.e., between position 294 and position 300. Such fucosylated variants can have improved ADCC function. See, for example, U.S. Patent Publication Nos. US2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd.). Publicly disclosed examples involving "defucosylated" or "fucosylated" antibody variants include: US 2003 / 0157108; WO2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng.87: 614 (2004). Examples of cell lines capable of producing defucosylation antibodies include Lec13 CHO cells with protein fucosylation defects (Ripka et al., Arch. Biochem. Biophys 249:533-545 (1986); US Patent Application No. US 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., especially Example 11); and gene knockout cell lines, such as α-1,6-fucosyltransferase gene. FUT8 Gene knockout CHO cells (see, for example, Yamane-Ohnuki et al., Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng ., 94(4):680-688 (2006); and WO2003 / 085107).

[0311] In some embodiments, the antibody moiety (or isolated anti-human FD antibody construct) has a bisected oligosaccharide, for example, wherein the biantennary oligosaccharide linked to the antibody Fc domain is bisected via GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and US 2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide linked to the Fc domain are also provided. Such antibody variants may have improved CDC function. This antibody variant is described, for example, in WO1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0312] Cysteine-engineered antibody variants

[0313] In some embodiments, it may be desirable to produce cysteine-engineered antibody moieties, such as "thioMAb," wherein one or more residues of one or more antibody moieties in the isolated anti-FD antibody construct described herein are replaced by cysteine ​​residues. In some embodiments, the substituted residues are located at accessible sites on the antibody. By replacing those residues with cysteine, a reactive thiol group is thereby positioned at an accessible site on the antibody and can be used to conjugate the antibody to other moieties (e.g., pharmaceutical moieties or linker-pharmaceutical moieties) to produce immunoconjugates, as further described herein. In some embodiments, cysteine ​​may be used to replace any or more of the following residues: A118 (EU number) of the heavy chain; and S400 (EU number) of the Fc domain of the heavy chain. Cysteine-engineered antibody moieties can be produced, for example, as described in U.S. Patent No. 7,521,541.

[0314] III. Nucleic Acids, Vectors, and Host Cells

[0315] Nucleic acid molecules encoding any isolated anti-human FD antibody construct (e.g., an anti-human FD antibody moiety, an anti-human FD / anti-C2 multispecific construct, or an anti-human FD / anti-C5 multispecific construct) or any of the various antibody moieties described herein are also covered herein. In some embodiments, a nucleic acid (e.g., an isolated nucleic acid) is provided that encodes an anti-human FD antibody construct or various antibody moieties (e.g., one or more polypeptide components thereof). mRNA comprising such nucleic acids is also provided. In some embodiments, the mRNA is formulated in liposomes or lipid nanoparticles. Vectors comprising any of the nucleic acids described herein (e.g., isolated nucleic acids), such as viral vectors (e.g., AAV vectors or lentiviral vectors), are also provided.

[0316] This also includes isolated host cells containing or expressing any of the isolated anti-human FD antibody constructs described herein (e.g., anti-human FD antibody moiety, anti-human FD / anti-C2 multispecific construct, or anti-human FD / anti-C5 multispecific construct) (e.g., one or more polypeptide components thereof). Isolated host cells containing nucleic acids encoding an isolated anti-human FD antibody construct described herein (e.g., one or more polypeptide components thereof) are also provided. Isolated host cells comprising a vector containing nucleic acids encoding an isolated anti-human FD antibody construct described herein (e.g., one or more polypeptide components thereof) are also provided.

[0317] This application also includes variants of these nucleic acid sequences. For example, the variants include nucleotide sequences that hybridize under at least moderately stringent hybridization conditions to a nucleic acid sequence encoding an anti-human FD antibody construct (e.g., an anti-human FD antibody moiety, an anti-human FD / anti-C2 multispecific construct, or an anti-human FD / anti-C5 multispecific construct) or various antibody moieties (e.g., one or more polypeptide components thereof) isolated herein.

[0318] This application also provides a vector into which the nucleic acid of this application is inserted.

[0319] The nucleic acid can be cloned into various types of vectors. For example, the nucleic acid can be cloned into vectors, including but not limited to plasmids, phage particles, phage derivatives, animal viruses, and entrapments. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

[0320] Alternatively, expression vectors can be provided to cells in the form of viral vectors. Viral vector technology is well known in the art and described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and other virology and molecular biology manuals. Viruses that can be used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses. Generally, suitable vectors contain an origin of replication that functions in at least one organism, a promoter sequence, a convenient restriction endonuclease site, and one or more selectivity markers (see, for example, WO 01 / 96584; WO 01 / 29058; and U.S. Patent No. 6,326,193).

[0321] In some embodiments, the nucleic acid encoding the isolated anti-human FD antibody construct (e.g., an anti-human FD antibody moiety, an anti-human FD / anti-C2 multispecific construct, or an anti-human FD / anti-C5 multispecific construct) contains, in addition to the coding sequence encoding the isolated anti-human FD antibody construct, other sequences or features. For example, the nucleic acid may be an mRNA (or a DNA sequence encoding the mRNA) containing optional features that improve the targeting of the isolated anti-human FD antibody construct encoded by the mRNA and enhance the stability of the mRNA.

[0322] In some embodiments, the mRNA encoding a portion or isolated anti-human FD antibody construct contains a nucleic acid sequence encoding a signal peptide. The signal peptide is a short peptide that may be present at the N-terminus or C-terminus of a newly synthesized protein and can be used for the correct translocation of the protein. In some embodiments, the signal peptide facilitates the translocation of the portion or isolated anti-human FD antibody construct encoded by the coding sequence of the mRNA. In some embodiments, the signal peptide translocates the portion or isolated anti-human FD antibody construct to the cell membrane.

[0323] In some embodiments, the mRNA contains one or more untranslated regions (UTRs). The UTRs of mRNA may be involved in various regulatory aspects of gene expression. It should be understood that the UTRs provided herein (e.g., 5' UTRs and / or 3' UTRs) are merely examples, and the mRNA can contain any UTR from any gene. Furthermore, multiple wild-type UTRs of any known gene can be utilized. The invention also aims to provide synthetic (e.g., artificial UTRs) that are not variants of wild-type genes and are also within the scope of the invention. These UTRs, or portions thereof, may be placed in the same orientation as in the transcripts in which they were selected, or their orientation or position may be altered. Thus, 5' or 3' UTRs may be inverted, shortened, lengthened, or form chimeras with one or more other 5' UTRs or 3' UTRs. As used herein, the term "alteration" in relation to a UTR sequence means that the UTR has undergone some change relative to a reference sequence. For example, a 3' or 5' UTR may be altered relative to a wild-type or native UTR as taught above by changing its orientation or position, or by including additional nucleotides, deleted nucleotides, exchanged nucleotides, or transposable nucleotides. Any of these variations that produce a “change” in the UTR (whether 3' or 5') constitutes a variant UTR.

[0324] In some implementations, dual, triple, or quadruple UTRs, such as 5' or 3' UTRs, may be used. As used herein, a "dual" UTR refers to a UTR in which two copies of the same UTR are tandemly or substantially tandemly encoded. The presence of patterned UTRs is also within the scope of this invention. As used herein, a "patterned UTR" is a UTR that reflects a repeating or alternating pattern, such as ABABAB or AABBBAABBAABB or ABCABCABC or variations thereof repeated once, twice, or more than three times. In these patterns, each letter A, B, or C represents a different UTR at the nucleotide level. UTRs provided herein may also include translation enhancer elements (TEEs).

[0325] In some embodiments, the mRNA contains a 5' UTR. The 5' UTR provided herein is recognized by ribosomes, enabling them to bind and initiate the translation of the mRNA (e.g., translation of the nucleic acid encoding the coding sequence and / or the signal peptide encoding the mRNA). In some embodiments, the 5' UTR is located upstream of the mRNA coding sequence.

[0326] In some embodiments, the mRNA contains a 3' UTR. The 3' UTR described herein can participate in translation termination (e.g., translation of the coding sequence of the mRNA and / or the nucleic acid encoding a signal peptide) and is also important for post-transcriptional modifications. In some embodiments, the 3' UTR is located upstream of the mRNA coding sequence. In some embodiments, the 3' UTR immediately follows a translation stop codon in the mRNA coding sequence. In some embodiments, the mRNA contains one or more stop codons preceding the 3' UTR.

[0327] In some embodiments, the mRNA comprises a 5' UTR and a 3' UTR, such as either the 5' UTR or 3' UTR provided herein. In some embodiments, the 5' UTR and 3' UTR are derived from the same species. In some embodiments, the 5' UTR and 3' UTR are not derived from the same species. In some embodiments, the 5' UTR is synthetic, while the 3' UTR is not synthetic. In some embodiments, the 5' UTR is not synthetic, while the 3' UTR is synthetic.

[0328] In some implementations, the mRNA contains one or more additional features, such as, but not limited to, a poly(A) sequence, one or more chemical modifications, a 5' cap, or a combination thereof.

[0329] In some embodiments, the mRNA includes a poly(A) sequence (e.g., a polyadenylated sequence). The poly(A) sequence consists of multiple consecutive adenosine monophosphates. In some embodiments, the poly(A) sequence is essential for the translation of the mRNA. In some embodiments, the poly(A) sequence is downstream of the mRNA coding sequence. In some embodiments, the poly(A) sequence is downstream of the 3' UTR of the mRNA. In some embodiments, the poly(A) sequence is about 50 nucleotides or longer, such as about 60, 70, 80, 90, 100, 150 nucleotides or longer. In some embodiments, the poly(A) sequence is about 150 nucleotides or shorter, such as about 100, 90, 80, 70, 50 or shorter. In some embodiments, the poly(A) sequence is about 105 nucleotides long.

[0330] Carrier and carrier delivery

[0331] This application also covers vectors comprising nucleic acids encoding any isolated anti-human FD antibody construct described herein (e.g., an anti-human FD antibody moiety, an anti-human FD / anti-C2 multispecific construct, or an anti-human FD / anti-C5 multispecific construct). As used herein, the term "vector" refers to a DNA or RNA molecule containing a nucleotide sequence that can be transcribed into RNA or expressed into a protein. In some embodiments, the vector contains one or more regulatory elements operatively linked to a nucleotide sequence encoding an isolated anti-human FD antibody construct. When the vector is introduced into a host cell, the nucleotide sequence encoding the construct can be transcribed or expressed under suitable conditions.

[0332] In some embodiments, the vector contains a promoter operatively linked to a coding nucleotide sequence such that the promoter controls the transcription or expression of the coding nucleotide sequence. The promoter may be located at the 5' (upstream) of the coding nucleotide sequence it controls. The distance between the promoter and the coding sequence may be approximately equal to the distance between the promoter in the gene from which the promoter originates and the gene it controls. As is known in the art, variations in this distance may be permissible without loss of promoter function. In some embodiments, the vector contains a 5' UTR and / or a 3' UTR for regulating the transcription or expression of the coding nucleotide sequence. In some embodiments, the promoter is a U6 promoter. In some embodiments, the promoter is the Poly II promoter discussed in the preceding sections.

[0333] Suitable vectors include, but are not limited to: single-stranded, double-stranded, or partially double-stranded nucleic acid molecules; nucleic acid molecules containing one or more free ends, or without free ends (e.g., circular); nucleic acid molecules containing DNA, RNA, or both; and other types of polynucleotides known in the art. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop in which additional DNA segments can be inserted, for example, by standard molecular cloning techniques. Some vectors are capable of autonomous replication in the host cell into which they are introduced (e.g., bacterial vectors with bacterial origins of replication and attachable mammalian vectors). Other vectors (e.g., non-attached mammalian vectors) integrate into the host cell's genome after introduction into the host cell and thereby replicate along with the host genome. Furthermore, some vectors are capable of guiding the transcription or expression of coding nucleotide sequences operatively linked to them. Such vectors are referred to herein as "expression vectors."

[0334] In some implementations, the expression vector is selected from the group consisting of ...

Claims

1. An isolated antibody construct (anti-human FD antibody construct) comprising an antibody moiety that specifically binds to human factor D (anti-human FD antibody moiety), wherein the anti-human FD antibody moiety comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), wherein: (i) The VH comprises: heavy chain CDR1 ("H-CDR1"), wherein the H-CDR1 comprises the amino acid sequence of SEQ ID NO: 1 or a variant thereof comprising up to 3 amino acid variations; heavy chain CDR2 ("H-CDR2"), wherein the H-CDR2 comprises the amino acid sequence of SEQ ID NO: 2 or a variant thereof comprising up to 3 amino acid variations; and heavy chain CDR3 ("H-CDR3"), wherein the H-CDR3 comprises the amino acid sequence of SEQ ID NO: 3 or a variant thereof comprising up to 3 amino acid variations; and (ii) The VL comprises: a light chain CDR1 ("L-CDR1"), the L-CDR1 comprising the amino acid sequence of SEQ ID NO: 4 or a variant thereof comprising up to 3 amino acid variations; a light chain CDR2 ("L-CDR2"), the L-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 or a variant thereof comprising up to 3 amino acid variations; and a light chain CDR3 ("L-CDR3"), the L-CDR3 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to 3 amino acid variations.

2. The isolated anti-human FD antibody construct as described in claim 1, wherein: (i) The VH comprises: H-CDR1 containing the amino acid sequence DTYVH (SEQ ID NO: 1); H-CDR2 containing the amino acid sequence RIDP-X1-X2-G-X3-T-X4-F-X5-PRFQA (SEQ ID NO: 9), wherein X1 is A or H, X2 is N, S or Y, X3 is L or H, X4 is T or H, and X5 is D, V, L or H; and H-CDR3 containing the amino acid sequence AMEY (SEQ ID NO: 3); and (ii) The VL comprises: L-CDR1 containing the amino acid sequence SA-X6-SDVS-X7-MY (SEQ ID NO: 10), wherein X6 is R, N or S and X7 is Y, D or V; L-CDR2 containing the amino acid sequence X8-TSNLAS (SEQ ID NO: 252), wherein X8 is D or H; and L-CDR3 containing the amino acid sequence QQWSS-X9-PPW-X10 (SEQ ID NO: 11), wherein X9 is Y or H and X10 is T, R or H.

3. The isolated anti-human FD antibody construct as described in claim 1 or 2, wherein: (1) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 1, H-CDR2 containing the amino acid sequence of SEQ ID NO: 2, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 4, L-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 6; (2) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 20, H-CDR2 containing the amino acid sequence of SEQ ID NO: 21, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 22; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 23, L-CDR2 containing the amino acid sequence of SEQ ID NO: 24, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 25; (3) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 36, H-CDR2 containing the amino acid sequence of SEQ ID NO: 37, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 38; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 39, L-CDR2 containing the amino acid sequence of SEQ ID NO: 40, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 41; (4) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 44, H-CDR2 containing the amino acid sequence of SEQ ID NO: 45, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 46; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 47, L-CDR2 containing the amino acid sequence of SEQ ID NO: 48, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 49; (5) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 52, H-CDR2 containing the amino acid sequence of SEQ ID NO: 53, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 54; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57; (6) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 63, L-CDR2 containing the amino acid sequence of SEQ ID NO: 64, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 65; (7) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 71, L-CDR2 containing the amino acid sequence of SEQ ID NO: 72, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 73; (8) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 76, H-CDR2 containing the amino acid sequence of SEQ ID NO: 77, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 78; the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 79, L-CDR2 containing the amino acid sequence of SEQ ID NO: 80, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 81; (9) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 84, H-CDR2 containing the amino acid sequence of SEQ ID NO: 85, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 86; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89; (10) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 92, H-CDR2 containing the amino acid sequence of SEQ ID NO: 93, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 94; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97; (11) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 100, H-CDR2 containing the amino acid sequence of SEQ ID NO: 101, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 102; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (12) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 108, H-CDR2 containing the amino acid sequence of SEQ ID NO: 109, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 110; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113; (13) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 116, H-CDR2 containing the amino acid sequence of SEQ ID NO: 117, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 118; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (14) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126; (15) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126; (16) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 4, L-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 6; (17) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 1, H-CDR2 containing the amino acid sequence of SEQ ID NO: 2, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 3; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 124, L-CDR2 containing the amino acid sequence of SEQ ID NO: 125, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 126; (18) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89; (19) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (20) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97; (21) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113; (22) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 128, H-CDR2 containing the amino acid sequence of SEQ ID NO: 129, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 130; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (23) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57; (24) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89; (25) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (26) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97; (27) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113; (28) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 60, H-CDR2 containing the amino acid sequence of SEQ ID NO: 61, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 62; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (29) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 55, L-CDR2 containing the amino acid sequence of SEQ ID NO: 56, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 57; (30) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 87, L-CDR2 containing the amino acid sequence of SEQ ID NO: 88, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 89; (31) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 103, L-CDR2 containing the amino acid sequence of SEQ ID NO: 104, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 105; (32) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 95, L-CDR2 containing the amino acid sequence of SEQ ID NO: 96, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 97; (33) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 111, L-CDR2 containing the amino acid sequence of SEQ ID NO: 112, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 113; (34) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 68, H-CDR2 containing the amino acid sequence of SEQ ID NO: 69, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 70; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 119, L-CDR2 containing the amino acid sequence of SEQ ID NO: 120, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 121; (35) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 206, H-CDR2 containing the amino acid sequence of SEQ ID NO: 207, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 208; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 209, L-CDR2 containing the amino acid sequence of SEQ ID NO: 210, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 211; (36) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 234, H-CDR2 containing the amino acid sequence of SEQ ID NO: 235, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 236; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 237, L-CDR2 containing the amino acid sequence of SEQ ID NO: 238, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 239; (37) The VH comprises H-CDR1 containing the amino acid sequence of SEQ ID NO: 240, H-CDR2 containing the amino acid sequence of SEQ ID NO: 241, and H-CDR3 containing the amino acid sequence of SEQ ID NO: 242; and the VL comprises L-CDR1 containing the amino acid sequence of SEQ ID NO: 243, L-CDR2 containing the amino acid sequence of SEQ ID NO: 244, and L-CDR3 containing the amino acid sequence of SEQ ID NO: 245; or (38) The VH includes H-CDR1 containing the amino acid sequence of SEQ ID NO: 246, H-CDR2 containing the amino acid sequence of SEQ ID NO: 247 and H-CDR3 containing the amino acid sequence of SEQ ID NO: 248; and the VL includes L-CDR1 containing the amino acid sequence of SEQ ID NO: 249, L-CDR2 containing the amino acid sequence of SEQ ID NO: 250 and L-CDR3 containing the amino acid sequence of SEQ ID NO:

251.

4. The isolated anti-human FD antibody construct according to any one of claims 1 to 3, wherein the VH comprises the amino acid sequence of SEQ ID NO: 7, or a variant thereof having at least about 80% amino acid sequence homology with SEQ ID NO: 7; and the VL comprises the amino acid sequence of SEQ ID NO: 8, or a variant thereof having at least about 80% amino acid sequence homology with SEQ ID NO:

8.

5. The isolated anti-human FD antibody construct as described in claim 4, wherein: i) The amino acid residue at position 24 of VH is A or T; ii) The amino acid residue at position 74 of VH is K or T; iii) The amino acid residue at position 77 of VH is N or S; iv) The amino acid residue at position 97 of VH is A or T; v) The amino acid residue at position 98 of the VH is R or Y; and / or vi) The amino acid residue at position 70 of VL is F or Y. The numbering is based on the Kabat numbering system.

6. The isolated anti-human FD antibody construct according to any one of claims 1 to 5, wherein: (1) The VH contains the amino acid sequence of SEQ ID NO: 7, and the VL contains the amino acid sequence of SEQ ID NO: 8; (2) The VH contains the amino acid sequence of SEQ ID NO: 18, and the VL contains the amino acid sequence of SEQ ID NO: 19; (3) The VH contains the amino acid sequence of SEQ ID NO: 26, and the VL contains the amino acid sequence of SEQ ID NO: 27; (4) The VH contains the amino acid sequence of SEQ ID NO: 34, and the VL contains the amino acid sequence of SEQ ID NO: 35; (5) The VH contains the amino acid sequence of SEQ ID NO: 42, and the VL contains the amino acid sequence of SEQ ID NO: 43; (6) The VH contains the amino acid sequence of SEQ ID NO: 50, and the VL contains the amino acid sequence of SEQ ID NO: 51; (7) The VH contains the amino acid sequence of SEQ ID NO: 58, and the VL contains the amino acid sequence of SEQ ID NO: 59; (8) The VH contains the amino acid sequence of SEQ ID NO: 66, and the VL contains the amino acid sequence of SEQ ID NO: 67; (9) The VH contains the amino acid sequence of SEQ ID NO: 74, and the VL contains the amino acid sequence of SEQ ID NO: 75; (10) The VH contains the amino acid sequence of SEQ ID NO: 82, and the VL contains the amino acid sequence of SEQ ID NO: 83; (11) The VH contains the amino acid sequence of SEQ ID NO: 90, and the VL contains the amino acid sequence of SEQ ID NO: 91; (12) The VH contains the amino acid sequence of SEQ ID NO: 98, and the VL contains the amino acid sequence of SEQ ID NO: 99; (13) The VH contains the amino acid sequence of SEQ ID NO: 106, and the VL contains the amino acid sequence of SEQ ID NO: 107; (14) The VH contains the amino acid sequence of SEQ ID NO: 114, and the VL contains the amino acid sequence of SEQ ID NO: 115; (15) The VH contains the amino acid sequence of SEQ ID NO: 122, and the VL contains the amino acid sequence of SEQ ID NO: 123; (16) The VH contains the amino acid sequence of SEQ ID NO: 156, and the VL contains the amino acid sequence of SEQ ID NO: 157; (17) The VH contains the amino acid sequence of SEQ ID NO: 158, and the VL contains the amino acid sequence of SEQ ID NO: 159; (18) The VH contains the amino acid sequence of SEQ ID NO: 160, and the VL contains the amino acid sequence of SEQ ID NO: 161; (19) The VH contains the amino acid sequence of SEQ ID NO: 162, and the VL contains the amino acid sequence of SEQ ID NO: 163; (20) The VH contains the amino acid sequence of SEQ ID NO: 164, and the VL contains the amino acid sequence of SEQ ID NO: 165; (21) The VH contains the amino acid sequence of SEQ ID NO: 166, and the VL contains the amino acid sequence of SEQ ID NO: 167; (22) The VH contains the amino acid sequence of SEQ ID NO: 168, and the VL contains the amino acid sequence of SEQ ID NO: 169; (23) The VH contains the amino acid sequence of SEQ ID NO: 172, and the VL contains the amino acid sequence of SEQ ID NO: 173; (24) The VH contains the amino acid sequence of SEQ ID NO: 174, and the VL contains the amino acid sequence of SEQ ID NO: 175; (25) The VH contains the amino acid sequence of SEQ ID NO: 176, and the VL contains the amino acid sequence of SEQ ID NO: 177; (26) The VH contains the amino acid sequence of SEQ ID NO: 178, and the VL contains the amino acid sequence of SEQ ID NO: 179; (27) The VH contains the amino acid sequence of SEQ ID NO: 180, and the VL contains the amino acid sequence of SEQ ID NO: 181; (28) The VH contains the amino acid sequence of SEQ ID NO: 182, and the VL contains the amino acid sequence of SEQ ID NO: 183; (29) The VH contains the amino acid sequence of SEQ ID NO: 184, and the VL contains the amino acid sequence of SEQ ID NO: 185; (30) The VH contains the amino acid sequence of SEQ ID NO: 186, and the VL contains the amino acid sequence of SEQ ID NO: 187; (31) The VH contains the amino acid sequence of SEQ ID NO: 190, and the VL contains the amino acid sequence of SEQ ID NO: 191; (32) The VH contains the amino acid sequence of SEQ ID NO: 192, and the VL contains the amino acid sequence of SEQ ID NO: 193; (33) The VH contains the amino acid sequence of SEQ ID NO: 194, and the VL contains the amino acid sequence of SEQ ID NO: 195; (34) The VH contains the amino acid sequence of SEQ ID NO: 196, and the VL contains the amino acid sequence of SEQ ID NO: 197; (35) The VH contains the amino acid sequence of SEQ ID NO: 198, and the VL contains the amino acid sequence of SEQ ID NO: 199; (36) The VH contains the amino acid sequence of SEQ ID NO: 212, and the VL contains the amino acid sequence of SEQ ID NO: 213; (37) The VH contains the amino acid sequence of SEQ ID NO: 253, and the VL contains the amino acid sequence of SEQ ID NO: 254; (38) The VH contains the amino acid sequence of SEQ ID NO: 255, and the VL contains the amino acid sequence of SEQ ID NO: 256; (39) The VH contains the amino acid sequence of SEQ ID NO: 257, and the VL contains the amino acid sequence of SEQ ID NO: 258; (40) The VH contains the amino acid sequence of SEQ ID NO: 289, and the VL contains the amino acid sequence of SEQ ID NO: 127; or (41) The VH contains the amino acid sequence of SEQ ID NO: 216, and the VL contains the amino acid sequence of SEQ ID NO:

214.

7. The isolated anti-human FD antibody construct according to any one of claims 1 to 6, wherein the anti-human FD antibody portion cross-reacts with cynomolgus factor D (cyno FD).

8. The isolated anti-human FD antibody construct according to any one of claims 1 to 7, wherein the anti-human FD antibody moiety is pH-dependent, and wherein the anti-human FD antibody moiety binds more strongly to human FD at neutral pH than it binds to human FD at acidic pH.

9. The isolated anti-human FD antibody construct according to any one of claims 1 to 8, wherein the anti-human FD antibody portion is selected from the group consisting of: full-length antibody, Fab, Fab', F(ab)2, F(ab')2, scFv, and combinations thereof.

10. The isolated anti-human FD antibody construct according to any one of claims 1 to 9, wherein the anti-human FD antibody portion is a full-length antibody (anti-human FD full-length antibody).

11. The isolated anti-human FD antibody construct of claim 10, wherein the full-length anti-human FD antibody comprises a heavy chain constant region derived from IgG4.

12. The isolated anti-human FD antibody construct of claim 11, wherein the heavy chain constant region comprises the amino acid sequence of any one of SEQ ID NO: 132-139.

13. The isolated anti-human FD antibody construct according to any one of claims 1 to 9, wherein the anti-human FD antibody portion is scFv (anti-human FD scFv).

14. The isolated anti-human FD antibody construct of claim 14, wherein the anti-human FD scFv comprises the amino acid sequence of SEQ ID NO: 219 or 220.

15. The isolated anti-human FD antibody construct according to any one of claims 1 to 14, wherein the isolated anti-human FD antibody construct further comprises a second antibody portion that specifically recognizes a component of the complement pathway.

16. The isolated anti-human FD antibody construct of claim 15, wherein the second antibody portion is selected from the group consisting of: full-length antibody, Fab, Fab', F(ab)2, F(ab')2, scFv, sdAb, and combinations thereof.

17. The isolated anti-human FD antibody construct as described in claim 15 or 16, wherein the complement pathway component is complement component 2 (C2).

18. The isolated anti-human FD antibody construct as described in claim 15 or 16, wherein the complement pathway component is complement component 5 (C5).

19. The isolated anti-human FD antibody construct according to any one of claims 15 to 18, wherein the anti-human FD antibody portion is scFv (scFv1), and wherein the second antibody portion is scFv (scFv2).

20. The isolated anti-human FD antibody construct of claim 19, wherein the isolated anti-human FD antibody construct comprises, from the N-terminus to the C-terminus: (i) scFv1 - optional connector - scFv2; (ii) scFv2 - optional connector - scFv1; (iii) scFv1 - optional connector - Fc domain - optional connector - scFv2; or (iv) scFv2 - Optional connector - Fc domain - Optional connector - scFv1.

21. The isolated anti-human FD antibody construct as described in claim 20, wherein: i) The linker contains the amino acid sequence of any one of SEQ ID NO: 221-229; and / or ii) The Fc domain contains the amino acid sequence of SEQ ID NO:

230.

22. An isolated nucleic acid, said isolated nucleic acid encoding the isolated anti-human FD antibody construct according to any one of claims 1 to 21.

23. A vector comprising the isolated nucleic acid as described in claim 22.

24. The vector of claim 23, wherein the vector is a viral vector.

25. The vector of claim 24, wherein the viral vector is an adeno-associated virus (AAV) vector or a lentiviral vector.

26. A host cell comprising the isolated nucleic acid of claim 22 or the vector of any one of claims 23 to 25.

27. A method for preparing an isolated anti-human FD antibody construct, comprising: i) Culture host cells, comprising the isolated nucleic acid of claim 22, or the vector of any one of claims 23 to 25, or the host cell of claim 26, under conditions suitable for expressing the isolated anti-human FD antibody construct; and ii) Obtain the expressed isolated anti-human FD antibody construct from the host cell.

28. A pharmaceutical composition comprising an isolated anti-human FD antibody construct according to any one of claims 1 to 21, an isolated nucleic acid according to claim 22 or a vector according to any one of claims 23 to 25, and a pharmaceutically acceptable carrier.

29. A method for treating an individual with a complement-mediated disease, comprising administering to the individual an effective amount of the pharmaceutical composition of claim 28.

30. The method of claim 29, wherein the complement-mediated disease is selected from the group consisting of: macular degeneration (MD), age-related macular degeneration (AMD), ischemia-reperfusion injury, arthritis, rheumatoid arthritis, lupus, ulcerative colitis, stroke, postoperative systemic inflammatory syndrome, asthma, allergic asthma, chronic obstructive pulmonary disease (COPD), paroxysmal nocturnal hemoglobinuria (PNH) syndrome, autoimmune hemolytic anemia (AIHA), Gaucher disease, myasthenia gravis, neuromyelitis optica (NMO), multiple sclerosis, delayed graft function, antibody-mediated rejection, atypical hemolytic uremic syndrome (aHUS), central retinal vein occlusion (CRVO), and central retinal artery occlusion (CRAO). Bullous epidermolysis, sepsis, septic shock, organ transplantation, inflammation, C3 glomerulonephropathy (C3G), membranous nephropathy, IgA nephropathy (IgAN), glomerulonephritis, thrombotic microangiopathy secondary to systemic lupus erythematosus (SLE-TMA), antineutrophil cytoplasmic antibody (ANCA) mediated vasculitis, Shiga toxin-induced HUS, antiphospholipid antibody-induced abortion, graft-versus-host disease (GvHD), bullous pemphigoid, hidradenitis suppurativa, herpetic dermatitis, Sweet's syndrome, gangrenous pyoderma, palmoplantar pustulosis and pustular psoriasis, rheumatoid neutrophilic dermatitis, subcorneal pustular dermatitis, intestinal-associated dermatitis-arthritis syndrome, neutrophilic eccrine sweat gland inflammation, linear IgA disease, and any combination thereof.

31. A method for reducing the activity of an individual's complement system, comprising administering to the individual an effective amount of the pharmaceutical composition of claim 28.

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