Complement factor H-related 4 specific antibodies and uses thereof

Abstract

The present disclosure relates to the treatment and / or prevention of age-related macular degeneration (AMD), including the advanced form of dry AMD called geographic atrophy (GA). In particular, the present disclosure provides novel therapeutic antibodies that target components of the alternative pathway of the complement activation system, including complement factor H-related (CFHR)4.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 336,751, filed April 29, 2022, which is incorporated by reference in its entirety for all purposes.

[0002] Sequence Listing The text of the computer readable sequence listing entitled "40186_601_SequenceListing", having a file size of 643,199 bytes generated on March 30, 2023, submitted herewith, is hereby incorporated by reference in its entirety.

[0003] Embodiments of the present disclosure relate to the treatment and / or prevention of age-related macular degeneration (AMD), including the advanced form of dry AMD called geographic atrophy (GA). In particular, the present disclosure provides novel therapeutic antibodies that target components of the alternative pathway of the complement activation system, including complement factor H-related (CFHR)4. [Background technology]

[0004] Age-related macular degeneration (AMD) is the leading cause of severe vision loss in adults over the age of 50. The Centers for Disease Control and Prevention estimate that 1.8 million people have AMD and an additional 7.3 million are at substantial risk of vision loss due to AMD. This eye disease occurs when there are changes in the macula, a small portion of the retina located in the inner back layer of the eye. AMD is a loss of central vision and can occur in two forms: "dry" (atrophic) and "wet" (exudative). Most people with macular degeneration have the dry form. Geographic atrophy (GA) is considered a later stage of the dry form of AMD and is characterized by a focal and progressive loss of photoreceptors. GA is less common than neovascular AMD, accounting for 10-20% of cases of legal blindness in this condition, affecting over 5 million people worldwide. There are currently no approved or effective treatments to prevent either the onset or progression of GA. However, in recent years, there have been significant advances in understanding the pathogenesis of GA, leading to several new potential treatments.

[0005] Genetic and molecular studies have identified the complement system as a key factor in the pathogenesis and progression of AMD. Many of the genetic risk variants are concentrated in genes of the alternative pathway of the complement system, and complement activation products are elevated in AMD patients. However, attempts to treat AMD via complement regulators have not yet been successful, suggesting a level of complexity that cannot be predicted from a genetic perspective alone. Therefore, new therapeutic approaches for the treatment of AMD and GA are still needed. Summary of the Invention

[0006] Embodiments of the present disclosure include antibodies against a complement factor H related 4 (CFHR4) peptide, or an antigen-binding fragment thereof. In accordance with these embodiments, the present disclosure provides a heavy chain variable region (VH) comprising complementarity determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and a light chain variable region (VL) comprising complementarity determining regions (CDRs) LCDR1, LCDR2, and LCDR3. In some embodiments, HCDR1 comprises one of the following amino acid sequences: (a) X1YX2X3X4 (SEQ ID NO: 1), where X1 is S, T, G, or N, X2 is G or Y, X3 is I or M, and X4 is S, H, or Q; (b) X1YX2X3X4 (SEQ ID NO: 21), where X1 is S, T, R, or D, X2 is T, V, A, G, S, or E, and X3 is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 29, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, , M or I, and X4 is N, S, or H; (c) X1X2X3WX4X5 (SEQ ID NO:53), where X1 is T, S, G, or I, X2 is S, R, Y, or H, X3 is D, N, H, K, or Y, X4 is W or S, and X5 is T or S; or (d) SNX1AX2WN (SEQ ID NO:88), where X1 is S, T, or N, and X2 is A or S. In some embodiments, HCDR2 comprises one of the following amino acid sequences: (a) X1IX2X3X4X5GX6TX7X8X9X 10 X 11 X 12 QX 13 (SEQ ID NO: 8), wherein X1 is W or I, X2 is S, N, or D, X3 is A or P, X4 is Y, N, or S, X5 is N, G, or S, X6 is N, S, or G, X7 is N, H, T, or S, X8 is Y or N, X9 is A or Y, and X 10 is Q or A, and X 11 is K or S, and X 12 is L or F, and X 13 is G or D, (b) X1IX2X3X4X5X6X7X8X9X 10 X 11 DX 12 VX 13X1 is S, V, Y, G or D, X2 is S or W, X3 is S, V, G, H or W, X4 is S, D, N or T, X5 is S or G, X6 is S or G, X7 is Y, S, T or R, X8 is T, K, I or V, X9 is Y, F, N or G, and X 10 is Y or H, and X 11 is A or V, and X 12 is S or P, and X 13 is R, K, or T; (c) X1X2X3X4X5GX6X7X8X9X 10 PLSX 11 X1 is E or Y, X2 is I, T, or V, X3 is Y, H, or F, X4 is H or Y, X5 is S, D, T, G, X6 is S, T, G, or N, X7 is T or I, X8 is N or K, X9 is Y, K, or S, and X 10 is N, S, K, or H, and X 11 is K or Q, or (d) X1TX2YRSX3X4X5X6X7X8X9X 10 SX 11 X 12 X (SEQ ID NO: 102), wherein X1 is R, T, M, or K, X2 is Y or F, X3 is K or R, X4 is W or L, X5 is F, Y, or S, X6 is N, S, D, or A, X7 is N, V, G, D, Y, or A, X8 is Y or F, X9 is S, A, or P, and X 10 is V, L, or A, and X 11 is V or M, and X 12 is K or S. In some embodiments, HCDR3 comprises one of the following amino acid sequences: (a) SEQ ID NOs: 15-20, (b) SEQ ID NOs: 43-52, (c) SEQ ID NOs: 77-87, or (d) SEQ ID NOs: 116-128.

[0007] According to the above embodiment, LCDR1 of the anti-CFHR4 antibody of the present disclosure comprises any of the amino acid sequences of SEQ ID NOs: 130 to 151, 198 to 199, 207 to 215, or 237 to 243, LCDR2 comprises any of the amino acid sequences of SEQ ID NOs: 153 to 174, 201 to 202, 217 to 225, or 245 to 251, and LCDR3 comprises any of the amino acid sequences of SEQ ID NOs: 175 to 196, 204 to 205, 227 to 235, or 253 to 259.

[0008] In some embodiments, the disclosure provides an antibody against a CFHR4 peptide, or an antigen-binding fragment thereof, the antibody comprising a VH comprising complementarity determining regions HCDR1, HCDR2, and HCDR3, and a VL comprising complementarity determining regions LCDR1, LCDR2, LCDR3, wherein LCDR1 comprises one of the following amino acid sequences: (a) RX1SX2X3X4X5X6X7LX8 (SEQ ID NO: 129), where X1 is A or T, X2 is Q or K, X3 is G, S, D, or N, X4 is I, F, or V, X5 is T, R, A, S, N, G, or I, and X6 is T, N, G, or I; , S, I, K, or Y, X7 is W, D, or Y, and X8 is A, T, G, N, or D; (b) RSSQX1LLHSX2GYNX3LD (SEQ ID NO: 197), where X1 is S or R, X2 is T or S, and X3 is F or Y; (c) RASQX1X2X3X4X5X6X7X8A (SEQ ID NO: 206), where X1 is S, N, or T, X2 is V or I, X3 is S or R, X4 is S, G or N, X5 is N or S, X6 is L or Y, and X7 is A, L, or V; or (d) SEQ ID NO: 236. In some embodiments, LCDR2 comprises one of the following amino acid sequences: (a) X1X2SX3LX4X5 (SEQ ID NO: 152), where X1 is G, A, T, or K, X2 is A or T, X3 is S, T, G, or N, X4 is E, Q, or L, and X5 is S, T, or G; (b) LX1SX2RAS (SEQ ID NO: 200), where X1 is A or G and X2 is N or S; (c) GASX1RAT (SEQ ID NO: 216), where X1 is T, S, or N; or (d) WASX1RES (SEQ ID NO: 244), where X1 is T, P, or N.In some embodiments, LCDR3 comprises one of the following amino acid sequences: (a) SEQ ID NOs: 175-196, (b) MQX1LQTPX2T (SEQ ID NO: 203), where X1 is A or G and X2 is Y or P, or (c) QX1YX2X3X4X5X6T (SEQ ID NO: 226), where X1 is Q or H, X2 is D or G, and X3 is N, S, or R. wherein X4 is W or S, X5 is R, P, F, Y, V, or I, and X6 is T, W, L, or I; or (d) QQX1X2X3X4PX5X6T (sequence number 252), wherein X1 is Y or F, X2 is G or Y, X3 is S or N, X4 is S, T, or I, X5 is M, Y, or R, and X6 is Y or T.

[0009] According to the above embodiments, HCDR1 of the anti-CFHR4 antibody of the present disclosure comprises any of the amino acid sequences of SEQ ID NOs: 2 to 7, 22 to 31, 54 to 64, or 89 to 101, HCDR2 comprises any of the amino acid sequences of SEQ ID NOs: 9 to 14, 33 to 42, 66 to 76, or 103 to 115, and HCDR3 comprises any of the amino acid sequences of SEQ ID NOs: 15 to 20, 43 to 52, 77 to 87, or 116 to 128.

[0010] In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:2, HCDR2 comprises the amino acid sequence of SEQ ID NO:9, and HCDR3 comprises the amino acid sequence of SEQ ID NO:15. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:3, HCDR2 comprises the amino acid sequence of SEQ ID NO:10, and HCDR3 comprises the amino acid sequence of SEQ ID NO:16. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:4, HCDR2 comprises the amino acid sequence of SEQ ID NO:11, and HCDR3 comprises the amino acid sequence of SEQ ID NO:17. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:5, HCDR2 comprises the amino acid sequence of SEQ ID NO:12, and HCDR3 comprises the amino acid sequence of SEQ ID NO:18. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:6, HCDR2 comprises the amino acid sequence of SEQ ID NO:13, and HCDR3 comprises the amino acid sequence of SEQ ID NO:19. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:7, HCDR2 comprises the amino acid sequence of SEQ ID NO:14, and HCDR3 comprises the amino acid sequence of SEQ ID NO:20. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:22, HCDR2 comprises the amino acid sequence of SEQ ID NO:33, and HCDR3 comprises the amino acid sequence of SEQ ID NO:43. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:23, HCDR2 comprises the amino acid sequence of SEQ ID NO:34, and HCDR3 comprises the amino acid sequence of SEQ ID NO:44. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:24, HCDR2 comprises the amino acid sequence of SEQ ID NO:35, and HCDR3 comprises the amino acid sequence of SEQ ID NO:45. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:25, HCDR2 comprises the amino acid sequence of SEQ ID NO:36, and HCDR3 comprises the amino acid sequence of SEQ ID NO:46. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:26, HCDR2 comprises the amino acid sequence of SEQ ID NO:37, and HCDR3 comprises the amino acid sequence of SEQ ID NO:47. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:27, HCDR2 comprises the amino acid sequence of SEQ ID NO:38, and HCDR3 comprises the amino acid sequence of SEQ ID NO:48.In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:28, HCDR2 comprises the amino acid sequence of SEQ ID NO:39, and HCDR3 comprises the amino acid sequence of SEQ ID NO:49. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:29, HCDR2 comprises the amino acid sequence of SEQ ID NO:40, and HCDR3 comprises the amino acid sequence of SEQ ID NO:50. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:30, HCDR2 comprises the amino acid sequence of SEQ ID NO:41, and HCDR3 comprises the amino acid sequence of SEQ ID NO:51. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:31, HCDR2 comprises the amino acid sequence of SEQ ID NO:42, and HCDR3 comprises the amino acid sequence of SEQ ID NO:52. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:54, HCDR2 comprises the amino acid sequence of SEQ ID NO:66, and HCDR3 comprises the amino acid sequence of SEQ ID NO:77. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:55, HCDR2 comprises the amino acid sequence of SEQ ID NO:67, and HCDR3 comprises the amino acid sequence of SEQ ID NO:78. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:56, HCDR2 comprises the amino acid sequence of SEQ ID NO:68, and HCDR3 comprises the amino acid sequence of SEQ ID NO:79. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:57, HCDR2 comprises the amino acid sequence of SEQ ID NO:69, and HCDR3 comprises the amino acid sequence of SEQ ID NO:80. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:58, HCDR2 comprises the amino acid sequence of SEQ ID NO:70, and HCDR3 comprises the amino acid sequence of SEQ ID NO:81. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:59, HCDR2 comprises the amino acid sequence of SEQ ID NO:71, and HCDR3 comprises the amino acid sequence of SEQ ID NO:82. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:60, HCDR2 comprises the amino acid sequence of SEQ ID NO:72, and HCDR3 comprises the amino acid sequence of SEQ ID NO:83.In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:61, HCDR2 comprises the amino acid sequence of SEQ ID NO:73, and HCDR3 comprises the amino acid sequence of SEQ ID NO:84. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:62, HCDR2 comprises the amino acid sequence of SEQ ID NO:74, and HCDR3 comprises the amino acid sequence of SEQ ID NO:85. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:63, HCDR2 comprises the amino acid sequence of SEQ ID NO:75, and HCDR3 comprises the amino acid sequence of SEQ ID NO:86. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:64, HCDR2 comprises the amino acid sequence of SEQ ID NO:76, and HCDR3 comprises the amino acid sequence of SEQ ID NO:87. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:89, HCDR2 comprises the amino acid sequence of SEQ ID NO:103, and HCDR3 comprises the amino acid sequence of SEQ ID NO:116. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:90, HCDR2 comprises the amino acid sequence of SEQ ID NO:104, and HCDR3 comprises the amino acid sequence of SEQ ID NO:117. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:91, HCDR2 comprises the amino acid sequence of SEQ ID NO:105, and HCDR3 comprises the amino acid sequence of SEQ ID NO:118. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:92, HCDR2 comprises the amino acid sequence of SEQ ID NO:106, and HCDR3 comprises the amino acid sequence of SEQ ID NO:119. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:93, HCDR2 comprises the amino acid sequence of SEQ ID NO:107, and HCDR3 comprises the amino acid sequence of SEQ ID NO:120. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:94, HCDR2 comprises the amino acid sequence of SEQ ID NO:108, and HCDR3 comprises the amino acid sequence of SEQ ID NO:121. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:95, HCDR2 comprises the amino acid sequence of SEQ ID NO:109, and HCDR3 comprises the amino acid sequence of SEQ ID NO:122.In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:96, HCDR2 comprises the amino acid sequence of SEQ ID NO:110, and HCDR3 comprises the amino acid sequence of SEQ ID NO:123. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:97, HCDR2 comprises the amino acid sequence of SEQ ID NO:111, and HCDR3 comprises the amino acid sequence of SEQ ID NO:124. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:98, HCDR2 comprises the amino acid sequence of SEQ ID NO:112, and HCDR3 comprises the amino acid sequence of SEQ ID NO:125. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:99, HCDR2 comprises the amino acid sequence of SEQ ID NO:113, and HCDR3 comprises the amino acid sequence of SEQ ID NO:126. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:100, HCDR2 comprises the amino acid sequence of SEQ ID NO:114, and HCDR3 comprises the amino acid sequence of SEQ ID NO:127. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 101, HCDR2 comprises the amino acid sequence of SEQ ID NO: 115, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 128. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 130, LCDR2 comprises the amino acid sequence of SEQ ID NO: 153, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 175. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 131, LCDR2 comprises the amino acid sequence of SEQ ID NO: 154, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 176. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 132, LCDR2 comprises the amino acid sequence of SEQ ID NO: 155, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 177. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 133, LCDR2 comprises the amino acid sequence of SEQ ID NO: 156, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 178. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:134, LCDR2 comprises the amino acid sequence of SEQ ID NO:157, and LCDR3 comprises the amino acid sequence of SEQ ID NO:179.In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 135, LCDR2 comprises the amino acid sequence of SEQ ID NO: 158, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 180. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 136, LCDR2 comprises the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 181. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 137, LCDR2 comprises the amino acid sequence of SEQ ID NO: 160, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 182. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 138, LCDR2 comprises the amino acid sequence of SEQ ID NO: 161, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 183. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 139, LCDR2 comprises the amino acid sequence of SEQ ID NO: 162, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 184. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 140, LCDR2 comprises the amino acid sequence of SEQ ID NO: 163, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 185. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 141, LCDR2 comprises the amino acid sequence of SEQ ID NO: 164, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 186. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 142, LCDR2 comprises the amino acid sequence of SEQ ID NO: 165, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 187. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 143, LCDR2 comprises the amino acid sequence of SEQ ID NO: 166, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 188. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 144, LCDR2 comprises the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 189. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:145, LCDR2 comprises the amino acid sequence of SEQ ID NO:168, and LCDR3 comprises the amino acid sequence of SEQ ID NO:190.In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 146, LCDR2 comprises the amino acid sequence of SEQ ID NO: 169, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 191. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 147, LCDR2 comprises the amino acid sequence of SEQ ID NO: 170, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 192. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 148, LCDR2 comprises the amino acid sequence of SEQ ID NO: 171, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 193. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 149, LCDR2 comprises the amino acid sequence of SEQ ID NO: 172, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 194. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 150, LCDR2 comprises the amino acid sequence of SEQ ID NO: 173, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 195. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 151, LCDR2 comprises the amino acid sequence of SEQ ID NO: 174, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 196. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 198, LCDR2 comprises the amino acid sequence of SEQ ID NO: 201, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 204. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 199, LCDR2 comprises the amino acid sequence of SEQ ID NO: 202, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 205. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 207, LCDR2 comprises the amino acid sequence of SEQ ID NO: 217, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 227. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:208, LCDR2 comprises the amino acid sequence of SEQ ID NO:218, and LCDR3 comprises the amino acid sequence of SEQ ID NO:228. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:209, LCDR2 comprises the amino acid sequence of SEQ ID NO:219, and LCDR3 comprises the amino acid sequence of SEQ ID NO:229. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:210, LCDR2 comprises the amino acid sequence of SEQ ID NO:220, and LCDR3 comprises the amino acid sequence of SEQ ID NO:230. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:211, LCDR2 comprises the amino acid sequence of SEQ ID NO:221, and LCDR3 comprises the amino acid sequence of SEQ ID NO:231. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:212, LCDR2 comprises the amino acid sequence of SEQ ID NO:222, and LCDR3 comprises the amino acid sequence of SEQ ID NO:232. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:213, LCDR2 comprises the amino acid sequence of SEQ ID NO:223, and LCDR3 comprises the amino acid sequence of SEQ ID NO:233.In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:214, LCDR2 comprises the amino acid sequence of SEQ ID NO:224, and LCDR3 comprises the amino acid sequence of SEQ ID NO:234. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:215, LCDR2 comprises the amino acid sequence of SEQ ID NO:225, and LCDR3 comprises the amino acid sequence of SEQ ID NO:235. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:237, LCDR2 comprises the amino acid sequence of SEQ ID NO:245, and LCDR3 comprises the amino acid sequence of SEQ ID NO:253. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:238, LCDR2 comprises the amino acid sequence of SEQ ID NO:246, and LCDR3 comprises the amino acid sequence of SEQ ID NO:254. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO:239, LCDR2 comprises the amino acid sequence of SEQ ID NO:247, and LCDR3 comprises the amino acid sequence of SEQ ID NO:255. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 240, LCDR2 comprises the amino acid sequence of SEQ ID NO: 248, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 256. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 241, LCDR2 comprises the amino acid sequence of SEQ ID NO: 249, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 257. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 242, LCDR2 comprises the amino acid sequence of SEQ ID NO: 250, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 258. In some embodiments, LCDR1 comprises the amino acid sequence of SEQ ID NO: 243, LCDR2 comprises the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259.

[0011] In some embodiments, the VH of an anti-CFHR4 antibody of the present disclosure comprises an amino acid sequence at least 90% identical to any of (a) SEQ ID NOs: 260-265, (b) SEQ ID NOs: 284-293, (c) SEQ ID NOs: 324-334, or (d) SEQ ID NOs: 368-380. In some embodiments, the VL of an anti-CFHR4 antibody of the present disclosure comprises an amino acid sequence at least 90% identical to any of (a) SEQ ID NOs: 272-277, (b) SEQ ID NOs: 304-313, (c) SEQ ID NOs: 346-356, or (d) SEQ ID NOs: 394-406. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO: 260, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 272. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO: 261, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 273. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:262, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:274. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:263, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:275. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:264, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:276. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:265, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:277. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:284, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:304. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:285, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:305. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:286, and the VL comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:306.In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:287, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:307. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:288, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:308. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:289, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:309. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:290, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:310. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:291, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:311. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:292, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:312. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:293, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:313. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:324, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:346. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:325, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:347. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:326, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:348. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:327, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:349. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:328, and the VL comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:350.In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:329, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:351. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:330, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:352. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:331, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:353. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:332, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:354. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:333, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:354. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:334, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:356. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:368, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:394. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:369, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:395. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:370, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:396. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:371, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:397. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:372, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:398. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:373, and the VL comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:399.In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:374, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:400. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:375, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:401. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:376, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:402. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:377, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:403. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:378, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:404. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:379, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:405. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:380, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:406.

[0012] In accordance with the above embodiments, the present disclosure provides anti-CFHR4 antibodies comprising various functional characteristics. In some embodiments, the anti-CFHR4 antibodies described herein bind to an antigen on CFHR4 (SEQ ID NO: 580), or a variant or isoform thereof, via interaction with an antigenic determinant (epitope) thereof. In some embodiments, binding of the anti-CFHR4 antibody to CFHR4 reduces complement activation. In some embodiments, the anti-CFHR4 antibody binds to human CFHR4b and / or binds to human CFHR4b. D It binds at approximately 100 nM or less (Figure 6).

[0013] In some embodiments, the anti-CFHR4 antibody attenuates complement component 3 (C3) convertase activity. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:330, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:352. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:293, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:313. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:378, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:404. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:377, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:403. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:376, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:402. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO: 289, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 309. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO: 371, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 397. In some embodiments, the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO: 284, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 304.

[0014] In some embodiments, the antibody cross-reacts with cynomolgus monkey CFHR4b protein (cCFHR4b). In some embodiments, the antibody cross-reacts with ATX-P-560, ATX-P-561, ATX-P-562, ATX-P-563, ATX-P-564, ATX-P-565, ATX-P-566, ATX-P-568, ATX-P-569, ATX-P-570, ATX-P-571, ATX-P-573, ATX-P-574, ATX-P-576, ATX-P-577, ATX-P-578, ATX- The antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and VL region of an antibody selected from the group consisting of ATX-P-579, ATX-P-580, ATX-P-581, ATX-P-582, ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-591, ATX-P-592, ATX-P-594, ATX-P-596, ATX-P-600, and ATX-P-604.

[0015] In some embodiments, the antibody does not cross-react with cynomolgus CFHR4b protein (cCFHR4b). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and the VL region of an antibody selected from the group consisting of ATX-P-572, ATX-P-595, ATX-P-597, ATX-P-598, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-607, ATX-P-608, ATX-P-609, and ATX-P-610.

[0016] In some embodiments, the antibody cross-reacts with complement factor H related protein 4a (CFHR4a). In some embodiments, the antibody cross-reacts with complement factor H related protein 4a (CFHR4a). In some embodiments, the antibody cross-reacts with complement factor H related protein 4a (CFHR4a). and VL regions that are at least 90% identical to the VH and VL regions of an antibody selected from the group consisting of ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-591, ATX-P-592, ATX-P-594, ATX-P-595, ATX-P-596, ATX-P-597, ATX-P-600, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-604, ATX-P-607, ATX-P-608, ATX-P-609, and ATX-P-610.

[0017] In some embodiments, the antibody does not cross-react with complement factor H related protein 4a (CFHR4a). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to ATX-P-598.

[0018] In some embodiments, the antibody cross-reacts with complement factor H related protein 3 (CFHR3). In some embodiments, the antibody cross-reacts with complement factor H related protein 3 (CFHR3). In some embodiments, the antibody cross-reacts with complement factor H related protein 3 (CFHR3). , ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-592, ATX-P-594, ATX-P-595, ATX-P-596, ATX-P-600, ATX-P-603, ATX-P-604, ATX-P-608, ATX-P-609, and ATX-P-610.

[0019] In some embodiments, the antibody does not cross-react with complement factor H related protein 3 (CFHR3). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and the VL region of an antibody selected from the group consisting of ATX-P-560, ATX-P-570, ATX-P-571, ATX-P-582, ATX-P-591, ATX-P-597, ATX-P-598, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-604, and ATX-P-607.

[0020] In some embodiments, the antibody cross-reacts with complement factor H related protein 1 (CFHL1). In some embodiments, the antibody cross-reacts with ATX-P-561, ATX-P-562, ATX-P-563, ATX-P-564, ATX-P-565, ATX-P-566, ATX-P-568, ATX-P-570, ATX-P-571, ATX-P-573, ATX-P-574, ATX-P-576, ATX-P-577, ATX-P-578, ATX-P-579, ATX- The antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and VL region of an antibody selected from the group consisting of ATX-P-580, ATX-P-581, ATX-P-582, ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-591, ATX-P-594, ATX-P-595, ATX-P-596, ATX-P-600, and ATX-P-604.

[0021] In some embodiments, the antibody does not cross-react with complement factor H related protein 1 (CFHL1). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and the VL region of an antibody selected from the group consisting of ATX-P-560, ATX-P-569, ATX-P-572, ATX-P-592, ATX-P-597, ATX-P-598, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-607, ATX-P-608, ATX-P-609, and ATX-P-610.

[0022] In accordance with the above embodiments, the anti-CFHR4 antibody of the present disclosure may be a monoclonal antibody, a human antibody, a humanized antibody, and / or a chimeric antibody. In some embodiments, the antibody is a fragment selected from the group consisting of Fab, Fab-C, Fab'-SH, Fv, scFV, and (Fab')2 fragments. In some embodiments, the anti-CFHR4 antibody is a monospecific antibody. In some embodiments, the anti-CFHR4 antibody is a bispecific antibody. In some embodiments, the anti-CFHR4 antibody comprises two or more single domain antibodies forming a bivalent, trivalent, or tetravalent antibody that recognize different epitopes on the same or different antigens.

[0023] In some embodiments, the antibody comprises a detection moiety. In some embodiments, the antibody comprises a purification moiety. In some embodiments, the antibody comprises a half-life extending moiety. In some embodiments, the half-life extending moiety comprises a polypeptide that is at least 20 amino acids in length and that comprises any combination of G, A, ST, E, and P residues. In some embodiments, the half-life extending polypeptide is attached to the C-terminus or N-terminus of the antibody.

[0024] The anti-CFHR4 antibodies of the disclosure can be administered as part of a pharmaceutical composition in a therapeutically effective amount to treat ocular diseases (e.g., AMD or GA). In some embodiments, the composition is suitable for ocular administration. In some embodiments, ocular administration comprises injection into the vitreous humor. In some embodiments, ocular administration comprises delivering the antibody using a conjunctival insert, contact lens, gel, nanoparticles, mucoadhesive polymer, ointment, solution, suspension, eye drops, and / or implant.

[0025] The embodiments of the present disclosure also include methods of treating and / or preventing AMD and / or GA. In accordance with these embodiments, the methods include administering a pharmaceutical composition comprising a therapeutically effective amount of an anti-CFHR4 antibody of the present disclosure. In some embodiments, the pharmaceutical composition is administered to the eye to treat at least one AMD symptom. In some embodiments, the AMD comprises wet AMD. In some embodiments, the AMD comprises dry AMD (e.g., GA). In some embodiments, the at least one AMD symptom comprises visual distortion, reduced central vision, blurred vision, and / or difficulty adapting to low light. In some embodiments, administering the pharmaceutical composition reduces complement activation in the subject's eye. In some embodiments, the pharmaceutical composition comprising a therapeutically effective amount of an anti-CFHR4 antibody of the present disclosure is administered at a dose ranging from about 0.0001 mg / dose to about 100 mg / dose. In some embodiments, the pharmaceutical composition is administered at a dose ranging from about 0.0001 mg / ml to about 100 mg / ml.

[0026] Embodiments of the present disclosure also include polynucleotides encoding any of the anti-CFHR4 antibodies of the present disclosure. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0027] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises (a) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:266 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:278, (b) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:267 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:279, (c) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:268 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:280, (d) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:269 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:281, (e) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:270 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:282, or (f) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:270 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:283.

[0028] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises: (a) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:294 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:314; (b) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:295 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:315; (c) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:296 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:316; (d) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:297 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:317; (e) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:298 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:318. (f) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:299 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:319; (g) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:300 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:320; (h) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:301 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:321; (i) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:302 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:322; or (j) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:303 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:323.

[0029] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises: (a) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:335 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:357; (b) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:336 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:358; (c) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:337 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:359; (d) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:338 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:360; (e) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:339 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:361; (f) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:340 (g) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:341 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:363; (h) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:342 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:364; (i) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:343 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:365; (j) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:344 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:366; or (k) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:345 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:367.

[0030] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises: (a) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:381 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:407; (b) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:382 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:408; (c) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:383 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:409; (d) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:384 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:410; (e) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:385 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:411; (f) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:386 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:412; (h) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:388 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:414; (i) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:389 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:415; (j) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:390 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:416; or (k) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:391 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:417; (l) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:392 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:418; or (m) a nucleic acid sequence that is at least 70% identical to SEQ ID NO:393 and a nucleic acid sequence that is at least 70% identical to SEQ ID NO:419.

[0031] In accordance with these embodiments, the present disclosure includes an expression vector comprising any of the polynucleotides encoding the anti-CFHR4 antibodies of the present disclosure. In some embodiments, the expression vector is suitable for producing the anti-CFHR4 antibodies of the present disclosure for delivering the antibodies to a subject. In some embodiments, the expression vector is suitable for use in gene therapy (e.g., an expression vector for delivering a polynucleotide encoding the anti-CFHR4 antibodies of the present disclosure to a subject). In some embodiments, the expression vector is an adeno-associated virus (AAV) vector or comprises an AAV backbone. In some embodiments, the expression vector is a lentivirus vector (LV) or comprises an LV backbone. In some embodiments, the expression vector is a herpes simplex virus (HSV) vector or a retrovirus vector.

[0032] In accordance with these embodiments, the disclosure also provides a method of administering ocular gene therapy to a subject in need thereof, comprising injecting a pharmaceutical composition comprising an effective amount of an expression vector described herein (e.g., an expression vector comprising a polynucleotide encoding an anti-CFHR4 antibody of the disclosure). In accordance with these embodiments, the disclosure also provides a method of treating AMD and / or GA, comprising administering a pharmaceutical composition comprising an effective amount of an expression vector described herein (e.g., an expression vector comprising a polynucleotide encoding an anti-CFHR4 antibody of the disclosure). In some embodiments, administering the pharmaceutical composition treats at least one symptom of AMD and / or GA. [Brief description of the drawings]

[0033] [Figure 1] Representative diagram of the molecular mechanism underlying the treatment of progressive age-related macular degeneration (AMD), including geographic atrophy (GA), with the anti-CFHR4 antibodies of the present disclosure. [Diagram 2]Results (see Table 1) plot odds ratios and confidence intervals for haplotype effects when the Y402 haplotype has a deletion (tagged with one of the two variants) or a pQTL variant. Associations were performed relative to a reference haplotype with OR=1. [Figure 3A] Representative ELISA results used to determine antigen-positive serum titers in CFHR4-immunized mice according to three different immunization protocols / cohorts (cohort 1). [Figure 3B] Representative ELISA results used to determine antigen-positive serum titers in CFHR4-immunized mice from three different immunization protocols / cohorts (cohort 2). [Figure 3C] Representative ELISA results used to determine antigen-positive serum titers in CFHR4-immunized mice from three different immunization protocols / cohorts (cohort 3). [Figure 4] Representative results of thermal stability of human CFHR4 monoclonal antibodies of the present disclosure (see Table 2, "ATX" antibody). Thermal stability was assessed by differential scanning fluorimetry (DSF) using a protein thermal shift (PTS) assay. [Figure 5A] Representative results of a CFHR4 antibody cross-blocking experiment, including data from a representative heatmap analyzing the ability of antibodies to block each other for binding to the antigen. [Figure 5B] Representative results of a CFHR4 antibody cross-blocking experiment, including a representative network plot that progressively groups together antibodies with similar competition profiles. [Figure 6] Representative results of CFHR4 antibody binding kinetics. [Figure 7] Representative results of CFHR4 antibody cross-reactivity with cynomolgus monkey CFHR4b protein (cCFHR4b). [Figure 8] Representative results of CFHR4 antibody cross-reactivity with human CFHR4a protein. [Figure 9]Representative results of CFHR4 antibody cross-reactivity with human CFHR3 protein. [Figure 10] Representative results of CFHR4 antibody cross-reactivity with human CFHL-1(Y402H) protein. [Figure 11A] A representative schematic of a C3 convertase assembly assay used to assess the ability of CFHR4 antibodies of the disclosure to affect C3 convertase formation. [Figure 11B] Representative dose-response curves for the top inhibitory CFHR4 antibodies are included. [Figure 11C] Figure 11B contains the calculated IC50 values ​​for each antibody tested. [Figure 11D] A representative summary of data showing at least three functional classes of anti-CFHR4 antibodies is provided. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Embodiments of the present disclosure relate to the treatment and / or prevention of age-related macular degeneration (AMD), including the advanced form of dry AMD called geographic atrophy (GA). In particular, the present disclosure provides novel therapeutic antibodies that target components of the alternative pathway of the complement activation system, including complement factor H-related (CFHR)4.

[0035] Human complement factor H-related protein (CFHR)4 belongs to the factor H family of plasma glycoproteins composed of short consensus repeat (SCR) domains. Although factor H is well known as an inhibitor of the alternative complement pathway, the function of the CFHR protein is less clear. For example, CFHR4 lacks an SCR that is homologous to the complement inhibitory domain of factor H. However, its complement regulatory activity is not fully understood. Previous studies have shown that CFHR4 binds C-reactive protein through its most N-terminal SCR, which leads to activation of the classical complement pathway. CFHR4 binds C3b through its C-terminus, but the importance of this interaction is not fully understood. Recent reports have shown that CFHR4 may serve as a platform for the assembly of the alternative pathway C3 convertase by binding to C3b. This is based, at least in part, on the sustained ability of CFHR4-bound C3b to bind factor B and properdin, resulting in an active convertase that generates C3a and C3b from C3. CFHR4-C3bBb convertase is less susceptible to factor H-mediated decay compared to C3bBb convertase. CFHR4 mutants containing exchanges of conserved residues within the C-terminal C3b-binding site showed significantly reduced C3b binding and alternative pathway complement activation. These initial data suggest that CFHR4 acts as an enhancer of opsonization by promoting complement activation, in contrast to the complement inhibitor factor H.

[0036] Complement is a powerful effector system of innate immunity, playing a major role in the elimination of microorganisms, inflammatory processes, clearance of cellular debris, and regulation of adaptive immunity. Multilevel regulation ensures complement activation on dangerous surfaces but prevents harmful effects on host cells and tissues. This is achieved by the expression of complement inhibitors on host cell surfaces and their absence on foreign or altered self surfaces, as well as by soluble regulators with distinct binding specificities for different surfaces. Plasma glycoprotein factor H (FH) is the major soluble inhibitor of the alternative complement pathway (AP). It prevents the formation of the AP C3 convertase C3bBb by blocking the binding of factor B (FB) to C3b and accelerates the decay of existing C3 convertase by displacing Bb. It also acts as a cofactor for the serine protease factor I (FI) in the cleavage of C3b to inactive C3b (iC3b), which can no longer form the convertase. Through these mechanisms, FH inhibits the amplification of the complement cascade both in the fluid phase and on the surface of host cells. FH, its splicing variant complement factor H-like protein 1 (CFHL1), and five complement factor H-related proteins (CFHR1-CFHR5) encoded by separate genes constitute the human factor H protein family. FH is composed of 20 short consensus repeat (SCR) domains, CFHL1 contains SCRs 1-7 of FH, and CFHR proteins consist of 4-9 SCRs that are homologous to various domains of FH. Although CFHL1 shares the complement inhibitory activity of FH, the physiological roles of CFHR proteins are poorly understood.

[0037] CFHR4 is detected as two distinct glycoproteins in human plasma. The 86 kDa long isoform, termed CFHR4A, consists of nine SCRs. The shorter isoform of approximately 45 kDa, termed CFHR4B, is composed of five SCRs, representing SCR1 and SCR6-9 of CFHR4A. In CFHR4A, SCR1-4 and SCR5-8 are highly related based on nucleotide and amino acid sequence identity, which is likely the result of intramolecular duplication. Like all CFHRs, both CFHR4 isoforms lack SCRs homologous to FH and the N-terminal complement inhibitory domains SCR1-4 of CFHL1. The two most C-terminal domains of CFHR4A and CFHR4B are homologous to the C-terminal FH domains SCR19-20, which contain the C3b / C3d binding sites. CFHR4B has been shown to bind to the C3d region of C3b via its C-terminal SCR4-5. However, no significant function is associated with this C3b-binding capacity, except for a slight enhancement of the cofactor activity of FH in the presence of high CFHR4B concentrations. The CFHR4A isoform has not yet been characterized for its interaction with C3b and complement regulatory activity.

[0038] Age-related macular degeneration (AMD) is a progressive retinal disease, the early stages of which are characterized by a relative paucity of small drusen in the macula. As AMD progresses, the size and number of drusen increase, eventually leading to more advanced stages of AMD. Two forms of progressive AMD are distinguished. The first form, neovascular AMD, is characterized by the infiltration of abnormal blood vessels into the retina. These newly formed blood vessels are vulnerable, and when they break, they leak retinal blood components, leading to sudden vision loss. The second form of progressive AMD, geographic atrophy (GA), is the result of a gradual degeneration of the RPE and photoreceptor cells. Although neovascularization occurs in only 15-20% of AMD cases, it accounts for the majority of vision loss caused by AMD. Drugs targeting vascular endothelial growth factor (VEGF), one of the central molecules in angiogenesis, have proven to be highly successful in neovascular AMD. However, there is no available treatment for the majority of the remaining cases of early, intermediate, or geographic atrophic AMD, and furthermore, there is no effective means of preventing progression from early to advanced stages. Although AMD is known to be the result of a complex interaction of environmental and genetic risk factors, studies on the molecular constituents of drusen suggest that AMD may have an immunological component. This suggestion arises from the discovery within drusen of proteins involved in inflammatory and / or other immune-related responses, including components of the complement system.

[0039] With this in mind, experiments were conducted to determine the role CFHR4 may play in the pathogenesis of AMD and GA and concomitantly develop a therapeutic platform based on modulation of CFHR4 activity using anti-CFHR4 antibodies.

[0040] definition To facilitate understanding of the present technology, several terms and phrases are defined below. Additional definitions are set forth throughout the detailed description.

[0041] In the context of the embodiments of the present disclosure (particularly in view of the claims that follow), the use of the terms "a," "an," and "the," as well as "at least one" and similar referents should be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term "at least one" (e.g., "at least one of A and B") following a list of one or more items should be construed to mean one item selected from the listed items (A or B), or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms "comprising," "having," "including," and "containing" should be construed as open-ended terms (i.e., meaning "including but not limited to"), unless otherwise noted. The recitation of ranges of values ​​herein is intended to serve merely as a shorthand way of referring individually to each individual value falling within the range, and each individual value is incorporated herein as if it were individually recited herein, unless otherwise indicated herein. All methods described herein may be performed in any suitable order, unless otherwise indicated herein, unless otherwise clearly contradicted by context. The use of some and all examples or exemplary language (e.g., "e.g.") provided herein is intended merely to better illustrate various embodiments of the disclosure, and does not limit the scope of these embodiments, unless otherwise asserted. No language in this specification should be construed as indicating any non-claimed element as essential to the practice of various embodiments of the disclosure.

[0042] Additionally, as used herein, the term "or" is an inclusive "or" operator and is equivalent to the term "and / or" unless the context clearly dictates otherwise. The term "based on" is not exclusive and allows for based on additional unlisted factors unless the context clearly dictates otherwise. Additionally, throughout this specification, the meanings of "a," "an," and "the" include plural referents. The meaning of "in" includes "in" and "on."

[0043] The transitional phrase "consisting essentially of," when used in the claims of this application, limits the scope of the claim to certain substances or steps "and which do not materially affect the basic and novel characteristic(s)" of the claimed invention, as stated in In re Herz, 537 F.2d 549,551-52,190 USPQ 461,463 (CCPA 1976). For example, a composition "consisting essentially of" may contain unrecited contaminants, although the recited elements are present, at levels that do not alter the function of the recited composition compared to a pure composition, i.e., a composition "consisting essentially of" the recited components.

[0044] The term "one or more" as used herein refers to a number greater than 1. For example, the term "one or more" encompasses any of the following: 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 20 or more, 50 or more, 100 or more, or even more.

[0045] "One or more, but less than a greater number," "Two or more, but less than a greater number," "Three or more, but less than a greater number," "Four or more, but less than a greater number," "Five or more, but less than a greater number," "Six or more, but less than a greater number," "Seven or more, but less than a greater number," "Eight or more, but less than a greater number," "Nine or more, but less than a greater number," "Ten or more, but less than a greater number," "Eleven or more, but less than a greater number," "Twelve or more, but less than a greater number," "Thirteen or more, but less than a greater number," "Fourteen or more, but less than a greater number," or "Fifteen or more, but less than a greater number" are not limited to the higher number. For example, the higher number can be 10,000, 1,000, 100, 50, etc. For example, the larger number can be comprised by about 50 (e.g., 50, 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 32, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2).

[0046] The term "immunoglobulin" or "antibody" as used herein refers to a protein found in the blood or other body fluids of vertebrates and is used by the immune system to recognize and neutralize foreign substances such as bacteria and viruses. Typically, an immunoglobulin or antibody is a protein that contains at least one complementarity determining region (CDR). The CDRs form the "hypervariable region" of the antibody, which is responsible for antigen binding (explained further below). A whole antibody is typically composed of four polypeptides: two identical copies of a heavy (H) chain polypeptide and two identical copies of a light (L) chain polypeptide. Each of the heavy chains contains one N-terminal variable (V H ) region and three C-terminal constant (C H1 , C H2 , and C H3 ) region, and each light chain contains one N-terminal variable (V L ) region and one C-terminal constant (C L) region. The light chains of antibodies can be assigned to one of two different types, either kappa (κ) or lambda (λ), based on the amino acid sequence of their constant domain. In a typical antibody, each light chain is linked to a heavy chain by a disulfide bond, and the two heavy chains are linked to each other by disulfide bonds. The variable region of the light chain is aligned with the variable region of the heavy chain, and the constant region of the light chain is aligned with the first constant region of the heavy chain. The remaining constant regions of the heavy chains are aligned with each other.

[0047] The variable regions of each pair of light and heavy chains form the antigen-binding site of an antibody. H and V L The regions have the same general structure, with each region containing four framework (FW or FR) regions. As used herein, the term "framework region" refers to a relatively conserved amino acid sequence in the variable region located between the CDRs. There are four framework regions in each variable domain, which are called FR1, FR2, FR3, and FR4. The framework regions form a β-sheet that provides the structural framework of the variable region (see, for example, CA Janeway et al. (eds.), Immunobiology, 5th Ed., Garland Publishing, New York, NY (2001)).

[0048] The framework regions are connected by three CDRs. As discussed above, the three CDRs, known as CDR1, CDR2, and CDR3, form the "hypervariable region" of the antibody, which is involved in antigen binding. The CDRs form loops that connect, and in some cases are part of, the beta-sheet structure formed by the framework regions. The constant regions of the light and heavy chains are not directly involved in binding the antibody to the antigen, although the constant regions can affect the orientation of the variable regions. The constant regions also exhibit various effector functions, such as participating in antibody-dependent complement-mediated lysis or antibody-dependent cellular cytotoxicity through interactions with effector molecules and cells.

[0049] As used herein, when an antibody or other entity (e.g., an antigen-binding domain) "specifically recognizes" or "specifically binds" to an antigen or epitope, it preferentially recognizes the antigen in a complex mixture of proteins and / or macromolecules and binds to the antigen or epitope with substantially higher affinity than it does to other entities that do not display the antigen or epitope. In this regard, "substantially higher affinity" means an affinity that is high enough to allow detection of the antigen or epitope as distinguished from the entity using a desired assay or measurement device. Typically, it is at least 10 7 M -1 (For example, >10 7 M -1 , >10 8 M -1 , >10 9 M -1 , >10 10 M -1 , >10 11 M -1 , >10 12 M -1 , >10 13 M -1 etc.) binding constant (K a ) means a binding affinity having a specific epitope. In certain such embodiments, the antibody can bind to different antigens as long as the different antigens contain that particular epitope. In certain cases, for example, homologous proteins from different species may contain the same epitope.

[0050] The terms "antibody fragment," "antibody fragment," and "antigen-binding fragment" of an antibody are used interchangeably herein and refer to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (see generally Holliger et al., Nat. Biotech., 23(9):1126-1129 (2005)). Any antigen-binding fragment of an antibody described herein is within the scope of the present disclosure. An antibody fragment desirably contains, for example, one or more CDRs, a variable region (or a portion thereof), a constant region (or a portion thereof), or a combination thereof. Examples of antibody fragments include: (i) a V L , V H , CL , and C H1 (ii) a F(ab')2 fragment, which is a bivalent fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; and (iii) a V(ab')2 fragment, which is a monovalent fragment consisting of a V domain of a single arm of an antibody. L and V H (iv) Fab' fragments resulting from cleavage of the disulfide bridges of the F(ab')2 fragment using mild reducing conditions; (v) disulfide-stabilized Fv fragments (dsFv); and (vi) single chain variable region domains of an antibody that specifically binds to an antigen (V H or V L These include, but are not limited to, domain antibodies (dAbs), which are polypeptides.

[0051] The term "monoclonal antibody" as used herein refers to an antibody produced by a single clone of B lymphocytes against a single epitope on an antigen. Monoclonal antibodies are typically produced using hybridoma technology as first described in Kohler and Milstein, Eur. J. Immunol., 5:511-519 (1976). Monoclonal antibodies can also be produced using recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), isolated from phage display antibody libraries (see, e.g., Clackson et al. Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991)), or produced from transgenic mice carrying a fully human immunoglobulin system (see, e.g., Lonberg, Nat. Biotechnol., 23(9):1117-25 (2005), and Lonberg, Handb. Exp. Pharmacol., 181:69-97 (2008)). In contrast, "polyclonal" antibodies are antibodies secreted by various B cell lineages within an animal. Polyclonal antibodies are a population of immunoglobulin molecules that recognize multiple epitopes on the same antigen.

[0052] As used herein, the terms "nucleic acid", "polynucleotide", "nucleotide sequence" and "oligonucleotide" are used interchangeably and refer to polymers or oligomers of pyrimidine and / or purine bases, preferably cytosine, thymine and uracil, and adenine and guanine, respectively (see Albert L. Lehninger, Principles of Biochemistry, at 793-800 (Worth Pub. 1982)). These terms encompass any deoxyribonucleotide, ribonucleotide or peptide nucleic acid building blocks, and any chemical variants thereof, such as methylated, hydroxymethylated or glycosylated forms of these bases. The polymers or oligomers may be heterogeneous or homogeneous in composition and may be isolated from naturally occurring sources or may be artificially or synthetically produced. Furthermore, the nucleic acid may be DNA or RNA, or a mixture thereof, and may exist permanently or transiently in single-stranded or double-stranded forms, including homoduplexes, heteroduplexes and hybrid states. In some embodiments, a nucleic acid or nucleic acid sequence includes other types of nucleic acid structures, such as, for example, a DNA / RNA helix, a peptide nucleic acid (PNA), a morpholino nucleic acid (see, e.g., Braasch and Corey, Biochemistry, 41(14):4503-4510 (2002), and U.S. Pat. No. 5,034,506), a locked nucleic acid (LNA, see Wahlestedt et al., Proc. Natl. Acad. Sci. USA, 97:5633-5638 (2000)), a cyclohexenyl nucleic acid (see Wang, J. Am. Chem. Soc., 122:8595-8602 (2000)), and / or a ribozyme. The terms "nucleic acid" and "nucleic acid sequence" can also encompass strands that include non-natural nucleotides, modified nucleotides, and / or non-nucleotide components (e.g., "nucleotide analogs") that can exhibit the same function as natural nucleotides.

[0053] The terms "peptide," "polypeptide," and "protein" are used interchangeably herein to refer to polymeric forms of amino acids of any length, which can include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones.

[0054] As used herein, "nucleic acid" or "nucleic acid molecule" generally refers to any ribonucleic acid or deoxyribonucleic acid, which may be unmodified or modified DNA or RNA. "Nucleic acid" includes, but is not limited to, single-stranded and double-stranded nucleic acids. As used herein, the term "nucleic acid" also includes DNA as described above that contains one or more modified bases. Thus, DNA with backbone modifications for stability or other reasons is a "nucleic acid". As used herein, the term "nucleic acid" encompasses chemically, enzymatically, or metabolically modified forms of nucleic acid, as well as the DNA of, for example, viruses and cells in their characteristic chemical forms, including simple and complex cells.

[0055] The term "oligonucleotide" or "polynucleotide" or "nucleotide" or "nucleic acid" refers to a molecule that contains two or more deoxyribonucleotides or ribonucleotides, preferably more than three, and usually more than ten. The exact size will depend on many factors and depends on the ultimate function or use of the oligonucleotide. Oligonucleotides can be produced in any manner, including chemical synthesis, DNA replication, reverse transcription, or a combination thereof. Typical deoxyribonucleotides of DNA are thymine, adenine, cytosine, and guanine. Typical ribonucleotides of RNA are uracil, adenine, cytosine, and guanine.

[0056] The terms "complementary" and "complementarity" refer to nucleotides (e.g., a single nucleotide) or polynucleotides (e.g., a sequence of nucleotides) related by the base-pairing rules. For example, the sequence 5'-AGT-3' is complementary to the sequence 3'-TCA-5'. Complementarity can be "partial," in which only a portion of the nucleic acid bases match according to the base-pairing rules. Alternatively, there can be "complete" or "total" complementarity between nucleic acids. The degree of complementarity between strands of nucleic acids affects the efficiency and strength of hybridization between strands of nucleic acids. This is particularly important in amplification reactions and in detection methods that depend on binding between nucleic acids.

[0057] The term "gene" refers to a nucleic acid (e.g., DNA or RNA) sequence that comprises coding sequences necessary for the production of an RNA or a polypeptide or its precursor. A functional polypeptide can be encoded by a full-length coding sequence or by any portion of a coding sequence, so long as the desired activity or functional property of the polypeptide (e.g., enzymatic activity, ligand binding, signal transduction, etc.) is retained. When used in reference to a gene, the term "portion" refers to fragments of that gene. These fragments can range in size from a few nucleotides to the entire gene sequence minus one nucleotide. Thus, "nucleotides comprising at least a portion of a gene" can include fragments of a gene or the entire gene.

[0058] The term "gene" also includes the coding region of a structural gene and includes sequences (e.g., including coding, regulatory, structural and other sequences) located adjacent to the coding region at both the 5' and 3' ends, at a distance of about 1 kb on either end, such that the gene corresponds to the length of the full-length mRNA. Sequences that are 5' of the coding region and present on the mRNA are referred to as 5' non-translated or untranslated sequences. Sequences that are 3' or downstream of the coding region and present on the mRNA are referred to as 3' non-translated or untranslated sequences. The term "gene" encompasses both cDNA and genomic forms of a gene. In some organisms (e.g., eukaryotes), genomic forms or clones of a gene contain the coding region interrupted by non-coding sequences referred to as either "introns," "intervening regions," or "intervening sequences." Introns are segments of a gene that are transcribed into nuclear RNA (hnRNA) and may contain regulatory elements such as enhancers. Introns are removed or "spliced ​​out" from the nuclear or primary transcript and are therefore absent in the messenger RNA (mRNA) transcript, which functions during translation to specify the sequence or order of amino acids in a nascent polypeptide.

[0059] In addition to containing introns, genomic forms of a gene may also include sequences located both 5' and 3' to the sequences present in the RNA transcript. These sequences are referred to as "flanking" sequences or regions (these flanking sequences are located 5' or 3' to the untranslated sequences present in the mRNA transcript). The 5' flanking region may contain regulatory sequences such as promoters and enhancers that control or influence the transcription of the gene. The 3' flanking region may contain sequences that direct the termination of transcription, post-transcriptional cleavage, and polyadenylation.

[0060] When referring to a gene, the term "wild type" refers to a gene that has the characteristics of a gene isolated from a naturally occurring source. When referring to a gene product, the term "wild type" refers to a gene product that has the characteristics of a gene product isolated from a naturally occurring source. When referring to a protein, the term "wild type" refers to a protein that has the characteristics of a naturally occurring protein. When used with respect to an object, the term "naturally occurring" refers to the fact that the object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including a virus) that can be isolated from a natural source and has not been intentionally modified by the hand of man in a laboratory is naturally occurring. A wild type gene is often that gene or allele that is most frequently observed in a population and is therefore arbitrarily referred to as the "normal" or "wild type" form of the gene. In contrast, when referring to a gene or gene product, the term "modified" or "mutated" refers to a gene or gene product, respectively, that exhibits modifications in sequence and / or functional properties (i.e., altered characteristics) when compared to a wild type gene or gene product. It is noted that naturally occurring mutants can be isolated, which are identified by the fact that they have altered characteristics when compared to the wild-type gene or gene product.

[0061] The term "allele" refers to genetic variations, including, but not limited to, variants and mutations, polymorphic loci, and single nucleotide polymorphic loci, frameshifts, and splice variants. An allele can occur naturally in a population, or it can arise during the lifetime of any particular individual in this population.

[0062] Thus, when used in reference to a nucleotide sequence, the terms "variant" and "mutant" refer to a nucleic acid sequence that differs by one or more nucleotides from another, usually related, nucleotide sequence. A "variant" is the difference between two different nucleotide sequences, generally one sequence being a reference sequence.

[0063] The terms "immunogen" and "antigen" are used interchangeably herein and refer to any molecule, compound, or substance that induces an immune response in an animal (e.g., a mammal). An "immune response" may involve, for example, antibody production and / or activation of immune effector cells. An antigen in the context of this disclosure may include any subunit, fragment, or epitope of any proteinaceous or non-proteinaceous (e.g., carbohydrate or lipid) molecule that elicits an immune response in a mammal. The term "epitope" refers to the sequence of an antigen that is recognized by an antibody or an antigen receptor. Epitopes are also referred to in the art as "antigenic determinants." In certain embodiments, an epitope is a region of an antigen that is specifically bound by an antibody. In certain embodiments, an epitope may include groupings of chemically active surfaces of a molecule, such as amino acids, sugar side chains, phosphoryl, or sulfonyl groups. In certain embodiments, an epitope may have specific three-dimensional structural characteristics (e.g., a "conformational" epitope) and / or specific charge characteristics. The antigen may be a protein or peptide of viral, bacterial, parasitic, fungal, protozoan, prion, cellular, or extracellular origin, which elicits an immune response in a mammal, preferably leading to protective immunity.

[0064] As used herein, a "pharmaceutically acceptable carrier" generally refers to an ingredient in a pharmaceutical formulation that is non-toxic to a subject other than the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0065] As used herein, the term "pharmaceutical formulation" generally refers to a preparation that is in a form that allows the biological activity of the active ingredient contained therein (e.g., an anti-CFHR4 antibody, antibody conjugate, fusion protein, or polymer formulation) to be effective, and that does not contain additional components that are unacceptably toxic to the subject to which the formulation is administered.

[0066] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") generally refers to a clinical intervention aimed at altering the natural course of the individual being treated, and can be performed for prophylaxis or during the clinical pathology process. Desirable effects of treatment include, but are not limited to, prevention of disease onset or recurrence, relief of symptoms, reduction of any direct or indirect pathological consequences of the disease, prevention of metastasis, reduction in the rate of disease progression, amelioration or alleviation of disease symptoms, and remission or improvement of prognosis. In some embodiments, the anti-CFHR4 antibodies of the present disclosure, or other compositions comprising the anti-CFHR4 antibodies of the present disclosure (e.g., antibody conjugates, fusion proteins, or polymer formulations) are used to delay the onset of disease or to slow the progression of disease.

[0067] The term "half-life" as used herein generally refers to the time required for the concentration of a substance (e.g., an anti-CFHR4 antibody, antibody conjugate, fusion protein (e.g., a Fab fusion protein), or polymer formulation) to decrease by half in vivo (e.g., in the eye (e.g., the vitreous)) or in vitro.

[0068] As used herein, an "effective amount" of an agent, e.g., a pharmaceutical formulation, generally refers to an amount effective to achieve the desired therapeutic or prophylactic result, at dosages and for periods of time necessary to achieve same.

[0069] An "individual" or "subject" is a mammal. Mammals include, but are not limited to, domestic animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain embodiments, an individual or subject is a human. A "subject" can be a "patient."

[0070] Anti-CFHR4 antibody As further described herein, embodiments of the present disclosure relate to the treatment and / or prevention of age-related macular degeneration (AMD), including the advanced form of dry AMD called geographic atrophy (GA). In particular, the present disclosure provides novel therapeutic antibodies that target components of the alternative pathway of the complement activation system, including complement factor H-related (CFHR)4.

[0071] As further described herein, anti-CFHR4 antibodies have been generated and their structural and functional properties described. Based on these data, embodiments of the disclosure include anti-CFHR4 antibodies, or antigen-binding fragments thereof, that include a heavy chain variable region (VH) that includes complementarity determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and a light chain variable region (VL) that includes complementarity determining regions (CDRs) LCDR1, LCDR2, and LCDR3. In some embodiments, HCDR1 comprises one of the following amino acid sequences: (a) X1YX2X3X4 (SEQ ID NO: 1), where X1 is S, T, G, or N, X2 is G or Y, X3 is I or M, and X4 is S, H, or Q; (b) X1YX2X3X4 (SEQ ID NO: 21), where X1 is S, T, R, or D, X2 is T, V, A, G, S, or E, and X3 is , M or I, and X4 is N, S, or H; (c) X1X2X3WX4X5 (SEQ ID NO:53), where X1 is T, S, G, or I, X2 is S, R, Y, or H, X3 is D, N, H, K, or Y, X4 is W or S, and X5 is T or S; or (d) SNX1AX2WN (SEQ ID NO:88), where X1 is S, T, or N, and X2 is A or S. In some embodiments, HCDR2 comprises one of the following amino acid sequences: (a) X1IX2X3X4X5GX6TX7X8X9X 10 X 11 X 12 QX 13(SEQ ID NO: 8), wherein X1 is W or I, X2 is S, N, or D, X3 is A or P, X4 is Y, N, or S, X5 is N, G, or S, X6 is N, S, or G, X7 is N, H, T, or S, X8 is Y or N, X9 is A or Y, and X 10 is Q or A, and X 11 is K or S, and X 12 is L or F, and X 13 is G or D, (b) X1IX2X3X4X5X6X7X8X9X 10 X 11 DX 12 VX 13 X1 is S, V, Y, G or D, X2 is S or W, X3 is S, V, G, H or W, X4 is S, D, N or T, X5 is S or G, X6 is S or G, X7 is Y, S, T or R, X8 is T, K, I or V, X9 is Y, F, N or G, and X 10 is Y or H, and X 11 is A or V, and X 12 is S or P, and X 13 is R, K, or T; (c) X1X2X3X4X5GX6X7X8X9X 10 PLSX 11 X1 is E or Y, X2 is I, T, or V, X3 is Y, H, or F, X4 is H or Y, X5 is S, D, T, G, X6 is S, T, G, or N, X7 is T or I, X8 is N or K, X9 is Y, K, or S, and X 10 is N, S, K, or H, and X 11 is K or Q, or (d) X1TX2YRSX3X4X5X6X7X8X9X 10 SX 11 X 12X (SEQ ID NO: 102), wherein X1 is R, T, M, or K, X2 is Y or F, X3 is K or R, X4 is W or L, X5 is F, Y, or S, X6 is N, S, D, or A, X7 is N, V, G, D, Y, or A, X8 is Y or F, X9 is S, A, or P, and X 10 is V, L, or A, and X 11 is V or M, and X 12 is K or S. In some embodiments, HCDR3 comprises one of the following amino acid sequences: (a) SEQ ID NOs: 15-20, (b) SEQ ID NOs: 43-52, (c) SEQ ID NOs: 77-87, or (d) SEQ ID NOs: 116-128.

[0072] In addition to the above HCDR1, HCDR2, and HCDR3 sequences, the anti-CFHR4 antibody of the present disclosure comprises an LCDR1 comprising the amino acid sequence of any of SEQ ID NOs: 130 to 151, 198 to 199, 207 to 215, or 237 to 243; an LCDR2 comprising the amino acid sequence of any of SEQ ID NOs: 153 to 174, 201 to 202, 217 to 225, or 245 to 251; and an LCDR3 comprising the amino acid sequence of any of SEQ ID NOs: 175 to 196, 204 to 205, 227 to 235, or 253 to 259.

[0073] In some embodiments, the disclosure provides an anti-CFHR4 antibody, or antigen-binding fragment thereof, comprising a VH comprising complementarity determining regions HCDR1, HCDR2, and HCDR3, and a VL comprising complementarity determining regions LCDR1, LCDR2, and LCDR3. In some embodiments, LCDR1 has the following amino acid sequence: (a) RX1SX2X3X4X5X6X7LX8 (SEQ ID NO: 129), where X1 is A or T, X2 is Q or K, X3 is G, S, D, or N, X4 is I, F, or V, X5 is T, R, A, S, N, G, or I, X6 is T, N, G, S, I, K, or Y, X7 is W, D, or Y, and X8 is A, T, G, N, or D; and (b) RSSQX1L LHSX2GYNX3LD (SEQ ID NO: 197), wherein X1 is S or R, X2 is T or S, and X3 is F or Y; (c) RASQX1X2X3X4X5X6X7X8A (SEQ ID NO: 206), wherein X1 is S, N, or T, X2 is V or I, X3 is S or R, X4 is S, G or N, X5 is N or S, X6 is L or Y, and X7 is A, L, or V; or (d) comprising one of SEQ ID NO: 236. In some embodiments, LCDR2 comprises one of the following amino acid sequences: (a) X1X2SX3LX4X5 (SEQ ID NO: 152), where X1 is G, A, T, or K, X2 is A or T, X3 is S, T, G, or N, X4 is E, Q, or L, and X5 is S, T, or G; (b) LX1SX2RAS (SEQ ID NO: 200), where X1 is A or G and X2 is N or S; (c) GASX1RAT (SEQ ID NO: 216), where X1 is T, S, or N; or (d) WASX1RES (SEQ ID NO: 244), where X1 is T, P, or N.In some embodiments, LCDR3 comprises one of the following amino acid sequences: (a) SEQ ID NOs: 175-196, (b) MQX1LQTPX2T (SEQ ID NO: 203), where X1 is A or G and X2 is Y or P, or (c) QX1YX2X3X4X5X6T (SEQ ID NO: 226), where X1 is Q or H, X2 is D or G, and X3 is N, S, or R. wherein X4 is W or S, X5 is R, P, F, Y, V, or I, and X6 is T, W, L, or I; or (d) QQX1X2X3X4PX5X6T (sequence number 252), wherein X1 is Y or F, X2 is G or Y, X3 is S or N, X4 is S, T, or I, X5 is M, Y, or R, and X6 is Y or T.

[0074] In addition to the above LCDR1, LCDR2, and LCDR3 sequences, the anti-CFHR4 antibody of the present disclosure comprises an HCDR1 comprising any of the amino acid sequences of SEQ ID NOs: 2 to 7, 22 to 31, 54 to 64, or 89 to 101; an HCDR2 comprising any of the amino acid sequences of SEQ ID NOs: 9 to 14, 33 to 42, 66 to 76, or 103 to 115; and an HCDR3 comprising any of the amino acid sequences of SEQ ID NOs: 15 to 20, 43 to 52, 77 to 87, or 116 to 128.

[0075] In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:2, an HCDR2 of SEQ ID NO:9, and an HCDR3 of SEQ ID NO:15. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:3, an HCDR2 of SEQ ID NO:10, and an HCDR3 of SEQ ID NO:16. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:4, an HCDR2 of SEQ ID NO:11, and an HCDR3 of SEQ ID NO:17. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:5, an HCDR2 of SEQ ID NO:12, and an HCDR3 of SEQ ID NO:18. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:6, an HCDR2 of SEQ ID NO:13, and an HCDR3 of SEQ ID NO:19. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:7, HCDR2 comprises the amino acid sequence of SEQ ID NO:14, and an HCDR3 of SEQ ID NO:20. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:22, an HCDR2 of SEQ ID NO:33, and an HCDR3 of SEQ ID NO:43. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:23, an HCDR2 of SEQ ID NO:34, and an HCDR3 of SEQ ID NO:44. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:24, an HCDR2 of SEQ ID NO:35, and an HCDR3 of SEQ ID NO:45. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:25, an HCDR2 of SEQ ID NO:36, and an HCDR3 of SEQ ID NO:46. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:26, an HCDR2 of SEQ ID NO:37, and an HCDR3 of SEQ ID NO:47. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:27, an HCDR2 of SEQ ID NO:38, and an HCDR3 of SEQ ID NO:48. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 28, an HCDR2 of SEQ ID NO: 39, and an HCDR3 of SEQ ID NO: 49. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 29, an HCDR2 of SEQ ID NO: 40, and an HCDR3 of SEQ ID NO: 50.In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:30, an HCDR2 of SEQ ID NO:41, and an HCDR3 of SEQ ID NO:51. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:31, an HCDR2 of SEQ ID NO:42, and an HCDR3 of SEQ ID NO:52. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:54, an HCDR2 of SEQ ID NO:66, and an HCDR3 of SEQ ID NO:77. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:55, an HCDR2 of SEQ ID NO:67, and an HCDR3 of SEQ ID NO:78. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:56, an HCDR2 of SEQ ID NO:68, and an HCDR3 of SEQ ID NO:79. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:57, an HCDR2 of SEQ ID NO:69, and an HCDR3 of SEQ ID NO:80. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:58, an HCDR2 of SEQ ID NO:70, and an HCDR3 of SEQ ID NO:81. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:59, an HCDR2 of SEQ ID NO:71, and an HCDR3 of SEQ ID NO:82. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:60, an HCDR2 of SEQ ID NO:72, and an HCDR3 of SEQ ID NO:83. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:61, an HCDR2 of SEQ ID NO:73, and an HCDR3 of SEQ ID NO:84. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:62, an HCDR2 of SEQ ID NO:74, and an HCDR3 of SEQ ID NO:85. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:63, an HCDR2 of SEQ ID NO:75, and an HCDR3 of SEQ ID NO:86. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 64, an HCDR2 of SEQ ID NO: 76, and an HCDR3 of SEQ ID NO: 87. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 89, an HCDR2 of SEQ ID NO: 103, and an HCDR3 of SEQ ID NO: 116. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 90, an HCDR2 of SEQ ID NO: 104, and an HCDR3 of SEQ ID NO: 117.In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:91, an HCDR2 of SEQ ID NO:105, and an HCDR3 of SEQ ID NO:118. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:92, an HCDR2 of SEQ ID NO:106, and an HCDR3 of SEQ ID NO:119. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:93, an HCDR2 of SEQ ID NO:107, and an HCDR3 of SEQ ID NO:120. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:94, an HCDR2 of SEQ ID NO:108, and an HCDR3 of SEQ ID NO:121. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:95, an HCDR2 of SEQ ID NO:109, and an HCDR3 of SEQ ID NO:122. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO:96, an HCDR2 of SEQ ID NO:110, and an HCDR3 of SEQ ID NO:123. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 97, an HCDR2 of SEQ ID NO: 111, and an HCDR3 of SEQ ID NO: 124. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 98, an HCDR2 of SEQ ID NO: 112, and an HCDR3 of SEQ ID NO: 125. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 99, an HCDR2 of SEQ ID NO: 113, and an HCDR3 of SEQ ID NO: 126. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 100, an HCDR2 of SEQ ID NO: 114, and an HCDR3 of SEQ ID NO: 127. In some embodiments, the anti-CFHR4 antibody comprises an HCDR1 of SEQ ID NO: 101, an HCDR2 of SEQ ID NO: 115, and an HCDR3 of SEQ ID NO: 128.

[0076] In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 130, an LCDR2 of SEQ ID NO: 153, and an LCDR3 of SEQ ID NO: 175. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 131, an LCDR2 of SEQ ID NO: 154, and an LCDR3 of SEQ ID NO: 176. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 132, an LCDR2 of SEQ ID NO: 155, and an LCDR3 of SEQ ID NO: 177. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 133, an LCDR2 of SEQ ID NO: 156, and an LCDR3 of SEQ ID NO: 178. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 134, an LCDR2 of SEQ ID NO: 157, and an LCDR3 of SEQ ID NO: 179. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 135, an LCDR2 of SEQ ID NO: 158, and an LCDR3 of SEQ ID NO: 180. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 136, an LCDR2 of SEQ ID NO: 159, and an LCDR3 of SEQ ID NO: 181. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 137, an LCDR2 of SEQ ID NO: 160, and an LCDR3 of SEQ ID NO: 182. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 138, an LCDR2 of SEQ ID NO: 161, and an LCDR3 of SEQ ID NO: 183. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 139, an LCDR2 of SEQ ID NO: 162, and an LCDR3 of SEQ ID NO: 184. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 140, an LCDR2 of SEQ ID NO: 163, and an LCDR3 of SEQ ID NO: 185. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 141, an LCDR2 of SEQ ID NO: 164, and an LCDR3 of SEQ ID NO: 186. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 142, an LCDR2 of SEQ ID NO: 165, and an LCDR3 of SEQ ID NO: 187. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 143, an LCDR2 of SEQ ID NO: 166, and an LCDR3 of SEQ ID NO: 188.In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 144, an LCDR2 of SEQ ID NO: 167, and an LCDR3 of SEQ ID NO: 189. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 145, an LCDR2 of SEQ ID NO: 168, and an LCDR3 of SEQ ID NO: 190. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 146, an LCDR2 of SEQ ID NO: 169, and an LCDR3 of SEQ ID NO: 191. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 147, an LCDR2 of SEQ ID NO: 170, and an LCDR3 of SEQ ID NO: 192. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 148, an LCDR2 of SEQ ID NO: 171, and an LCDR3 of SEQ ID NO: 193. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 149, an LCDR2 of SEQ ID NO: 172, and an LCDR3 of SEQ ID NO: 194. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 150, an LCDR2 of SEQ ID NO: 173, and an LCDR3 of SEQ ID NO: 195. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 151, an LCDR2 of SEQ ID NO: 174, and an LCDR3 of SEQ ID NO: 196. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 198, an LCDR2 of SEQ ID NO: 201, and an LCDR3 of SEQ ID NO: 204. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 199, an LCDR2 of SEQ ID NO: 202, and an LCDR3 of SEQ ID NO: 205. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 207, an LCDR2 of SEQ ID NO: 217, and an LCDR3 of SEQ ID NO: 227. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 208, an LCDR2 of SEQ ID NO: 218, and an LCDR3 of SEQ ID NO: 228. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 209, an LCDR2 of SEQ ID NO: 219, and an LCDR3 of SEQ ID NO: 229. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 210, an LCDR2 of SEQ ID NO: 220, and an LCDR3 of SEQ ID NO: 230.In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO:211, an LCDR2 of SEQ ID NO:221, and an LCDR3 of SEQ ID NO:231. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO:212, an LCDR2 of SEQ ID NO:222, and an LCDR3 of SEQ ID NO:232. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO:213, an LCDR2 of SEQ ID NO:223, and an LCDR3 of SEQ ID NO:233. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO:214, an LCDR2 of SEQ ID NO:224, and an LCDR3 of SEQ ID NO:234. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO:215, an LCDR2 of SEQ ID NO:225, and an LCDR3 of SEQ ID NO:235. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO:237, an LCDR2 of SEQ ID NO:245, and an LCDR3 of SEQ ID NO:253. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 238, an LCDR2 of SEQ ID NO: 246, and an LCDR3 of SEQ ID NO: 254. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 239, an LCDR2 of SEQ ID NO: 247, and an LCDR3 of SEQ ID NO: 255. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 240, an LCDR2 of SEQ ID NO: 248, and an LCDR3 of SEQ ID NO: 256. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 241, an LCDR2 of SEQ ID NO: 249, and an LCDR3 of SEQ ID NO: 257. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 242, an LCDR2 of SEQ ID NO: 250, and an LCDR3 of SEQ ID NO: 258. In some embodiments, the anti-CFHR4 antibody comprises an LCDR1 of SEQ ID NO: 243, an LCDR2 of SEQ ID NO: 251, and an LCDR3 comprises the amino acid sequence of SEQ ID NO: 259.

[0077] In some embodiments, the VH of an anti-CFHR4 antibody of the present disclosure comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to any of (a) SEQ ID NOs: 260-265, (b) SEQ ID NOs: 284-293, (c) SEQ ID NOs: 324-334, or (d) SEQ ID NOs: 368-380. In some embodiments, the VL of an anti-CFHR4 antibody of the present disclosure comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to any of (a) SEQ ID NOs: 272-277, (b) SEQ ID NOs: 304-313, (c) SEQ ID NOs: 346-356, or (d) SEQ ID NOs: 394-406.

[0078] In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 260, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 272. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 261, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 273. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 262, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 274.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 263, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 275. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:264, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:276. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 265, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 277.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:284, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:304. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:285, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:305. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:286, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:306.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:287, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:307. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:288, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:308. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:289, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:309.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:290, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:310. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:291, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:311. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:292, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:312.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:293, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:313. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 324, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 346. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 325, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 347. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 99%, or 100% identical) to SEQ ID NO: 326. 348), and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 348. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 327, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 349. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 328, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 350.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 329, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 351. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 330, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 352. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 331, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 353.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 332, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 354. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 333, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 354. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 334, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 356.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 368, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 394. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 369, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 395. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 370, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 396.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 371, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 397. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 372, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 398. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:373, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:399.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 374, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 400. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 375, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 401. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 376, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 402.In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 377, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 403. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 378, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 404. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 379, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 405. In some embodiments, the VH comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 380, and the VL comprises an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 406.

[0079] The nucleic acid or amino acid sequence "identity" described herein can be determined by comparing a nucleic acid or amino acid sequence of interest to a reference nucleic acid or amino acid sequence. Many mathematical algorithms for obtaining optimal alignment and calculating identity between two or more sequences are known and are incorporated into many available software programs. Examples of such programs include CLUSTAL-W, T-Coffee, and ALIGN (for aligning nucleic acid and amino acid sequences), BLAST programs (e.g., BLAST2.1, BL2SEQ, and their later versions), and FASTA programs (e.g., FASTA3x, FAS™, and SSEARCH) (for sequence alignment and sequence similarity search). Sequence alignment algorithms are also described in, for example, Altschul et al., J. Molecular Biol., 215(3):403-410 (1990); Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10):3770-3775 (2009); Durbin et al., eds., Biological Sequence Analysis: Probabilistic Models of Proteins and Nucleic Acids, Cambridge University Press, Cambridge, UK (2009); Soding, Bioinformatics, 21(7):951-960 (2005); Altschul et al., Nucleic Acids Res., 25(17):3389-3402 (1997); and Gusfield, Algorithms on Strings, Trees and Sequences, Cambridge University Press, Cambridge This is disclosed in the UK (1997).

[0080] As will be recognized by those skilled in the art based on the present disclosure, one or more amino acids of the aforementioned anti-CFHR4 antibodies, or antigenic fragments thereof, may be replaced or substituted with different amino acids. An amino acid "replacement" or "substitution" refers to the replacement of one amino acid at a given position or residue with another amino acid at the same position or residue in a polypeptide sequence. Amino acids are broadly classified as "aromatic" or "aliphatic". Aromatic amino acids contain an aromatic ring. Examples of "aromatic" amino acids include histidine (H or His), phenylalanine (F or Phe), tyrosine (Y or Tyr), and tryptophan (W or Trp). Non-aromatic amino acids are broadly classified as "aliphatic". Examples of "aliphatic" amino acids include glycine (G or Gly), alanine (A or Ala), valine (V or Val), leucine (L or Leu), isoleucine (I or Ile), methionine (M or Met), serine (S or Ser), threonine (T or Thr), cysteine ​​(C or Cys), proline (P or Pro), glutamic acid (E or Glu), aspartic acid (A or Asp), asparagine (N or Asn), glutamine (Q or Gln), lysine (K or Lys), and arginine (R or Arg). Aliphatic amino acids can be subdivided into four subgroups. The "large aliphatic non-polar subgroup" consists of valine, leucine, and isoleucine. The "aliphatic less polar subgroup" consists of methionine, serine, threonine, and cysteine. The "aliphatic polar / charged subgroup" consists of glutamic acid, aspartic acid, asparagine, glutamine, lysine, and arginine. The "small residue subgroup" consists of glycine and alanine. The group of charged / polar amino acids can be subdivided into three subgroups: the "positively charged subgroup" consisting of lysine and arginine, the "negatively charged subgroup" consisting of glutamic acid and aspartic acid, and the "polar subgroup" consisting of asparagine and glutamine. The aromatic amino acids can be subdivided into two subgroups: the "nitrogen ring subgroup" consisting of histidine and tryptophan, and the "phenyl subgroup" consisting of phenylalanine and tyrosine.

[0081] Amino acid replacements or substitutions can be conservative, semi-conservative, or non-conservative. The phrase "conservative amino acid substitution" or "conservative mutation" refers to the replacement of one amino acid with another amino acid that has common properties. A functional method for defining common properties between individual amino acids is to analyze the normalized frequency of amino acid changes between corresponding proteins of homologous organisms (Schulz and Schirmer, Principles of Protein Structure, Springer-Verlag, New York (1979)). According to such analysis, groups of amino acids can be defined as being most similar to each other in their effect on the overall protein structure when the amino acids in the group preferentially exchange with each other. Examples of conservative amino acid substitutions include the substitution of amino acids within the above-mentioned subgroups, such as the substitution of arginine with lysine and vice versa, so that a positive charge can be maintained, the substitution of aspartic acid with glutamic acid and vice versa, so that a negative charge can be maintained, the substitution of threonine with serine, so that a free OH can be maintained, and the substitution of asparagine with glutamine, so that a free NH2 can be maintained. "Semi-conservative mutations" include amino acid substitutions of amino acids within the same group listed above, but not within the same subgroup. For example, the substitution of asparagine with aspartic acid, or lysine with asparagine, involves amino acids within the same group but different subgroups. "Non-conservative mutations" involve amino acid substitutions between different groups (e.g., tryptophan to lysine, or serine to phenylalanine, etc.).

[0082] Further, one or more amino acids may be inserted into the anti-CFHR4 antibody, or antigen-binding fragment thereof (e.g., insertion into the heavy and / or light chain variable region amino acid sequence). Any number of suitable amino acids may be inserted into the amino acid sequence of the antibody, or antigen-binding fragment thereof. In this regard, at least one amino acid (e.g., 2 or more, 5 or more, or 10 or more amino acids), but not more than 20 amino acids (e.g., 18 or less, 15 or less, or 12 or less) may be inserted into the amino acid sequence of the antibody, or antigen-binding fragment thereof. For example, 1 to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids) may be inserted into the amino acid sequence of the monoclonal antibody, or antigen-binding fragment thereof. In this regard, the amino acid(s) may be inserted into the antibody, or antigen-binding fragment thereof, at any suitable position. Preferably, the amino acid(s) is inserted into the CDR (e.g., CDR1, CDR2, or CDR3) of the antibody, or antigen-binding fragment thereof.

[0083] The amino acid sequence of anti-CFHR4 antibody or its antigen-binding fragment is not limited to the specific amino acid sequence described herein.In fact, anti-CFHR4 antibody or its antigen-binding fragment can include any heavy or light chain polypeptide that competes with anti-CFHR4 antibody or its antigen-binding fragment for conformational binding to CFHR4.Antibody competition can be assayed using conventional peptide competition assays such as, for example, ELISA, Western blot, or immunohistochemistry (see, for example, U.S. Patents 4,828,981 and 8,568,992, and Braitbard et al., Proteome Sci., 4:12 (2006)).

[0084] The anti-CFHR4 antibodies of the present disclosure may be whole antibodies or antigen-binding fragments of whole antibodies. As defined herein, antigen-binding antibody fragments encompassed by the present disclosure include, but are not limited to, F(ab')2, Fab', Fab, Fv, scFv, dsFv, dAb, and single-chain binding polypeptides. Antibody fragments and their therapeutic utility are further described, for example, in Nelson, AL, MAbs. 2010 Jan-Feb; 2(1): 77-83; Joosten et al., Microbial Cell Factories volume 2, Article number: 1 (2003); and Bates A, Power CA., Antibodies (Basel). 2019; 8(2): 28; doi: 10.3390 / antib8020028). In some embodiments, the anti-CFHR4 antigen-binding fragment is a single-chain variable fragment (scFv), which is an engineered antibody produced by fusion of an immunoglobulin heavy chain (VH) and light chain (VL) via a short polypeptide linker. Single chain variable domain (Fv) fragments (scFv) have been used in the art for a variety of clinical and therapeutic applications, primarily due to their improved pharmacokinetic properties compared to parent monoclonal antibodies and their relative ease of production in large quantities at low cost (Monnier et al., Antibodies 2013, 2(2), 193-208; doi.org / 10.3390 / antib2020193; Safdari et al., Mol Med. 2016; 22:258-270; and Lu, R., Hwang, Y., Liu, I. et al. Development of therapeutic antibodies for the treatment of diseases. J Biomed Sci 27, 1(2020). https: / / doi.org / 10.1186 / s12929-019-0592-z).

[0085] The anti-CFHR4 antibody of the present disclosure may be a diabody. A diabody is an antibody fragment with two antigen binding sites, which may be bivalent or bispecific. See, for example, EP 404,097, WO 1993 / 01161, Hudson et al., Nat.Med.9:129-134(2003), and Hollinger et al., Proc.Natl.Acad.Sci.USA90:6444-6448(1993). Triabodies and tetrabodies are also described in Hudson et al., Nat.Med.9:129-134(2003). The anti-CFHR4 antibody of the present disclosure may be a single domain antibody (also called a nanobody). A single domain antibody is an antibody fragment that includes all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, Mass., see, e.g., U.S. Pat. No. 6,248,516 B1). Antibody fragments can be generated by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies, as well as production by recombinant host cells (e.g., E. coli or phage), as described herein.

[0086] In other embodiments, the anti-CFHR4 antibody is a whole antibody. As defined herein, a whole antibody typically comprises two identical copies of a heavy (H) chain polypeptide and two identical copies of a light (L) chain polypeptide. Each of the heavy chains contains one N-terminal variable (V H ) region and three C-terminal constant (C H1 , C H2 , and C H3 ) region, and each light chain contains one N-terminal variable (V L ) region and one C-terminal constant (C L). The heavy chain C-terminal constant region includes a fragment of the crystallizable (Fc) domain, which determines the class of the antibody and is responsible for humoral and cellular effector functions. Antibodies are classified into five major classes (i.e., "isotypes"), IgG, IgM, IgA, IgD, and IgE, that have different functions in the immune system. IgG is the most abundant immunoglobulin in blood, accounting for 60% of all serum antibodies in humans. IgG antibodies can be subclassified into IgG1, IgG2, IgG3, and IgG4, named in order of abundance in serum (IgG1 being the most abundant) (Vidarsson et al., Frontiers in Immunology. 5:520 (2014)). All anti-CFHR4 monoclonal antibodies described herein can be of any suitable class and / or subclass. In some embodiments, the monoclonal antibody is of the IgG class (e.g., IgG1, IgG2, IgG3, or IgG4). For example, the monoclonal antibody can be an IgG1 antibody.

[0087] As described above, the Fc domain mediates some effector functions of the antibody, such as binding to receptors on target cells and fixation of complement (resulting in effector functions that eliminate antigens). In some embodiments, the Fc domain may be modified or engineered to alter its effector functions. For example, the Fc domain may be modified to improve antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP), as well as to control serum half-life. In some embodiments, the Fc domain of an anti-CFHR4 antibody may be engineered to modulate affinity for Fc receptors, such as Fcγ receptors (FcγR) and fetal Fc receptors (FcRn). Indeed, optimization of the interaction between antibodies and FcγR has emerged as a promising approach to enhance the activity of therapeutic antibodies for the treatment of various diseases (Mimoto et al., Curr. Pharm. Biotechnol. 17, 1298-1314 (2016); Lazar et al., Proc. Natl Acad. Sci. USA 103, 4005-4010 (2006); Richards et al., Mol. Cancer Ther. 7, 2517-2527 (2008); Nordstrom et al., Breast Cancer Res. 13, R123 (2011); and Kang, TH, Jung, ST, Exp Mol Med 51, 1-9 (2019)). The Fc domain can also be modified to improve serum half-life, for example, by engineering IgG Fc for higher FcRn binding (Zalevsky et al., Nat. Biotechnol. 28, 157-159 (2010), and Dall'Acqua et al., J. Immunol. 169, 5171-5180 (2002)). In other embodiments, the Fc domain can be engineered to create monovalency or bispecificity of the antibody to improve therapeutic efficacy. For example, an Fc domain can be generated that does not form homodimers but remains as a soluble monomeric mFc that exhibits high affinity for FcγRI but has no detectable binding to FcγRIIIa.In other embodiments, heterodimeric Fc domains can be generated to obtain bispecific properties of antigen binding that avoid homodimer formation. Engineered Fc domains can be generated by introducing point mutations or by altering the glycosylation of the Fc domain (Saunders, KO, Front Immunol. 2019; 10: 1296; Kelley, RF, Meng, YG, Liu et al., J Biol Chem. 2014; 289: 3571-90; Monnet et al., MAbs. 2014; 6: 422-36; Li et al., Proc Natl Acad Sci USA. 2017; 114: 3485-90; and Lin et al., Proc Natl Acad Sci USA. 2015; 112: 10611-6; Kang and Jung, supra).

[0088] Multispecific anti-CFHR4 antibody As described above, the anti-CFHR4 antibodies of the present disclosure can be monoclonal, human, humanized, and / or chimeric antibodies. In some embodiments, the antibody is a fragment selected from the group consisting of Fab, Fab-C, Fab'-SH, Fv, scFV, and (Fab')2 fragments. In some embodiments, the anti-CFHR4 antibody is a monospecific antibody. In some embodiments, the anti-CFHR4 antibody is a bispecific antibody. In some embodiments, the anti-CFHR4 antibody comprises two or more single domain antibodies forming a bivalent, trivalent, or tetravalent antibody that recognizes different epitopes on the same or different antigens.

[0089] In some embodiments, the anti-CFHR4 antibody provided herein is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567, and Morrison et al., Proc. Natl. Acad. Sci. USA. 81:6851-6855 (1984). In one example, the chimeric antibody comprises a non-human variable region (e.g., a variable domain from a mouse, rat, hamster, rabbit, or a non-human primate, e.g., a monkey) and a human constant domain. In a further example, the chimeric antibody is a "class-switched" antibody, in which the class or subclass is changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0090] In certain embodiments, a chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVRs, e.g., CDRs (or portions thereof) are derived from a non-human antibody and the FRs (or portions thereof) are derived from a human antibody sequence. Optionally, the humanized antibody also comprises at least a portion of a human constant region. In some embodiments, some FR residues in the humanized antibody are replaced with the corresponding residues from the non-human antibody (e.g., the antibody from which the HVR residues are derived), e.g., to restore or improve antibody specificity or affinity.

[0091] Humanized antibodies and methods for making them are reviewed, e.g., in Almagro and Fransson, Front. Biosci. 13:1619-1633, (2008), and are also described in, e.g., 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) (describing specificity determining region (SDR) grafting), Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"), Dall'Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"), Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing a "guided selection" approach to FR shuffling).

[0092] According to the above embodiment, the anti-CFHR4 antibody of the present disclosure can be made into a bivalent, trivalent, or tetravalent format. For example, the anti-CFHR4 antibody of the present disclosure can be a bivalent bispecific antibody with a heteromeric heavy chain (e.g., triomab, knob-into-hole (KIH), duobody, etc.). The anti-CFHR4 antibody of the present disclosure can be a tetravalent multispecific antibody composed of an IgG with other binding domains fused to either the N-terminus or C-terminus of either the heavy or light chain (e.g., dual variable domain [DVD], IgG-scFv fusion, mabtyrin (an IgG with a non-antibody binding scaffold "sentirin" fused to the C-terminus of the heavy chain). The anti-CFHR4 antibody of the present disclosure can be composed of an IgG with additional antigen binding sites added into the structure (e.g., two-in-one antibody, MAT "Modular Antibody Technology" platform from F-Star). The anti-CFHR4 antibodies of the present disclosure can be engineered antibody fragments linked by short peptide linkers that can be made into bivalent, trivalent, or tetravalent formats addressing two to three targets (e.g., bispecific T cell engagers (BiTEs), nanobody platforms, dual affinity retargeting (DART) antibodies, "tandem antibody" structures (TandAbs)). The anti-CFHR4 antibodies of the present disclosure can also be composed of chemically conjugated IgG.

[0093] In some embodiments, the anti-CFHR4 antibodies of the present disclosure are multispecific antibodies, such as bispecific antibodies, that have binding specificities for at least two different antigens. In some embodiments, the anti-CFHR4 antibodies of the present disclosure, or antigen-binding fragments thereof, can be used to form one arm (e.g., antigen-binding portion) of a bispecific antibody, while the other arm of the bispecific antibody can be specific for a different antigen. In some embodiments, the other antigens include interleukin-1 beta (IL-1β), interleukin-6 (IL-6); interleukin-6 receptor (IL-6R); interleukin-13 (IL-13); IL-13 receptor (IL-13R); PDGF (e.g., PDGF-BB); angiopoietin; angiopoietin 2 (Ang2); Tie2; S1P; integrins αvβ3, αvβ5, and α5β1; betacellulin; apelin / APJ; erythropoietin; complement factor D; TNFα; HtrA1; VEGF receptors (e.g., VEGFR1, VEGFR2, VEGFR3, VEGFR4, VEGFR5, VEGFR6, VEGFR7, VEGFR8, VEGFR9, VEGFR11, VEGFR12, VEGFR13, VEGFR14, VEGFR15, VEGFR16, VEGFR17, VEGFR18, VEGFR19, VEGFR20, VEGFR210, VEGFR22, VEGFR23, VEGFR24, VEGFR25, VEGFR26, VEGFR27, VEGFR28, VEGFR29, VEGFR30, VEGFR31, VEGFR32, VEGFR33, VEGFR34, VEGFR35, VEGFR36, VEGFR37, VEGFR38, VEGFR39, VEGFR39, V VEGF receptor 1, VEGFR2, VEGFR3, membrane-bound VEGF receptor (mbVEGFR) or soluble VEGF receptor (sVEGFR)); ST-2 receptor; and proteins genetically related to age-related macular degeneration (AMD) risk, such as complement pathway components C2, factor B, factor H, CFHR3, C3b, C5, C5a, and C3a; HtrA1; ARMS2; TIMP3; HLA; interleukin-8 (IL-8); CX3CR1; TLR3; TLR4; CETP; LIPC; COL10A1; and TNFRSF10A.In other embodiments, the bispecific antibody is selected from the group consisting of CFHR4 and IL-1β, CFHR4 and IL-6, CFHR4 and IL-6R, CFHR4 and IL-13, CFHR4 and IL-13R, CFHR4 and PDGF (e.g., PDGF-BB), CFHR4 and angiopoietin, CFHR4 and Ang2, CFHR4 and Tie2, CFHR4 and S1P, CFHR4 and integrin αvβ3, CFHR4 and integrin αvβ5, CFHR4 and integrin α5β1, CFHR4 and betacellulin, CFHR4 and apelin / APJ, CFHR4 and erythropoietin, CFHR4 and complement factor D, CFHR4 and TNFα, CFHR4 and HtrA1, CFHR4 and VEGF receptors. The antibody may have binding specificity for a receptor (e.g., VEGFR1, VEGFR2, VEGFR3, mbVEGFR, or sVEGFR), CFHR4 and the ST-2 receptor, CFHR4 and C2, CFHR4 and factor B, CFHR4 and factor H, CFHR4 and CFHR3, CFHR4 and C3b, CFHR4 and C5, CFHR4 and C5a, CFHR4 and C3a, CFHR4 and ARMS2, CFHR4 and TIMP3, CFHR4 and HLA, CFHR4 and IL-8, CFHR4 and CX3CR1, CFHR4 and TLR3, CFHR4 and TLR4, CFHR4 and CETP, CFHR4 and LIPC, CFHR4 and COL10A1, or CFHR4 and TNFRSF10A.

[0094] In some embodiments, a bispecific antibody of the present disclosure comprises an anti-CFHR4 antibody, or an antigen-binding fragment thereof, and an anti-VEGF antibody, or an antigen-binding fragment thereof. Such bispecific antibodies can be used to target the complement activation pathway from two different mechanisms, thus providing additional therapeutic benefits. For example, the anti-CFHR4 arm can be any of the anti-CFHR4 antibodies of the present disclosure, and the anti-VEGF arm can be any VEGF antagonist, including, but not limited to, anti-VEGF antibodies (e.g., bevacizumab, sevacizumab, and ranibizumab), anti-VEGFR2 antibodies and related molecules (e.g., ramucirumab, tanibirumab, aflibercept), anti-VEGFR1 antibodies and related molecules (e.g., icrucumab, aflibercept (VEGF Trap-Eye, EYLEA®), and ziv-aflibercept (VEGF Trap, ZALTRAP®)), the anti-VEGF arm (including the anti-VEGF, anti-VEGFR1, and anti-VEGFR2 arms) of a VEGF bispecific antibody (e.g., MP-0250, vanucizumab (VEGF-ANG2)).

[0095] In other embodiments, the bispecific antibody of the present disclosure comprises an anti-CFHR4 antibody, or an antigen-binding fragment thereof, and an anti-C3 antibody, or an antigen-binding fragment thereof. Such bispecific antibodies can be used to target complement activation pathways from two different mechanisms, thus providing additional therapeutic effects. For example, the anti-CFHR4 arm can be any of the anti-CFHR4 antibodies of the present disclosure, and the anti-C3 arm can be any C3 antagonist, including but not limited to POT-4 (AL-78898A), APL-2, and NGM621.

[0096] In other embodiments, the bispecific antibody of the present disclosure comprises an anti-CFHR4 antibody, or an antigen-binding fragment thereof, and an anti-C5 antibody, or an antigen-binding fragment thereof. For example, the anti-CFHR4 arm can be any of the anti-CFHR4 antibodies of the present disclosure, and the anti-C5 arm can be any C5 antagonist, including but not limited to eculizumab and tesidormab (LFG316). In other embodiments, the bispecific antibody of the present disclosure comprises an anti-CFHR4 antibody, or an antigen-binding fragment thereof, and an anti-FD antibody, or an antigen-binding fragment thereof. For example, the anti-CFHR4 arm can be any of the anti-CFHR4 antibodies of the present disclosure, and the anti-C5 arm can be any C5 antagonist, including but not limited to lampalizumab.

[0097] Functional characterization of anti-CFHR4 antibodies In accordance with the above embodiments, the present disclosure provides anti-CFHR4 antibodies comprising various functional characteristics. In some embodiments, the anti-CFHR4 antibodies described herein bind to an antigen on CFHR4 (SEQ ID NO: 580), or a variant or isoform thereof, via interaction with an antigenic determinant (epitope) thereof. In some embodiments, binding of the anti-CFHR4 antibody to CFHR4 reduces complement activation. In some embodiments, the anti-CFHR4 antibody binds to human CFHR4b and / or binds to human CFHR4b. D It binds at approximately 100 nM or less (Figure 6).

[0098] In some embodiments, the antibody cross-reacts with cynomolgus monkey CFHR4b protein (cCFHR4b). In some embodiments, the antibody cross-reacts with ATX-P-560, ATX-P-561, ATX-P-562, ATX-P-563, ATX-P-564, ATX-P-565, ATX-P-566, ATX-P-568, ATX-P-569, ATX-P-570, ATX-P-571, ATX-P-573, ATX-P-574, ATX-P-576, ATX-P-577, ATX-P-578, ATX-P- 579, ATX-P-580, ATX-P-581, ATX-P-582, ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-591, ATX-P-592, ATX-P-594, ATX-P-596, ATX-P-600, and ATX-P-604 (Figure 7).

[0099] In some embodiments, the antibody does not cross-react with cynomolgus CFHR4b protein (cCFHR4b). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and the VL region of an antibody selected from the group consisting of ATX-P-572, ATX-P-595, ATX-P-597, ATX-P-598, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-607, ATX-P-608, ATX-P-609, and ATX-P-610 (FIG. 7).

[0100] In some embodiments, the antibody cross-reacts with complement factor H related protein 4a (CFHR4a). In some embodiments, the antibody cross-reacts with complement factor H related protein 4a (CFHR4a). In some embodiments, the antibody cross-reacts with complement factor H related protein 4a (CFHR4a). 583, ATX-P-587, ATX-P-588, ATX-P-591, ATX-P-592, ATX-P-594, ATX-P-595, ATX-P-596, ATX-P-597, ATX-P-600, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-604, ATX-P-607, ATX-P-608, ATX-P-609, and ATX-P-610 (FIG. 8).

[0101] In some embodiments, the antibody does not cross-react with complement factor H related protein 4a (CFHR4a). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to ATX-P-598 (Figure 8).

[0102] In some embodiments, the antibody cross-reacts with complement factor H related protein 3 (CFHR3). In some embodiments, the antibody cross-reacts with complement factor H related protein 3 (CFHR3). In some embodiments, the antibody cross-reacts with complement factor H related protein 3 (CFHR3). and VL regions that are at least 90% identical to the VH and VL regions of an antibody selected from the group consisting of ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-592, ATX-P-594, ATX-P-595, ATX-P-596, ATX-P-600, ATX-P-603, ATX-P-604, ATX-P-608, ATX-P-609, and ATX-P-610 (Figure 9).

[0103] In some embodiments, the antibody does not cross-react with complement factor H related protein 3 (CFHR3). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and the VL region of an antibody selected from the group consisting of ATX-P-560, ATX-P-570, ATX-P-571, ATX-P-582, ATX-P-591, ATX-P-597, ATX-P-598, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-604, and ATX-P-607 (Figure 9).

[0104] In some embodiments, the antibody cross-reacts with complement factor H related protein 1 (CFHL1). In some embodiments, the antibody cross-reacts with complement factor H related protein 1 (CFHL1). In some embodiments, the antibody cross-reacts with complement factor H related protein 1 (CFHL1). 580, ATX-P-581, ATX-P-582, ATX-P-583, ATX-P-587, ATX-P-588, ATX-P-591, ATX-P-594, ATX-P-595, ATX-P-596, ATX-P-600, and ATX-P-604 (Figure 10).

[0105] In some embodiments, the antibody does not cross-react with complement factor H related protein 1 (CFHL1). In some embodiments, the antibody comprises a VH region and a VL region that are at least 90% identical to the VH region and the VL region of an antibody selected from the group consisting of ATX-P-560, ATX-P-569, ATX-P-572, ATX-P-592, ATX-P-597, ATX-P-598, ATX-P-601, ATX-P-602, ATX-P-603, ATX-P-607, ATX-P-608, ATX-P-609, and ATX-P-610 (Figure 10).

[0106] Embodiments of the present disclosure also include means for evaluating one or more functional and / or biochemical characteristics of the anti-CFHR4 antibodies described herein. In one embodiment, the present disclosure provides a C3 convertase assembly assay (FIGS. 11A-11C). The C3 convertase assembly assay was developed as an antibody screening assay to evaluate the ability of the CFHR4 antibodies of the present disclosure to affect C3 convertase formation (FIG. 11A). More specifically, C3bBb is the C3 convertase (EC 3.4.21.43) of the alternative pathway of complement activation. The C3 enzyme is involved in the amplification of pathway activation and deposition of C3b and the membrane attack complex on target cells. It is regulated by serum proteins factor H, factor I (EC 3.4.21.45), and properdin. Formation of C3 convertase requires C3b, factor B, factor D (EC 3.4.21.46), and Mg2+. After formation of the reversible bimolecular complex C3bB(Mg), factor D cleaves factor B, releasing the activation fragment Ba and generating the active enzyme C3bBb. The C3 enzyme is a serine protease whose catalytic site resides in the Bb subunit. In addition, CFHR4 has been reported to directly promote C3 convertase formation by binding to C3b, and it has a higher ability to assemble C3bBb on CFHR4, and the C3bBb-CFHR4 complex is more resistant to CFH cleavage compared to C3bBb.

[0107] Therefore, the C3 convertase assay of the present disclosure is designed to test the ability of CHFR4 antibodies to disrupt the formation of the C3 convertase complex by measuring the amount of factor B present using anti-FB monoclonal antibodies conjugated to a reporter moiety.As illustrated in Figure 11A, in the presence of CFHR4 (in the absence of an effective CFHR4 antibody), the C3 convertase complex is formed, the anti-FB antibody binds to FB, and a signal is detected (i.e., complement activation).However, in the presence of an effective anti-CFHR4 antibody, the C3 convertase complex is disrupted, the FB bound to the antibody is washed away, and the signal is reduced (i.e., no complement activation).

[0108] As shown in FIG. 11B, dose-response curves were generated for the top 8 inhibitory antibodies screened in the C3 convertase assembly assay. Percent C3 convertase activity is plotted relative to the negative control (diluent only). FIG. 11C includes IC50 values ​​calculated for each antibody tested in FIG. 11B. Thus, according to these assay results, certain anti-CFHR4 antibodies of the present disclosure attenuate C3 convertase activity. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO: 330, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 352. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 313. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO: 378, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO: 404. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO:377, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:403. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO:376, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:402. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO:289, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:309. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO:371, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:397. In some embodiments, the VH of the anti-CFHR4 antibody comprises an amino acid sequence at least 90% identical to SEQ ID NO:284, and the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO:304 (FIGS. 11A-11C).

[0109] Polypeptides and Expression Vectors Embodiments of the present disclosure also include polynucleotides encoding any of the anti-CFHR4 antibodies of the present disclosure. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 70% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0110] In some embodiments, the polynucleotide comprises a sequence that is at least 75% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 75% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 75% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 75% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0111] In some embodiments, the polynucleotide comprises a sequence that is at least 80% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 80% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 80% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 80% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0112] In some embodiments, the polynucleotide comprises a sequence that is at least 85% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 85% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 85% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 85% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0113] In some embodiments, the polynucleotide comprises a sequence that is at least 90% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 90% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 90% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 90% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0114] In some embodiments, the polynucleotide comprises a sequence that is at least 95% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 381-393. In some embodiments, the polynucleotide comprises a sequence that is at least 95% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419. In some embodiments, the polynucleotide comprises a sequence that is at least 95% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 266-271, (b) SEQ ID NOs: 294-303, (c) SEQ ID NOs: 335-345, or (d) SEQ ID NOs: 380-393. In some embodiments, the polynucleotide comprises a sequence that is at least 95% identical to any of the following nucleic acid sequences: (a) SEQ ID NOs: 278-283, (b) SEQ ID NOs: 314-323, (c) SEQ ID NOs: 357-367, or (d) SEQ ID NOs: 407-419.

[0115] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises (a) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:266 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:278; (b) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:278; 67 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:279; (c) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:268; 280; (d) nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:269; 281, (e) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:270 and at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:282.or (f) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 270 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 283.

[0116] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises (a) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:294 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:314; (b) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:294; 95 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 315; (c) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 296; (d) nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 316, (e) nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 297, 317, (e) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:298 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:318.(f) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 299 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 319; (g) (h) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 300 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 320; (i) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 301; 321, (i) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 321, (ii) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 302, and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 322, or (j) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 303 and at least 70% identical (e.g., at least 70%, at least 75%,at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to the nucleic acid sequence of

[0117] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises (a) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 335 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 357; (b) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 357; 36 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 358; (c) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 337; 359; (d) nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 338; 360, (e) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 339 and at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 361.(f) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 340 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 362; (g) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 362; (h) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:341 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:363; 364, (i) nucleic acid sequences and sequence numbers that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 364, (ii) nucleic acid sequences and sequence numbers that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 343, (j) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 344 and at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 366.or (k) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical) to SEQ ID NO: 345 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical) to SEQ ID NO: 367.

[0118] In some embodiments, a polynucleotide encoding an anti-CFHR4 antibody of the disclosure comprises (a) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:381 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:407; (b) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:3 82 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 408; (c) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 383; (d) nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 384 and nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 384. 410, (e) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 385 and at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 411.(f) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 386 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 412; (g) a nucleic acid sequence (h) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:387 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:413; (i) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:388; , at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 414 and nucleic acid sequences that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 414, (i) nucleic acid sequences and sequence numbers that are at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 389. (j) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:390 and at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO:416.(k) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical) to SEQ ID NO: 391 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical) to SEQ ID NO: 417; (l) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical) to SEQ ID NO: 392; 5%, or 100% identical) to SEQ ID NO: 418 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 418; or (m) a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 393 and a nucleic acid sequence that is at least 70% identical (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical) to SEQ ID NO: 419.

[0119] In accordance with these embodiments, the present disclosure includes an expression vector comprising any of the polynucleotides encoding the anti-CFHR4 antibodies of the present disclosure. In some embodiments, the expression vector is suitable for producing the anti-CFHR4 antibodies of the present disclosure for delivering the antibodies to a subject. In certain embodiments, the nucleic acid sequence is in the form of a vector. The vector can be, for example, a plasmid, an episome, a cosmid, a viral vector (e.g., a retrovirus or an adenovirus), or a phage. Suitable vectors and vector preparation methods are well known in the art (see, for example, Sambrook et al., Molecular Cloning, a Laboratory Manual, 4th edition, Cold Spring Harbor Press, Cold Spring Harbor, NY (2012), and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, New York, NY (1994)).

[0120] In addition to the nucleic acid encoding the anti-CFHR4 antibody or antigen-binding fragment thereof, the vector desirably contains expression control sequences, such as a promoter, enhancer, polyadenylation signal, transcription terminator, internal ribosome entry site (IRES), etc., that provide for expression of the nucleic acid sequence encoding the antibody in a host cell. Exemplary expression control sequences are known in the art and described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology, Vol. 185, Academic Press, San Diego, Calif. (1990).

[0121] A vector containing a nucleic acid sequence encoding an anti-CFHR4 antibody or antigen-binding fragment thereof can be introduced into a host cell (including any suitable prokaryotic or eukaryotic cell) capable of expressing the polypeptide encoded thereby. Examples of suitable prokaryotic cells include, but are not limited to, cells from the genera Bacillus (such as Bacillus subtilis and Bacillus brevis), Escherichia (such as E. coli), Pseudomonas, Streptomyces, Salmonella, and Erwinia. Particularly useful prokaryotic cells include various strains of Escherichia coli (e.g., K12, HB101 (ATCC No. 33694), DH5α, DH10, MC1061 (ATCC No. 53338), and CC102). Suitable eukaryotic cells are known in the art and include, for example, yeast cells, insect cells, and mammalian cells. Examples of suitable yeast cells include those from the genera Hansenula, Kluyveromyces, Pichia, Rhinosporidium, Saccharomyces, and Schizosaccharomyces. Suitable insect cells include Sf-9 and HIS cells (Invitrogen, Carlsbad, Calif.), as described, for example, in Kitts et al., Biotechniques, 14:810-817 (1993), Lucklow, Curr. Opin. Biotechnol., 4:564-572 (1993), and Lucklow et al., J. Virol., 67:4566-4579 (1993). Examples of suitable mammalian cells include, but are not limited to, Chinese hamster ovary cells (CHO) (ATCC No. CCL61), CHO DHFR cells (Urlaub et al., Proc. Natl. Acad. Sci. USA, 97:4216-4220 (1980)), human embryonic kidney (HEK) 293 or 293T cells (ATCC No. CRL1573), and 3T3 cells (ATCC No. CCL92).Other suitable mammalian cell lines are the monkey COS-1 cell line (ATCC No. CRL1650) and COS-7 cell line (ATCC No. CRL1651), and the CV-1 cell line (ATCC No. CCL70). Further exemplary mammalian host cells include primate and rodent cell lines, including transformed cell lines. Also suitable are normal diploid cells, cell lines derived from in vitro culture of primary tissues, and primary explants. Other suitable mammalian cell lines include, but are not limited to, mouse neuroblastoma N2A cells, HeLa, mouse L-929 cells, and BHK or HaK hamster cell lines, all available from ATCC. Methods for selecting suitable mammalian host cells, as well as methods for transforming, culturing, amplifying, screening, and purifying such cells are well known in the art (see, e.g., Ausubel et al., eds., Short Protocols in Molecular Biology, 5th ed., John Wiley & Sons, Inc., Hoboken, NJ (2002)). Preferably, the mammalian cells are human cells.

[0122] In some embodiments, a vector may include a means for attaching a detection moiety to an anti-CFHR4 antibody of the disclosure. In some embodiments, a vector may include a means for attaching a purification moiety to an anti-CFHR4 antibody of the disclosure. Exemplary detection and / or purification moieties / tags that may be attached to an anti-CFHR4 antibody of the disclosure include, but are not limited to, hemagglutinin (HA), c-Myc, V5, DYKDDDDK, His tags (e.g., 6x-HIS), glutathione S-transferase (GST), maltose binding protein (MBP), fluorophores (e.g., green fluorescent protein (GFP), red fluorescent protein (RFP), mCherry, chromophores, and / or luminescent peptides (e.g., luciferase).

[0123] In some embodiments, the expression vector is suitable for use in gene therapy (e.g., an expression vector for delivering a polynucleotide encoding an anti-CFHR4 antibody of the present disclosure to a subject). In some embodiments, the expression vector is a herpes simplex virus (HSV) vector, or a retroviral vector. In some embodiments, the expression vector is an adeno-associated virus (AAV) vector, or includes an AAV backbone. For example, AAV vectors have been designed, produced, and used to mediate gene delivery in human subjects, including for therapeutic purposes. Typically, AAV vectors for use in gene transfer comprise a replication-deficient AAV genome that lacks functional Rep and Cap coding viral sequences. Such replication-deficient AAV vectors more preferably lack most or all of the Rep and Cap coding sequences, and essentially retain one or two AAV ITR sequences and packaging sequences. The defective genome is packaged into a viral particle to form a defective recombinant AAV virus, also referred to as an "AAV vector." Methods for producing such AAV vectors have been disclosed in the literature, including the use of packaging cells, accessory viruses or plasmids, and / or baculovirus systems (Samulski et al., (1989) J. Virology 63, 3822; Xiao et al., (1998) J. Virology 72, 2224; Inoue et al., (1998) J. Virol. 72, 7024; WO98 / 22607; WO2005 / 072364). Methods for generating pseudotyped AAV vectors (e.g., WO00 / 28004) have also been reported, as well as various modifications or formulations of AAV vectors to reduce immunogenicity during in vivo administration (e.g., see WO01 / 23001, WOOO / 73316, WO04 / 112727, W005 / 005610, WO99 / 06562). AAV vectors can be prepared or derived from various serotypes of AAV, which can further be mixed with or with other types of viruses to generate chimeric (e.g., pseudotyped) AAV viruses.Examples of tAAV are human AAV4 vectors, human AAV7 vectors, human AAV9 vectors, human AAV10 vectors, or bovine AAV vectors. AAV vectors can be derived from a single AAV serotype, or can contain sequences or components from at least two different AAV serotypes (pseudotyped AAV vectors), such as an AAV vector that contains an AAV genome from one AAV serotype (e.g., AAV9) and a capsid that is at least partially derived from a separate AAV serotype. An AAV vector as used herein is a vector that contains at least one component that can be derived from an adeno-associated virus. Preferably, the component is involved in the biological mechanism by which the vector infects or transduces target cells and expresses the anti-CFHR4 antibody of the present disclosure (e.g., ocular delivery / expression).

[0124] In other embodiments, the expression vector is a lentiviral vector (LV) or comprises a LV backbone. Lentiviruses are part of the larger retrovirus group. A detailed list of lentiviruses can be found in Coffin et al (1997) "Retroviruses" Cold Spring Harbour Laboratory Press Eds: JM Coffin, SM Hughes, HE Varmus pp 758-763). For example, lentiviruses can be divided into primate and non-primate groups. Examples of primate lentiviruses include, but are not limited to, human immunodeficiency virus (HIV), the causative agent of human autoimmune deficiency syndrome (AIDS), and simian immunodeficiency virus (SIV). The non-primate lentivirus group includes the prototype "slow-release virus" Visna / Maeji virus (VMV), as well as the related Caprine Arthritis Encephalitis Virus (CAEV), Equine Infectious Anemia Virus (EIAV), Feline Immunodeficiency Virus (FIV), Maedi Visna Virus (MVV) and Bovine Immunodeficiency Virus (BIV). In one embodiment, the lentivirus vector is derived from HIV-1, HIV-2, SIV, FIV, BIV, EIAV, CAEV, or Visna lentivirus. The lentivirus family differs from retroviruses in that lentiviruses have the ability to infect both dividing and non-dividing cells (Lewis et al (1992) EM BO J 11(8):3053-3058 and Lewis and Emerman (1994) J Virol 68(1):510-516). In contrast, other retroviruses, such as MLV, are unable to infect non-dividing or slowly dividing cells, such as cells that make up, for example, muscle, brain, lung, and liver tissue. As used herein, a lentiviral vector is a vector that includes at least one component part that can be derived from a lentivirus. Preferably, that component part is involved in the biological mechanism by which the vector infects or transduces target cells and expresses the anti-CFHR4 antibodies of the present disclosure (e.g., ocular delivery / expression).

[0125] Additional compositions and methods for ocular gene therapy can be found, for example, in Bordet, T., and Behar-Cohen, F., "Ocular gene therapies in clinical practice: viral vectors and nonviral alternatives," Drug Discovery Today, Volume 24, Issue 8, August 2019, Pages 1685-1693. In some embodiments, gene therapy platforms, methods, and compositions that can be used to deliver (e.g., ocular delivery) the anti-CFHR4 antibodies of the present disclosure to a subject include those platforms, methods, and compositions disclosed in US20220025396, US20220011308, US20210371877, US20210363192, US20190078099, US20190038724, and US10494646B2, which are incorporated herein by reference. In other embodiments, gene therapy platforms, methods, and compositions that can be used to deliver the anti-CFHR4 antibodies of the present disclosure to a subject (e.g., ocular delivery) include platforms, methods, and compositions based on HMR59 (Hemera Biosciences), which blocks the membrane attack complex formed during the final step of the complement cascade via its protein product, soluble CD59. HMR59 is designed to be administered as a single intraocular injection.

[0126] In accordance with these embodiments, the disclosure also provides methods of administering ocular gene therapy to a subject in need thereof, comprising injecting a pharmaceutical composition comprising an effective amount of an expression vector described herein (e.g., an expression vector comprising a polynucleotide encoding an anti-CFHR4 antibody of the disclosure). As described further below, the disclosure also provides methods of treating AMD and / or GA, comprising administering a pharmaceutical composition comprising an effective amount of an expression vector described herein (e.g., an expression vector comprising a polynucleotide encoding an anti-CFHR4 antibody of the disclosure). In some embodiments, administering the pharmaceutical composition treats at least one symptom of AMD and / or GA.

[0127] Pharmaceutical compositions and methods of treatment The anti-CFHR4 antibodies of the present disclosure can be administered as part of a pharmaceutical composition in a therapeutically effective amount to treat ocular diseases (e.g., AMD or GA). In some embodiments, the composition is suitable for ocular administration. In some embodiments, ocular administration includes injection into the vitreous humor. In some embodiments, ocular administration includes delivering the antibody using a conjunctival insert, contact lens, gel, nanoparticles, mucoadhesive polymers, ointments, solutions, suspensions, eye drops, and / or implants (e.g., Susvimo™). Recent methods and formulations for ocular administration can be found, for example, in Souto, EB, et al. “Advanced Formulation Approaches for Ocular Drug Delivery: State-Of-The-Art and Recent Patents,” Pharmaceutics, 2019 Sep;11(9):460.

[0128] According to these embodiments, the method comprises administering a pharmaceutical composition comprising a therapeutically effective amount of an anti-CFHR4 antibody of the present disclosure. In some embodiments, the pharmaceutical composition is administered to the eye to treat at least one AMD symptom. In some embodiments, the AMD comprises wet AMD. In some embodiments, the AMD comprises dry AMD (e.g., GA). In some embodiments, the at least one AMD symptom comprises visual distortion, reduced central vision, blurred vision, and / or difficulty adapting to low light. In some embodiments, administering the pharmaceutical composition reduces complement activation in the subject's eye.

[0129] In some embodiments, a pharmaceutical composition comprising a therapeutically effective amount of an anti-CFHR4 antibody of the present disclosure is administered at a dose ranging from about 0.0001 mg / dose to about 100 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.001 mg / dose to about 100 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.01 mg / dose to about 100 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.1 mg / dose to about 100 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 1.0 mg / dose to about 100 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 10 mg / dose to about 100 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / dose to about 10 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / dose to about 1.0 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / dose to about 0.1 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / dose to about 0.001 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.01 mg / dose to about 10 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.001 mg / dose to about 1.0 mg / dose. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.1 mg / dose to about 10 mg / dose.

[0130] In some embodiments, a pharmaceutical composition comprising a therapeutically effective amount of an anti-CFHR4 antibody of the present disclosure is administered at a dose ranging from about 0.0001 mg / ml to about 100 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.001 mg / ml to about 100 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.01 mg / ml to about 100 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.1 mg / ml to about 100 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 1.0 mg / ml to about 100 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 10 mg / ml to about 100 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / ml to about 10 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / ml to about 1.0 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / ml to about 0.1 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / ml to about 0.01 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.0001 mg / ml to about 0.001 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.01 mg / ml to about 10 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.001 mg / ml to about 1.0 mg / ml. In some embodiments, the anti-CFHR4 antibody is administered at a dose ranging from about 0.1 mg / ml to about 10 mg / ml.

[0131] As used herein, the terms "treatment", "treating" and the like refer to obtaining a desired pharmacological and / or physiological effect. In some embodiments, the effect is therapeutic, i.e., the effect partially or completely cures the disease and / or adverse symptoms resulting from the disease. To this end, the methods of the present disclosure include administering a "therapeutically effective amount" of an anti-CFHR4 antibody, or a composition comprising an anti-CFHR4 antibody. A "therapeutically effective amount" refers to an amount effective, at the dosage and for the period of time necessary, to achieve a desired therapeutic result. A therapeutically effective amount may vary depending on factors such as the disease state, age, sex, and weight of the individual, as well as the ability of the monoclonal antibody to elicit a desired response in the individual. For example, a therapeutically effective amount of an anti-CFHR4 antibody of the present disclosure is an amount that treats at least one AMD and / or GA symptom in a subject. In some embodiments, the pharmacological and / or physiological effect may be prophylactic, i.e., the effect completely or partially prevents a disease or its symptoms. In this regard, the methods of the present disclosure include administering a "prophylactically effective amount" of an anti-CFHR4 antibody, or a composition comprising an anti-CFHR4 antibody. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired prophylactic result (eg, prevention of the onset of AMD and / or GA).

[0132] A typical therapeutically effective dose of an anti-CFHR4 antibody of the disclosure can range, for example, from about 0.0001 mg / dose to about 100 mg / dose for each eye being treated. In some embodiments, a therapeutically effective dose of an anti-CFHR4 antibody of the disclosure can range from about 0.001 mg / dose to about 100 mg / dose, about 0.01 mg / dose to about 100 mg / dose, about 0.05 mg / dose to about 50 mg / dose, about 0.1 mg / dose to about 10 mg / dose, about 0.5 mg / dose to about 5 mg / dose, and about 1 mg / dose to about 10 mg / dose. In some embodiments, a therapeutically effective concentration of an anti-CFHR4 antibody of the disclosure can be, for example, from about 0.0001 mg to about 100 mg of antibody per milliliter of solution. In some embodiments, a therapeutically effective concentration of an anti-CFHR4 antibody of the disclosure may range from about 0.001 mg / ml to about 100 mg / ml, about 0.01 mg / ml to about 100 mg / ml, about 0.1 mg / ml to about 100 mg / ml, about 1.0 mg / ml to about 100 mg / ml, about 0.001 mg / ml to about 50 mg / ml, about 0.01 mg / ml to about 50 mg / ml, 0.1 mg / ml to about 50 mg / ml, about 0.1 mg / ml to about 25 mg / ml, about 0.1 mg / ml to about 10 mg / ml, and about 1.0 mg / ml to about 10 mg / ml. In some embodiments, a therapeutically effective dose of an anti-CFHR4 antibody of the disclosure may be exactly or approximately 0.1 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.35 mg, 0.4 mg, 0.45 mg, 0.5 mg, 0.55 mg, 0.6 mg, 0.65 mg, 0.7 mg, 0.75 mg, 0.8 mg, 0.85 mg, 0.9 mg, 0.95 mg, 1.0 mg, 2.0 mg, 3.0 mg, 4.0 mg, 5.0 mg, 10.0 mg, 15.0 mg, 20.0 mg, or 25.0 mg, or may fall within a range bounded by any two of the foregoing values.For example, in certain embodiments, a sustained release formulation (e.g., an ocular implant) may be exactly or approximately 0.1 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.35 mg, 0.4 mg, 0.45 mg, 0.5 mg, 0.55 mg, 0.6 mg, 0.65 mg, 0.7 mg, 0.75 mg, 0.8 mg, 0.85 mg, 0.9 mg, 0.95 mg, 1.0 mg, 2.0 mg, 3.0 mg, 4.0 mg, 5.0 mg, 10.0 mg, 15.0 mg, 20.0 mg, or 25.0 mg of anti-CFHR4 antibody, or an amount falling within a range bounded by any two of the foregoing values.

[0133] Therapeutic or prophylactic efficacy can be monitored by periodic evaluation of the treated patient. In the case of repeated administration over several days or more, depending on the condition, the treatment is repeated until the desired suppression of disease symptoms occurs. However, other dosage regimes may be useful and are within the scope of the present disclosure. The desired dosage can be delivered by a single bolus administration of the composition, by multiple bolus administrations of the composition, or by continuous infusion administration of the composition. The composition comprising the anti-CFHR4 antibody, or an antigen-binding fragment thereof, can be administered to a mammal using standard administration techniques, including ocular, oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. The composition is preferably suitable for ocular administration.

[0134] In accordance with the compositions and methods of treatment described herein, embodiments of the present disclosure include anti-CFHR4 antibodies with enhanced half-life (e.g., following ocular administration), such that the anti-CFHR4 antibodies can be administered less frequently to a subject. In some embodiments, the antibody comprises a half-life extending moiety. In some embodiments, the half-life extending moiety comprises a polypeptide that can be attached to an anti-CFHR4 antibody of the present disclosure by any means known in the art (e.g., generating a fusion protein). In some embodiments, the polypeptide that can be attached to an anti-CFHR4 antibody of the present disclosure is at least 20 amino acids in length and comprises any combination of G, A, ST, E, and P residues. In some embodiments, the half-life extending polypeptide is attached to the C-terminus or N-terminus of the antibody. In some embodiments, this is referred to as "Xtenylation," as further described in US8933197, US7846445, US7855279, US8492530, US9938331, US8673860, US9371369, US9926351, US10961287, US10172953, and US10953073.

[0135] The present disclosure also provides compositions comprising any of the anti-CFHR4 antibodies or antigen-binding fragments thereof described herein. The compositions are desirably pharma- ceutically acceptable (e.g., physiologically acceptable) compositions comprising a carrier, preferably a pharma- ceutically acceptable (e.g., physiologically acceptable) carrier, and a therapeutic agent and / or an anti-CFHR4 antibody or antigen-binding fragment thereof. Any suitable carrier may be used within the context of the present disclosure, and such carriers are well known in the art. For example, the compositions may include a preservative, such as, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. Optionally, a mixture of two or more preservatives may be used. Additionally, a buffering agent may be included in the composition. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. Optionally, a mixture of two or more buffering agents may be used. Methods for preparing compositions for pharmaceutical use are known to those skilled in the art and are described, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0136] After administration to a mammal (e.g., a human), the biological activity of an anti-CFHR4 antibody, or an antigen-binding fragment thereof, can be measured by any suitable method known in the art. For example, biological activity can be assessed by determining the stability of the anti-CFHR4 antibody. The biological activity of an anti-CFHR4 antibody can also be assessed by determining its binding affinity to a CFHR4 peptide and / or by assessing its binding affinity to a peptide that may cross-react with it. The term "affinity" refers to the equilibrium constant for the reversible binding of two agents and is defined as the dissociation constant (K D) The affinity of a binder for a ligand, e.g., the affinity of an antibody for an epitope, can range, for example, from about 1 femtomolar (fM) to about 1 millimolar (mM) (e.g., from about 1 picomolar (pM) to about 1 nanomolar (nM), or from about 1 nM to about 1 micromolar (μM)). In some embodiments, the affinity of an anti-CFHR4 antibody can be from about 1 nm to about 20 nm, desirably from about 5 nm to about 10 nm. Antibody affinity for an antigen or epitope of interest can be measured using any art-recognized assay. Such methods include, for example, fluorescence-activated cell sorting (FACS), separable beads (e.g., magnetic beads), antigen panning, and / or ELISA (see, e.g., Janeway et al. (eds.), Immunobiology, 5th ed., Garland Publishing, New York, NY, 2001).

[0137] In some embodiments, the anti-CFHR4 antibody or a composition comprising the anti-CFHR4 antibody may be administered alone or in combination with other drugs. For example, the anti-CFHR4 antibody may be administered in combination with other drugs for the treatment or prevention of AMD and / or GA as disclosed herein. For example, the anti-CFHR4 antibody of the present disclosure, or an antibody conjugate, fusion protein, or polymeric formulation thereof, may be used in therapy alone or in combination with other drugs. For example, the anti-CFHR4 antibody of the present invention may be co-administered with at least one additional therapeutic agent. In certain embodiments, the additional therapeutic agent is another antibody, a chemotherapeutic agent, a cytotoxic agent, an antiangiogenic agent, an immunosuppressant, a prodrug, a cytokine, a cytokine antagonist, a cytotoxic radiation therapy, a corticosteroid, an antiemetic agent, a cancer vaccine, an analgesic agent, or a growth inhibitory agent, or a combination thereof.

[0138] For example, in certain embodiments, any of the preceding methods further comprises administering one or more additional compounds. In certain embodiments, the anti-CFHR4 antibody, antibody conjugate, fusion protein, or polymer formulation is administered simultaneously with the additional compound(s). In certain embodiments, the anti-CFHR4 antibody, antibody conjugate, fusion protein, or polymer formulation is administered before or after the additional compound(s). In certain embodiments, the additional compound(s) bind to a second biological molecule selected from the group consisting of VEGF, IL-1β; IL-6; IL-6R; IL-13; IL-13R; PDGF; angiopoietin; Ang2; Tie2; S1P; integrins αvβ3, αvβ5, and α5β1; betacellulin; apelin / APJ; erythropoietin; complement factor D; TNFα; HtrA1; VEGF receptor; ST-2 receptor; and proteins genetically related to AMD risk, e.g., complement pathway components C2, factor B, factor H, CFHR3, C3b, C5, C5a, and C3a; HtrA1; ARMS2; TIMP3; HLA; interleukin-8 (IL-8); CX3CR1; TLR3; TLR4; CETP; LIPC; COL10A1; and TNFRSF10A. In some embodiments, the additional compound is an antibody or an antigen-binding fragment thereof. In some embodiments according to any of the above embodiments (or as applied to any of the above embodiments), the ocular disorder is an intraocular neovascular disease selected from the group consisting of proliferative retinopathy, choroidal neoplasia (CNV), age-related macular degeneration (AMD), geographic atrophy (GA), diabetic and other ischemia-related retinopathies, diabetic macular edema, pathological myopia, von Hippel-Lindau disease, ocular histoplasmosis, retinal vein occlusion (RVO) (including CRVO and BRVO), corneal neovascularization, retinal neovascularization, and retinopathy of prematurity (ROP).

[0139] In some cases, the anti-CFHR4 antibodies, or antibody conjugates, fusion proteins, and / or polymeric formulations thereof of the present disclosure may be administered in combination with at least one additional therapeutic agent for the treatment of an ocular disorder described herein (e.g., AMD (e.g., wet AMD or dry AMD), GA, DME, DR, or RVO).Exemplary additional therapeutic agents for combination therapy for the treatment of ocular disorders include, for example, antiangiogenic agents such as VEGF antagonists, including anti-VEGF antibodies (e.g., anti-VEGF Fab LUCENTIS® (ranibizumab)), soluble receptor fusion proteins (e.g., recombinant soluble receptor fusion protein EYLEA® (aflibercept, also known as VEGF Trap Eye, Regeneron / Aventis)), aptamers (e.g., anti-VEGF pegylated aptamer MACUGEN® (pegaptanib sodium, NeXstar Pharmaceuticals / OSI Pharmaceuticals), and VEGFR tyrosine kinase inhibitors (e.g., 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474), 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline (AZD2171), vatalanib (PTK787), semaxaminib (SU5416, SUGEN), and SUTENT® (sunitinib)); tryptophanyl-tR NA synthetase (TrpRS); squalamine; RETAANE® (anecoltave acetate for depot suspension, Alcon, Inc.); combretastatin A4 prodrug (CA4P); MIFEPREX® (mifepristone-ru486); subtenon triamcinolone acetonide; intravitreal crystalline triamcinolone acetonide; matrix metalloproteinase inhibitors (e.g., Prinomast (AG3340, Pfizer)); fluocinolone acetonide (including fluocinolone intraocular implant, Bausch & Lomb / Control Delivery Systems); linomide; inhibitors of integrin β3 function; angiostatin, and combinations thereof.

[0140] Further examples of additional therapeutic agents that can be used in combination with the anti-CFHR4 antibodies, or antibody conjugates, fusion proteins, and / or polymeric formulations thereof of the present disclosure for the treatment of ocular disorders (e.g., AMD, GA, DME, DR, or RVO) include VISUDYNE® (verteporfin, a light-activated drug typically used in combination with photodynamic therapy using non-thermal lasers), PKC412, Endovion (NS 3728, NeuroSearch A / S), neurotrophic factors (e.g., glial-derived neurotrophic factor (GDNF) and ciliary neurotrophic factor (CNTF)), diltiazem, dorzolamide, PHOTOTROP®, 9-cis-retinal, ophthalmic drugs (e.g., phosphodiesterase, echothiophate, or carbonic anhydrase inhibitors), beovastat (AE-941, AEterna Laboratories, Inc.), Sirna-027 (AGF-745, Sirna Therapeutics, Inc.), neurotrophins (including, by way of example only, NT-4 / 5, Genentech), Cand5 (Acuity Pharmaceuticals), INS-37217 (Inspire Pharmaceuticals), integrin antagonists (including those from Jerini AG and Abbott Laboratories), EG-3306 (Ark Therapeutics Ltd.), BDM-E (BioDiem Ltd.), thalidomide (where used, e.g., EntreMed, Inc.), cardiotrophin-1 (Genentech), 2-methoxyestradiol (Allergan / Oculex), DL-8234 (Toray Industries), NTC-200 (Neurotech), tetrathiomolybdate (University of Michigan), LYN-002 (Lynkeus Biotech), microalgae compounds (Aquasearch / Albany, Mera Pharmaceuticals), D-9120 (Celitech Group plc), ATX-S10 (Hamamatsu Photonics), TGF-beta 2 (Genzyme / Celtrix), tyrosine kinase inhibitors (e.g., from Allergan, SUGEN, or Pfizer), NX-278-L (NeXstar Pharmaceuticals / Gilead Sciences), Opt-24 (OPTIS France SA), retinal ganglion neuroprotectants (Cogent Neurosciences), N-nitropyrazole derivatives (Texas A&M University System), KP-102 (Krenitsky Pharmaceuticals), cyclosporine A, therapeutic agents used in photodynamic therapy (e.g., VISUDYNE®; receptor-targeted PDT, Bristol-Myers Squibb, Co.; porfimer sodium for injection with PDT; verteporfin, QLT Inc.; rostaporfin with PDT, Miravent Medical Technologies; talaporfin sodium with PDT, Nippon Petroleum; and motexafin lutetium (Pharmacyclics, Inc.)), antisense oligonucleotides (examples include products tested by Novagali Pharma SA and ISIS-13650, Isis Pharmaceuticals), and combinations thereof.

[0141] The anti-CFHR4 antibodies, or antibody conjugates, fusion proteins, and / or polymeric formulations thereof of the disclosure may be used in therapeutic or surgical procedures to treat ocular disorders (e.g., AMD, GA DME, DR, or RVO), such as laser photocoagulation (e.g., panretinal photocoagulation (PRP)), drusen laser therapy, macular hole surgery, macular relocation surgery, implantable miniature telescopes, PHI motion angiography (also known as microlaser therapy and feeder vessel therapy), proton therapy, microstimulation therapy, retinal detachment and vitreous surgery, scleral buckle, submacular surgery, transpupillary thermotherapy, photosystem I therapy, use of RNA interference (RNAi), extracorporeal rheopheresis (also known as membrane fractionation and rheotherapy), microchip implantation, stem cell therapy, gene replacement therapy, ribozyme gene therapy (including hypoxia response element gene therapy, Oxford It may also be administered in combination with erythropoietin, Biomedica; Lentipak, Genetix; and PDEF gene therapy, GenVec), photoreceptor / retinal cell transplants (including transplantable retinal epithelial cells, Diacrin, Inc., retinal cell transplants, Cell Genesys, Inc.), acupuncture, and combinations thereof.

[0142] In some embodiments, the anti-CFHR4 antibodies, or antibody conjugates, fusion proteins, and / or polymeric formulations thereof of the present disclosure may be administered in combination with an anti-angiogenic agent for the treatment of ocular disorders (e.g., AMD, GA, DME, DR, or RVO). Any suitable anti-angiogenic agent may be used in combination with the antibodies of the present disclosure, including, but not limited to, those listed by Carmeliet et al. Nature 407:249-257,2000. In some embodiments, the anti-angiogenic agent is a VEGF antagonist including an anti-VEGF antibody (e.g., anti-VEGF Fab LUCENTIS® (ranibizumab), RTH-258 (formerly ESBA-1008, an anti-VEGF single chain antibody fragment, Novartis), or a bispecific anti-VEGF antibody (e.g., an anti-VEGF / anti-Angiopoietin 2 bispecific antibody, e.g., RG-7716, Roche)), a soluble recombinant receptor fusion protein (e.g., EYLEA® (aflibercept)), a VEGF variant, a soluble VEGFR fragment, an aptamer capable of blocking VEGF (e.g., pegaptanib) or VEGFR, a neutralizing anti-VEGFR antibody, a small molecule inhibitor of VEGFR tyrosine kinase, an anti-VEGF DARPins® (e.g., abicipar pegol), small interfering RNA that inhibits the expression of VEGF or VEGFR, VEGFR tyrosine kinase inhibitors (e.g., 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474), 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline (AZD2171), vatalanib (PTK787), semaxaminib (SU5416, SUGEN), and SUTENT® (sunitinib)), and combinations thereof.

[0143] Other suitable anti-angiogenic agents that may be administered in combination with the antibodies, or antibody conjugates, fusion proteins, and / or polymeric formulations thereof of the present disclosure for the treatment of ocular disorders (e.g., AMD, GA, DME, DR, or RVO) include corticosteroids, angiostatic steroids, anecoltate acetate, angiostatin, endostatin, tyrosine kinase inhibitors, matrix metalloproteinase (MMP) inhibitors, insulin-like growth factor binding protein 3 (IGFBP3), interleukin-1 (IL-1) inhibitors, IL-1 ... These include stromal derived factor (SDF-1) antagonists (e.g., anti-SDF-1 antibodies), pigment epithelium derived factor (PEDF), gamma-secretase, delta-like ligand 4, integrin antagonists, hypoxia inducible factor (HIF)-1α antagonists, protein kinase CK2 antagonists, agents that inhibit stem cells (e.g., endothelial progenitor cells) homing to sites of angiogenesis (e.g., anti-vascular endothelial cadherin (CD-144) antibodies and / or anti-SDF-1 antibodies), and combinations thereof.

[0144] In some embodiments, the anti-CFHR4 antibodies, or antibody conjugates, fusion proteins, and / or polymeric formulations thereof of the present disclosure may be administered in combination with agents active against angiogenesis for the treatment of ocular disorders (e.g., AMD, GA, DME, DR, or RVO), such as anti-inflammatory agents, mammalian target of rapamycin (mTOR) inhibitors (e.g., rapamycin, AFINITOR® (everolimus)), and TORISEL® (temsirolimus), cyclosporine, tumor necrosis factor (TNF) antagonists (e.g., anti-TNFα antibodies or antigen-binding fragments thereof (e.g., infliximab, adalimumab, certolizumab pegol, and golimumab), or soluble receptor fusion proteins (e.g., etanercept)), anti-complement agents, nonsteroidal anti-inflammatory agents (NSAIDs), or combinations thereof.

[0145] Any suitable AMD therapeutic may be administered as an additional therapeutic agent in combination with the anti-CFHR4 antibodies, or antibody conjugates, fusion proteins, and / or polymer formulations thereof of the present disclosure for the treatment of an ocular disorder (e.g., AMD, GA, DME, DR, or RVO), including, but not limited to, a VEGF antagonist, such as an anti-VEGF antibody (e.g., LUCENTIS® (ranibizumab), RTH-258 (formerly ESBA-1008, an anti-VEGF single chain antibody fragment, Novartis), or a bispecific anti-VEGF antibody (e.g., an anti-VEGF / anti-Angiopoietin 2 bispecific antibody, e.g., RG-7716, Roche)), a soluble VEGF receptor fusion protein (e.g., EYLEA® (aflibercept)), an anti-VEGF DARPin® (e.g., abicipar pegol, Molecular Partners), AG / Allergan), or anti-VEGF aptamers (e.g., MACUGEN® (pegaptanib sodium)), platelet-derived growth factor (PDGF) antagonists, such as anti-PDGF antibodies, anti-PDGFR antibodies (e.g., REGN2176-3), anti-PDGF-BB pegylated aptamers (e.g., FOVISTA®, Ophthotech / Novartis), soluble PDGFR receptor fusion proteins, or PDGF / VEGF dual antagonists (e.g., small molecule inhibitors (e.g., DE-120 (Santen) or X-82 (TyrogeneX)), or bispecific anti-PDGF / anti-VEGF antibodies )), VISUDYNE® (verteporfin) in combination with photodynamic therapy, antioxidants, complement system antagonists, e.g., complement factor C5 antagonists (e.g., small molecule inhibitors (e.g., ARC-1905, Opthotech), or anti-C5 antibodies (e.g., LFG-316, Novartis), properdin antagonists (e.g., anti-properdin antibodies, e.g., CLG-561, Alcon), or complement factor D antagonists (e.g., anti-complement factor D antibodies, e.g., lampalizumab, Roche)), visual cycle regulators (e.g., emixustat hydrochloride), and squalamine (e.g., OHR-102, OhrPharmaceutical), vitamin and mineral supplements (e.g., those described in Age-Related Eye Disease Study 1 (AREDS1, zinc and / or antioxidants) and Study 2 (AREDS2, zinc, antioxidants, lutein, zeaxanthin, and / or omega-3 fatty acids)), cell-based therapies, e.g., NT-501 (Renexus), PH-05206388 (Pfizer), huCNS-SC cell transplantation (StemCells), CNTO-2476 (Janssen), OpRegen (Cell Cure Neurosciences), or MA09-hRPE cell transplantation (Ocata Therapeutics), tissue factor antagonists (e.g., hl-con1, Iconic Therapeutics), alpha-adrenergic receptor agonists (e.g., brimonidine tartrate), peptide vaccines (e.g., S-646240, Shionogi), amyloid beta antagonists (e.g., anti-beta amyloid monoclonal antibodies, e.g., GSK-933776), S1P antagonists (e.g., anti-S1P antibodies, e.g., iSONEP™, Lpath Inc), ROBO4 antagonists (e.g., anti-ROBO4 antibodies, e.g., DS-7080a, Daiichi Sankyo), lentiviral vectors expressing endostatin and angiostatin (e.g., RetinoStat), and any combination thereof. In some cases, the AMD therapeutics (including any of the preceding AMD therapeutics) can be co-formulated. For example, the anti-PDGFR antibody REGN2176-3 can be co-formulated with aflibercept (EYLEA®). In some cases, such co-formulations may be administered in combination with an antibody of the present disclosure. In some cases, the ocular disorder is AMD (e.g., wet AMD).

[0146] In addition to therapeutic uses, the anti-CFHR4 antibodies or antigen-binding fragments described herein may be used for diagnostic or research applications. Research applications include, for example, methods that utilize anti-CFHR4 antibodies and a label to detect CFHR4 in a sample (e.g., in a human body fluid or in a cell or tissue extract). Anti-CFHR4 antibodies or antigen-binding fragments thereof may be used in any suitable assay for measuring CFHR4 in a sample for diagnostic and / or research purposes. Such assays include, but are not limited to, sandwich immunoassays, enzyme immunoassays (EIA), enzyme-linked immunosorbent assays (ELISA), lateral flow assays, competitive inhibition immunoassays (e.g., forward and reverse), competitive binding assays, Forster resonance energy transfer (FRET), one-step antibody detection assays, single molecule detection assays, radioimmunoassays (RIA), and FACS. Such methods are disclosed, for example, in U.S. Pat. Nos. 6,143,576, 6,113,855, 6,019,944, 5,985,579, 5,947,124, 5,939,272, 5,922,615, 5,885,527, 5,851,776, 5,824,799, 5,679,526, 5,525,524, and 5,480,792, and in Adamczyk et al., Anal. Chim. Acta, 579(1):61-67 (2006).

[0147] The anti-CFHR4 antibody or antigen-binding fragment thereof may be provided in a kit, e.g., packaged with predetermined amounts of reagents and instructions for using the antibody to perform an assay (e.g., an assay to detect CFHR4). Thus, the present disclosure provides a kit comprising an antibody or antigen-binding fragment described herein and instructions for its use. The instructions may be in either paper form or in computer-readable form, such as a disk, CD, DVD, etc. Alternatively or additionally, the kit may include calibrators or controls, and / or at least one container (e.g., tube, microtiter plate, or strip) for performing the assay, and / or a buffer, such as an assay buffer or wash buffer. Ideally, the kit includes all components necessary to perform the assay, i.e., reagents, standards, buffers, diluents, etc. Additionally, other additives, such as stabilizers, buffers (e.g., blocking buffer or lysis buffer), etc., may be included in the kit. The relative amounts of the various reagents may be widely varied to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay. Reagents may be provided as dry powders (usually lyophilized) that include excipients that, upon dissolution, provide a reagent solution having the appropriate concentration.

[0148] The following examples further illustrate various embodiments of the present disclosure but, of course, should not be construed as in any way limiting its scope. EXAMPLES

[0149] It will be appreciated by those skilled in the art that other suitable modifications and adaptations of the disclosed methods described herein are readily applicable and recognizable, and may be made using suitable equivalents without departing from the scope of the disclosure or the aspects and embodiments disclosed herein. Having described the disclosure in detail, the disclosure will be more clearly understood by reference to the following examples, which are intended merely to illustrate some aspects and embodiments of the disclosure, and should not be considered as limiting the scope of the disclosure. The disclosures of all journal references, U.S. patents, and publications referenced herein are incorporated herein by reference in their entirety.

[0150] The present disclosure has multiple aspects, which are illustrated by the following non-limiting examples.

[0151] Example 1 Genetic analysis of CFHR4 in geographic atrophy. This example describes the analysis carried out to identify the effect of CFHR1 / 4 deletion on age-related macular degeneration (AMD) in FinnGen. There are three common haplotypes consisting of a variant of CFH (rs1061170 Y402H) and a variant that is an eQTL and pQTL of CFH and CFHR4 (rs10922109), which are associated with three different levels of risk. The high AMD risk haplotype (H402+high CFHR4 expression, rs106117_C(ref) / rs10922109_C(ref)) was set as the "reference" haplotype to allow comparison with other haplotypes (frequency in cases: 0.56, frequency in controls: 0.36). Carrying Y402 (alternative allele at rs106117) and high CFHR4 / low CFH expression (reference allele at rs10922109) is associated with a "medium" AMD risk (frequency in cases: 0.196, frequency in controls: 0.21). A "low" risk haplotype carries Y402 (alternative allele at rs106117) and has low CFHR4 / high CFH expression (alternative allele at rs10922109), frequency in cases: 0.22, frequency in controls: 0.43). It was hypothesized that haplotypes carrying CFHR1 / 4 deletions have a different risk of AMD in a medium-risk background.

[0152] Genetic variation at this locus provides a natural experiment to determine the impact of the CFHR1 / 4 deletion on AMD. Although the CFHR1 / 4 deletion has not been directly genotyped in FinnGen, there are variants (rs528922402 and rs188297593) that incompletely tag (r2=0.36 in Finland, D'=1) a non-overlapping subset of the deletion in the Finnish population of the 1000 Genomes dataset (HapMap Phase 3). Variant rs188297593 has a D'<1.0 in HapMap Phase 3 samples of European ancestry (1 in 4 carriers does not tag the CFHR1 / 4 deletion). These two variants capture a subset but not all carriers of the deletion.

[0153] AMD risk in individuals from FinnGen R6 was examined to determine the effect of CFH variants (rs1061170), CFHR4 / CFH eQTL / pQTL (rs10922109) and CFHR1 / 4 deletions (tagged by rs528922402 and rs188297593). Haplotypes composed of individual variants were generated and logistic regression was performed in R to predict the effect of haplotypes on AMD risk adjusted for sex, age and PC. The reference haplotype was used as a reference to which the effects of all other haplotypes were correlated.

[0154] Consistent with the hypothesis that CFHR4 directly contributes to AMD risk, a 'medium' risk haplotype with a CFHR4 deletion conferred increased protection from AMD compared with a 'medium' risk haplotype with an intact CFHR4.

[0155] [Table 1]

[0156] Example 2 Generation of recombinant CFHR4 proteins. Recombinant protein preparations were undertaken to generate specific proteins that could be used as immunogens / antigens, screening reagents, and / or control reagents. All efforts were directed toward the goal of generating a panel of anti-CFHR4 antibodies with desired properties.

[0157] Proteins with sequences corresponding to human CFHR4b, human CFHL-1 wild type, human CFHL-1 with tyrosine 402 mutated to histidine, and cynomolgus CFHR4b were successfully produced using various protein tags. At completion, all protein preparations were greater than 90% pure by analytical methods and contained less than 1 endotoxin unit per milliliter. Examples of the final specifications of the individual preparations are shown below.

[0158] Starting with in silico analysis, appropriate wild-type amino acid sequences of human and cynomolgus CFHR4b and human CFHL-1 were identified and extracted from public databases. The sequences were then further analyzed and manually modified using Geneious Prime software according to the experimental design. The sequences were tested for susceptibility and modified to add additional amino acid sequences encoding "tags" to facilitate purification, reduce immunogenicity, or simplify analytical screening. Examples of protein tags used in this study include the HIS tag, the unique mouse IgG Fc tag, and the Avi tag. The tags were separated from each other and from the CFHR4b or CFHL-1 sequences with short linker sequences. All tags were added to the C-terminus of the sequences. Additionally, the native signal peptide was removed and replaced with a signal peptide that facilitates recombinant expression.

[0159] After all modifications were completed, the amino acid sequences were reverse translated into DNA sequences and optimized for codon bias found in the human genome. These optimized DNA sequences were sent to Integrated DNA Technologies and generated as DNA fragment(s) with DNA overhangs added to the 5' and 3' ends. Using DNA overhangs and the Gibson cloning method, these DNA fragments were assembled into the expected sequences and cloned into a mammalian expression plasmid driven by a CMV promoter. The plasmids were grown in E. coli with appropriate antibiotic selection and prepared at a scale useful for recombinant expression using a commercial preparation kit purchased from Qiagen. The sequences of the plasmids and expressed genes were then confirmed using Sanger sequencing.

[0160] The sequence verified plasmid was transfected into human embryonic kidney cells adapted for recombinant expression using polyethyleneimine. One day after transfection, the cells were supplemented with chemicals and nutrients designed to increase recombinant protein expression. These supplements included sodium propionate, valproic acid, glucose, glutamine, and various yeast lysates. Five days after transfection, the expressing cell cultures were harvested. As the recombinant protein was secreted into the growth medium, cells, cell fragments, and cell debris were removed by centrifugation and filtration through membranes with pores of 0.22 microns or less. The clarified medium conditioned with the recombinant protein was now ready for purification.

[0161] Proteins of interest were purified from cell culture media using FPLC (fast pressurized liquid chromatography) and appropriate commercially available pre-packed affinity chromatography columns for the C-terminal tag(s) and immobilized on a chromatography column. For HIS-tagged proteins, a Ni-NTA agarose column was used. For Fc-tagged proteins, a proA agarose column was used. Multiple column washes followed, each specific to the column / tag / chromatography type. For HIS-tagged proteins, the protein of interest was eluted from the column with 300 mM imidazole, and for Fc-tagged proteins, it was eluted from the column with 100 mM citrate pH 3.5. After elution, Fc-tagged proteins were adjusted to neutral pH using 1 M Hepes pH 9. For HIS-tagged proteins, no neutralization was required as the elution buffer was at neutral pH.

[0162] Protein quality and quantity were assessed using a combination of SDS-PAGE gel electrophoresis, spectrophotometry, and analytical SEC (size-exclusion chromatography). Most proteins for this project required further purification. FPLC and a second chromatography column were used to further purify the proteins. Separation by size achieved with the size-exclusion column allowed for an increase in protein purity to >90%. SEC also allowed for buffer exchange from the affinity chromatography elution buffer to the final selection buffer (PBS). Quality control of the protein samples was then moved on. If the protein concentration needed to be increased at any point during purification, the protein was concentrated by separating it from the buffer using an Amicon Ultra molecular weight cut-off (MWCO) filtration unit. The MWCO was chosen to ensure compatibility with the size of the protein of interest. All final samples were concentrated to greater than 1 milligram per milliliter before moving on to final quality control.

[0163] The final protein samples were assayed for endotoxin contamination using Charles River's EndoSafe PTS system. Final protein concentration was determined by spectrophotometry. Three micrograms of the final sample was injected onto an analytical SEC column (YMC Diol 300) to determine its final purity. Additionally, SDS-PAGE electrophoresis was performed in some cases to determine final quality. Once all final metrics were passed, the protein was sterilized using a sterile 0.22 micron filter in a biosafety cabinet. This was followed by aseptic aliquoting and flash freezing in liquid nitrogen before storage at -80°C.

[0164] Example 3 Recovery of CFHR4 antibody sequences from immunized mice. CFHR4 Immunization: Three cohorts of Alloy Therapeutic transgenic humanized mice (ATX-GK) were immunized with human CFHR4b using the following 5-week protocol (Figures 3A-3C):

[0165] Cohort 1: Five ATX-GK mice immunized with human CFHR4B (ATX-P-57) using a standard 5-week RIMMS protocol of 10ug subcutaneous administration of antigen emulsified in complete Freund's adjuvant followed by five weekly subcutaneous administrations of antigen emulsified in incomplete Freund's adjuvant. Cohort 2: Five ATX-GK mice immunized with human CFHR4B (ATX-P-57) using a 5-week RIMMS protocol of 10ug subcutaneous administration of antigen emulsified in incomplete Freund's adjuvant. Cohort 3: Five ATX-GK mice immunized with human CFHR4B (ATX-P-57) using a 5-week RIBI IP protocol of 10ug intraperitoneal administration of antigen emulsified in RIBI adjuvant.

[0166] Samples were drawn at week 4 and tested by ELISA for antigen positive and purification tag negative serum titers. ELISA plates were coated with either 1ug / ml of CFHR4 immunogen or an irrelevant protein (P114) with the same purification tag as the immunogen. Antigen-coated plates were incubated with seven 10-fold serial dilutions of serum starting at 1:300. Antibody bound to antigen was detected by anti-mouse IgG HRP secondary antibody and one-step TMB solution. Absorbance signal at 450nm was measured with an ELISA microplate reader.

[0167] Hybridomas: Immune tissues from high titer mice were harvested and stored for antibody discovery. Hybridoma cell lines producing CFHR4 antibodies were generated by fusion of single B cells from the spleen and lymph nodes of titer positive mice with myeloma cells. Twenty 96-well plates of hybridoma fusions were generated and grown. Hybridomas expressing CFHR4 specific antibodies were detected by antigen binding by ELISA. The affinity of the antibodies in the hybridoma supernatants was measured by SPR using an Octet instrument. CFHR4 antibodies in the hybridoma supernatants were loaded onto a biosensor. The response was measured as a nm shift in the interference pattern and was proportional to the number of antibodies bound to the biosensor surface. The binding interaction of CFHR4 to the immobilized antibodies was measured as association (kon). After analyte association, the biosensor was immersed in PBS without CFHR4 to dissociate the bound antigen from the antibodies (kdis). The KD(M), or affinity, of the antibodies for CFHR4 was measured as kdis / kon. The heavy and light chains from confirmed hybridomas were sequenced. RNA was isolated from CFHR4 antibody secreting hybridomas and the heavy and light chain variable regions were cloned by reverse transcription using gene-specific primers followed by PCR amplification using variable chain gene-specific primers. PCR products were sequenced by standard Sanger sequencing methods.

[0168] Phage display: Variable heavy and light chains were amplified from the spleens of high titer immunized mice by reverse transcription using gene-specific primers followed by PCR amplification using variable chain gene-specific primers. The variable regions were cloned into a phage display vector designed to express Fabs in the phage g3p protein. A library of phages expressing unique Fabs was amplified and purified. Phages were bound to biotinylated CFHR4 antigen captured on streptavidin magnetic beads. Phages that remained bound to the antigen beads after several stringent washes were eluted using a basic triethylamine solution and neutralized with Tris buffer pH 8.0. Eluted phages were reinfected into TG1 bacterial cells, amplified by co-infection with M13 helper phage, and purified by PEG precipitation. Purified phages expressing Fabs were selected for antigen binding as described. Phages from the second round were diluted and infected into TG1 cells. Polyclonal pools of phage released from two rounds of panning were tested by ELISA to confirm that the pools contained CFHR4-specific phage. Variable heavy and light chain regions were sequenced from single infected bacterial colonies using rolling circle amplification and standard Sanger sequencing.

[0169] Antibody Sequencing: Unique variable heavy and light chain pairs from the hybridoma and phage display campaigns were cloned into vectors designed to express full-length antibodies as IgG in HEK293 cells under the control of a CMV promoter. Antibody expression vectors were complexed with polyethylenimine and transfected into HEK293 cultures. After 5 days of shaking at 37°C in 293 cell culture medium, antibodies were captured on an agarose-based Protein A resin. After several stringent washes, antibodies were eluted in a glycine solution (pH 3), neutralized with Hepes (pH 9), and buffer exchanged into PBS.

[0170] Example 4 Human CFHR4 monoclonal antibody differential scanning fluorimetry (DSF). The development of effective monoclonal antibodies depends not only on their biological activity but also on their physicochemical properties such as homogeneity and stability. The stability of mAbs can be affected by their formulation. Among the many techniques used to examine mAb stability, differential scanning fluorimetry (DSF) offers both excellent throughput and minimal material consumption. DSF measures the temperature (Tm) of protein unfolding transition based on the change in fluorescence intensity of an environmentally sensitive dye.

[0171] Experiments were performed to evaluate the thermal stability of the human CFHR4 monoclonal antibody of the present disclosure (the "ATX" antibody) by determining the melting temperature. Thermal stability was evaluated by differential scanning fluorimetry (DSF) utilizing the Protein Thermal Shift (PTS) assay from Applied Biosystems. The assay was performed according to the manufacturer's instructions. Briefly, the antibody to be evaluated was prepared in triplicate by mixing with a protein thermal shift dye and buffer. Real-time melting experiments were performed from 25°C to 95°C on a QuantStudio 3. Data was analyzed by using Protein Thermal Shift software, and melting temperatures (Tm) were calculated from the melting curves (Figure 4).

[0172] [Table 2]

[0173] Example 5 Cross-blocking of CFHR4 antibodies. High-throughput epitope binning experiments were performed on a real-time label-free biosensor (Carterra LSA) to sort a large panel of mAbs into bins based on their ability to block each other for binding to the antigen. In pairwise epitope binning analysis, antigen and antibody 2 (analyte antibody) are sequentially applied to a sensor chip (HC200M) pre-loaded with antibody 1 (ligand antibody) covalently linked to the antigen. An increase in response upon exposure to the analyte antibody indicates non-competition between the two antibodies, while a lack of change in signal indicates competition. Antibodies with the same blocking profile relative to others in the test set are grouped into one bin. Community network plots are used to explore the clustering of mAbs that share similar, but not necessarily identical, competition profiles. Rather than strictly relying on sandwich / blocking assignments in the heatmaps, as in the bin network plots (Figure 5A), hierarchical clustering is applied to the sorted heatmaps to generate network plots (Figure 5B), which group mAbs successively.

[0174] Example 6 CFHR4 antibody binding kinetics. Kinetics experiments were performed on a Carterra LSA with a running buffer of PBS pH 7.40, 1% BSA, 0.05% Tween 20. Antibodies were covalently bound and printed onto HC30M chips. The chips were activated with 33 mM s-NHS and 133 mM EDC in 100 mM MES pH 5.5 for 7 min. Antibodies at 10 mg / ml in acetate buffer pH 4.5 were used for printing for 10 min. The printed chips were then quenched with 1 M ethanolamine pH 8.5 for 7 min. For kinetic analysis, purified recombinant his-tagged proteins ATX-P-57, human CDFR4B-His at concentrations from 0.076 nM to 1500 nM (serial 3-fold dilutions) were injected sequentially. For each concentration, 5 min of association was performed followed by 15 min of dissociation. Results were processed and analyzed with Carterra LSA Kinetics Software. Kinetic data were referenced to an intervening reference spot, double referenced to buffer cycles, and then globally fitted to a 1:1 binding model to determine their apparent association and dissociation rate constants (ka and kd values). The ratio of kd / ka was used to derive the KD value for each antigen / mAb interaction, i.e., KD=kd / ka (Figure 6).

[0175] Example 7 Cross-reactivity of CFHR4 antibodies. Experiments were performed to determine the reactivity of the disclosed CFHR4 antibodies (developed using the CFHR4b antigen, see Example 3) with various other CFHR4 proteins. Experiments were performed to determine the reactivity of CFHR4 antibodies with cynomolgus monkey CFHR4b protein (cCFHR4b), as shown in FIG. 7. Binding experiments were performed on a Carterra LSA with a running buffer of PBS pH 7.40, 1% BSA, 0.05% Tween 20. Antibodies were covalently bound and printed on HC30M chips. The chips were activated with 33 mM s-NHS and 133 mM EDC in 100 mM MES pH 5.5 for 7 minutes. 10 mg / ml of antibody in acetate buffer pH 4.5 was used for printing for 10 minutes. The printed chips were then quenched with 1 M ethanolamine pH 8.5 for 7 minutes. In the reactivity assay, 160 nM ATX-P-141, cyno CFHR4B-mFc was tested with 5 min association followed by 15 min dissociation. Results were processed and analyzed with Carterra LSA Kinetics Software. Data was referenced to an intervening reference spot, double referenced to buffer cycles, and then responses (nm) after association were reported. Isotype controls were used to determine the cutoff response for positive binding.

[0176] Experiments were also performed to determine the reactivity of CFHR4 antibodies with human CFHR4a protein (Figure 8). Binding experiments were performed in a Carterra LSA with a running buffer of PBS pH 7.40, 1% BSA, 0.05% Tween 20. Antibodies were covalently bound and printed onto HC30M chips. The chips were activated with 33 mM s-NHS and 133 mM EDC in 100 mM MES pH 5.5 for 7 minutes. 10 mg / ml of antibody in acetate buffer pH 4.5 was used for printing for 10 minutes. The printed chips were then quenched with 1 M ethanolamine pH 8.5 for 7 minutes. In the reactivity assay, 540 nM ATX-P-56, human CFHL4A-His was tested with 5 minutes of association followed by 15 minutes of dissociation. Results were processed and analyzed with Carterra LSA Kinetics Software. Data was referenced to an intervening reference spot, double referenced to buffer cycles, and then responses (nm) after association were reported. Isotype controls were used to determine the cutoff response for positive binding.

[0177] Experiments were also performed to determine the reactivity of CFHR4 antibodies with human CFHR3 protein (Figure 9). Binding experiments were performed in a Carterra LSA with a running buffer of PBS pH 7.40, 1% BSA, 0.05% Tween 20. Antibodies were covalently bound and printed onto HC30M chips. The chips were activated with 33 mM s-NHS and 133 mM EDC in 100 mM MES pH 5.5 for 7 minutes. 10 mg / ml of antibody in acetate buffer pH 4.5 was used for printing for 10 minutes. The printed chips were then quenched with 1 M ethanolamine pH 8.5 for 7 minutes. In the reactivity assay, 550 nM ATX-P-58, human CFHL3-His was tested with 5 minutes of association followed by 15 minutes of dissociation. Results were processed and analyzed with Carterra LSA Kinetics Software. Data was referenced to an intervening reference spot, double referenced to buffer cycles, and then responses (nm) after association were reported. Isotype controls were used to determine the cutoff response for positive binding.

[0178] Experiments were also performed to determine the reactivity of the CFHR4 antibody with human CFHL-1 (Y402H) protein (Figure 10). Binding experiments were performed in a Carterra LSA with a running buffer of PBS pH 7.40, 1% BSA, 0.05% Tween 20. Antibodies were covalently bound and printed onto HC30M chips. The chips were activated with 33 mM s-NHS and 133 mM EDC in 100 mM MES pH 5.5 for 7 minutes. 10 mg / ml of antibody in acetate buffer pH 4.5 was used for printing for 10 minutes. The printed chips were then quenched with 1 M ethanolamine pH 8.5 for 7 minutes. In the reactivity assay, 380 nM ATX-P-421, human CFHL1-Y402H-His was tested with 5 minutes of association followed by 15 minutes of dissociation. Results were processed and analyzed with Carterra LSA Kinetics Software. Data was referenced to an intervening reference spot, double referenced to buffer cycles, and then responses (nm) after association were reported. Isotype controls were used to determine the cutoff response for positive binding.

[0179] Example 8 C3 Convertase Assembly Assay. A screening assay was developed to assess the ability of the disclosed CFHR4 antibodies to affect C3 convertase formation (FIG. 11A). C3bBb is the C3 convertase (EC 3.4.21.43) of the alternative pathway of complement activation. The C3 enzyme is involved in amplifying pathway activation and depositing C3b and the membrane attack complex on target cells. It is regulated by serum proteins factor H, factor I (EC 3.4.21.45), and properdin. Formation of C3 convertase requires C3b, factor B, factor D (EC 3.4.21.46), and Mg2+. After formation of the reversible bimolecular complex C3bB(Mg), factor D cleaves factor B, releasing the activation fragment Ba and generating the active enzyme C3bBb. The C3 enzyme is a serine protease with a catalytic site present in the Bb subunit. Furthermore, CFHR4 has been reported to directly promote C3 convertase formation by binding to C3b, has a higher ability to assemble C3bBb on CFHR4, and the C3bBb-CFHR4 complex is more resistant to CFH cleavage compared with C3bBb.

[0180] Therefore, the assay is designed to test the ability of CHFR4 antibody to disrupt the formation of C3 convertase complex by measuring the amount of factor B present using anti-FB monoclonal antibody conjugated to a reporter moiety.As illustrated in Figure 11A, in the presence of CFHR4 (in the absence of effective CFHR4 antibody), C3 convertase complex is formed, anti-FB antibody binds to FB, and a signal is detected (i.e., complement activation).However, in the presence of effective anti-CFHR4 antibody, C3 convertase complex is disrupted, antibody-bound FB is washed away, and the signal is reduced (i.e., no complement activation).

[0181] An example of an assay protocol is provided below.

[0182] Preparation of FHR-4 coated plates - (1) Coat plates with 5 μg / ml FHR-4 in PBS and incubate with moistened paper O / N in a plastic bag. (2) Wash 3x300 μl with Wieslab wash buffer. (3) Block wells with 150 μl PBS containing 3% BSA. (4) Incubate for a minimum of 1 hour at room temperature. (5) Aspirate wells and wash with 1x300 μl Superblock. (6) Dry plates at 37°C, 16% RH for at least 1 hour. (7) Store in plate bag and refrigerate.

[0183] Assay protocol - (1) Bring diluent, plate, substrate, and stop solution to room temperature. (2) Dilute anti-OVA antibody to desired concentration in AP diluent. (3) Add 50 μl of anti-OVA, AP diluent (blank), and diluent control to plate. (4) Incubate 1 hour at room temperature with lid on. (5) Wash with 3×300 μl Wieslab wash buffer. (6) Add 50 μl of 5 μg / ml C3b or AP diluent to plate. (7) Incubate plate at 37° C. for 30 minutes. (8) Wash with 3×300 μl Wieslab wash buffer. (9) Add 50 μl of protein mix (FB, FD, properdin) to plate. (10) Incubate plate at 37° C. for 30 minutes. (11) Wash with 3×300 μl Wieslab wash buffer. (12) 50 μl of anti-factor B antibody diluted 1:2000 in AP diluent was added to the entire plate. (13) Plate was incubated for 1 hour at room temperature. (14) Washed with 3×300 μl Wieslab wash buffer. (15) 50 μl of rabbit anti-goat IgG-HRP antibody diluted 1:5000 in Stabilzyme was added to the entire plate. (16) Plate was incubated for 1 hour at room temperature. (17) Washed with 3×300 μl Wieslab wash buffer. (18) 50 μl of substrate was added to the entire plate. (19) Plate was incubated for 20 minutes at room temperature. (20) 50 μl of stop solution was added to the entire plate. (21) Plate was read at 450-620 nm.

[0184] Dose-response curves were generated for the top eight inhibitory antibodies screened in the C3 convertase assembly assay, as shown in Figure 11B. Percent C3 convertase activity is plotted relative to the negative control (diluent only). Figure 11C includes the calculated IC50 values ​​for each antibody tested in Figure 11B. Figure 11D provides a representative summary of the data showing at least three functional classes of anti-CFHR4 antibodies.

[0185] array

[0186] The various amino acid and nucleic acid sequences referenced herein are provided below.

[0187] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10]

Table 3-11

Table 3-12

Table 3-13

Table 3-14

Table 3-15

Table 3-16

Table 3-17

Table 3-18

Table 3-19

Table 3-20

[0188]

Table 4-1

Table 4-2

Table 4-3

Table 4-4

Table 4-5

Table 4-6

Table 4-7

Table 4-8

Table 4-9

Table 4-10

Table 4-11

Table 4-12

Table 4-13

Table 4-14

Table 4-15

Table 4-16

Table 4-17

Table 4-18

Table 4-19

Table 4-20

Table 4-21

Table 4-22

Table 4-23

Table 4-24

Table 4-25

Table 4-26

Table 4-27

Table 4-28

Table 4-29

Table 4-30

Table 4-31

Table 4-32

Table 4-33

Table 4-34

Table 4-35

Table 4-36

[0189]

Table 5-1

Table 5-2

Table 5-3

Table 5-4

Table 5-5

Table 5-6

Table 5-7

Table 5-8

Table 5-9

Table 5-10

Table 5-11

Table 5-12

Table 5-13

Table 5-14

Table 5-15

Table 5-16

Table 5-17

Table 5-18

Table 5-19

Table 5-20

Table 5-21

Table 5-22

Table 5-23

Table 5-24

Table 5-25

Table 5-26

Table 5-27

Table 5-28

Table 5-29

Table 5-30

Table 5-31

Table 5-32

Table 5-33

Table 5-34

Table 5-35

Table 5-36

Table 5-37

Table 5-38

Table 5-39

Table 5-40

Table 5-41

Table 5-42

Table 5-43

Table 5-44

Table 5-45

Table 5-46

Table 5-47

Table 5-48

Table 5-49

Table 5-50

Table 5-51

Table 5-52

Table 5-53

Table 5-54

Table 5-55

Table 5-56

Table 5-57

Table 5-58

Table 5-59

Table 5-60

Table 5-61

Table 5-62

Table 5-63

Table 5-64

Table 5-65

Table 5-66

Table 5-67

Table 5-68

Table 5-69

Table 5-70

Table 5-71

Table 5-72

Table 5-73

Table 5-74

Table 5-75

Table 5-76

Table 5-77

Table 5-78

Table 5-79

Table 5-80

Table 5-81

Table 5-82

Table 5-83

Table 5-84

Table 5-85

Table 5-86

[0190] Human complement factor H related protein 4b (CFHR4b) (see, e.g., UniProt Accession No. Q92496): MLLLINVILTLWVSCANGQEVKPCDFPEIQHGGLYYKSLRRLYFPAAAGQSYSYYCDQNFVTPSGSYWDYIHCTQDGWSPTVPCLRTCSKSDIEIENGFISESSSIYILNKEIQYKCKPGYATADGNSSGSITCLQNGWSAQPICIKFCDMPVFENSRAKSNGMRFKLHDTLDYECYDGYEISYGNTTGSIVCGEDGWSHFPTCYNSSEKCGPPPPISNGDTTSFLLKVYVPQSRVEYQCQSYYELQGSNYVTCSNGEWSEPPRCIHPCIITEENMNKNNIQLKGKSDIKYYAKTGDTIEFMCKLGYNANTSVLSFQAVCREGIVEYPRCE (SEQ ID NO: 580).

[0191] Human CFHL1-Y402H-His(ATX-P-421): MYRMQLLSCIALSLALVTNSEDCNELPPRRNTEILTGSWSDQTYPEGTQAIYKCRPGYRSLGNVIMVCRKGEWVALNPLRKCQKRPCGHPGDTPFGTFTLT GGNVFEYGVKAVYTCNEGYQLLGEINYRECDTDGWTNDIPICEVVKCLPVTAPENGKIVSSAMEPDREYHFGQAVRFVCNSGYKIEGDEEMHCSDDGFWSKEKPKCVEISCKSPDVINGSPISQKIIYKE NERFQYKCNMGYEYSERGDAVCTESGWRPLPSCEEKSCDNPYIPNGDYSPLRIKHRTGDEITYQCRNGFYPATRGNTAKCTSTGWIPAPRCTLKPCDYPDIKHGGLYHENMRRPYFPVAVGKYYSYYCDEHFETPSGSYWDHIHCTQDGWSPAVPCLRKCYFPYLENGYNQNHGRKFVQGKSIDVACHPGYALPKAQTTVTCMENGWSPTPRCIRVSFTLGGGGSGLNDIFEAQKIEWHEGGGGSHHHHHH (sequence number 581).

[0192] [Table 6-1] [Table 6-2]

[0193] Various embodiments of the present disclosure are described herein. Variations of those embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors expect that those skilled in the art will utilize such variations as necessary, and the inventors intend that the various embodiments of the present disclosure may be practiced otherwise than as specifically described herein. Accordingly, the embodiments of the present disclosure include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed in the various embodiments of the present disclosure, unless otherwise indicated herein or clearly contradicted by context.

Claims

1. An antibody, or an antigen-binding fragment thereof, that specifically binds to human complement H factor-related (CFHR) 4, wherein the human CFHR 4 is optionally a CFHR 4 b variant (CFHR 4b), and optionally a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:

580.

2. The antibody or fragment according to claim 1, exhibiting one or more of the following functional characteristics: a. To reduce complement activation as optionally measured by an in vitro C3 convertase assembly assay, and / or to attenuate complement component 3 (C3) convertase activity, and / or compared to the absence of the antibody or fragment. b. Cross-reacting with at least one of the following: cynomolgus monkey CFHR4b protein (cCFHR4b), human CFHR4a variant (CFHR4a), human complement H factor-related protein 3 (CFHR3), and / or complement H factor-related protein 1 (CFHL1), and / or c. Not cross-reacting with at least one of the following: cynomolgus monkey CFHR4b protein (cCFHR4b), human CFHR4a variant (CFHR4a), human complement H factor-related protein 3 (CFHR3), and / or complement H factor-related protein 1 (CFHL1), and / or d. Human CFHR4 with K100 nM or less Ｄ To join together, and / or e. The sequence binds to the same human CFHR4 epitope as an antibody containing the VH and VL sequences of any one of the exemplary antibodies presented in Table 6, and / or f. The antibody competes for binding to human CFHR4 with an antibody containing the VH and VL sequences of any one of the exemplary antibodies whose sequences are presented in Table 6.

3. An antibody of the fragment according to claim 1 or 2, which is monoclonal and optionally recombinant.

4. An antibody of the fragment according to claim 1, which is human, humanized, or chimeric.

5. The antibody or fragment according to claim 1, wherein the antibody is a full-length antibody, a single-chain antibody, a single-chain variable fragment (scFv), a variable fragment (Fv), an antigen-binding region (Fab), Fab-C, Fab'-SH, (Fab')2, a single-domain antibody (sdAb), a VHH antibody, a nanobody, a single-domain antibody derived from a camelid, a single-domain antibody fragment (VNAR) derived from a shark IgNAR, a diabody, a triabody, an antikalin, or an aptamer, wherein the antibody is a full-length antibody containing an Fc region, for example, a human IgG1, IgG2, IgG3, or IgG4 region.

6. The antibody or fragment according to claim 1, which is conjugated to at least one additional portion optionally selected from the following: a. An antigen-binding portion, such as an antibody or its antigen-binding fragment, that can specifically bind to a target other than human CFHR4, wherein the target is expressed in the human eye, the antigen-binding portion, b. The area to be treated or the cytotoxic area, c. Detection section, d. Purification part, e. The half-life extension portion is optionally a polypeptide having at least 20 amino acids in length and comprising any combination of G, A, ST, E, and P residues, which is conjugated to the C or N terminus of the antibody.

7. The antibody or fragment according to Claim 1, a. One, two, or all three HCDRs of any one of the exemplary antibodies whose sequences are presented in Table 6, and optionally, one, two, or all three of the corresponding LCDRs of the exemplary antibodies, and / or b. A VH sequence having at least 90% identity with the VH sequence of any one of the exemplary antibodies whose sequences are presented in Table 6, and optionally a VL sequence having at least 90% identity with the corresponding VL sequence of the exemplary antibody, preferably the VH sequence which is not subject to change in the HCDR or LCDR, and optionally the VL sequence, and / or c. All six CDRs of any one of the exemplary antibodies whose sequences are presented in Table 6, and / or d. The VH and VL sequences of any one of the exemplary antibodies whose sequences are presented in Table 6, and / or e. The antibody, or a fragment thereof, is a polypeptide comprising the full-length heavy chain (VH+ constant) sequence of any one of the exemplary antibodies whose sequences are presented in Table 6, and optionally the corresponding full-length light chain (VL+ constant) sequence of the exemplary antibody.

8. A polynucleotide encoding an antibody or fragment according to Claim 1, wherein the polynucleotide optionally comprises or comprises a nucleic acid sequence whose sequence is at least 70, 80, 90, or 100% identical to any one of the exemplary antibodies presented in Table 6.

9. An expression vector comprising a polynucleotide according to claim 8, which is optionally an adeno-associated virus (AAV) vector, a lentivirus (LV) vector, a herpes simplex virus (HSV) vector, or a retroviral vector.

10. The antibody or fragment, polynucleotide, or vector according to Claim 1, and optionally, a. At least one pharmaceutically acceptable carrier, diluent, or preservative, and / or b. A pharmaceutical composition comprising at least one additional active ingredient.

11. The pharmaceutical composition according to claim 10, which is suitable for transocular administration to a subject, optionally by delivery using conjunctival implants, contact lenses, gels, nanoparticles, mucosal adhesion polymers, ointments, solutions, suspensions, eye drops, and / or implants, preferably by injection into vitreous solution.

12. An antibody or fragment according to claim 1, a polynucleotide according to claim 8, a vector according to claim 9, or a composition according to claim 10 or 11, for use as a pharmaceutical, or optionally for use in a method for treating a disease of the eye in question.

13. An antibody, fragment, polynucleotide, vector, or composition for use according to claim 12, characterized in that the disease is characterized by increased activation of the complement system, particularly alternative pathways, and particularly increased activation in the eye of the subject, for example, in dlusen or retinal pigment epithelial (RPE) cells of the subject.

14. The method comprises, preferably, transocular administration of the antibody by injection into a vitreous solution, wherein the administration preferably alleviates at least one symptom in the subject selected from visual distortion, reduced central vision, blurred vision, and / or difficulty adapting to low light, the antibody, fragment, polynucleotide, vector, or composition for use according to claim 12.

15. The disease is age-related macular degeneration (AMD), and optionally, the AMD is dry AMD and may be in an early, mid, or progressive stage (the latter also known as geographic atrophy or GA), an antibody, fragment, polynucleotide, vector, or composition for use according to claim 12.

16. An antibody that specifically binds to human complement factor H-related 4 (CFHR4), or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) including complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and a light chain variable region (VL) including complementarity-determining regions (CDRs) LCDR1, LCDR2, and LCDR3, (a) HCDR1 comprises an amino acid sequence selected from SEQ ID NOs: 1-5, 12-20, 53-60, or 88-94; (b) HCDR2 comprises an amino acid sequence selected from SEQ ID NOs: 8-14, 32-42, 65-76, or 102-115; (c) HCDR3 comprises an amino acid sequence selected from SEQ ID NOs: 15-20, 43-52, 77-87, or 116-128; (d) LCDR1 comprises an amino acid sequence selected from SEQ ID NOs: 129-151, 197-199, 206-215, or 236-243; (e) LCDR2 comprises an amino acid sequence selected from SEQ ID NOs: 152-174, 200-202, 216-225, or 245-251; and (f) The antibody or its antigen-binding fragment, wherein LCDR3 comprises an amino acid sequence selected from SEQ ID NOs: 175-196, 204-205, 227-235, or 253-259.

17. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment comprises HCDR1 of SEQ ID NO: 2, HCDR2 of SEQ ID NO: 9, HCDR3 of SEQ ID NO: 15, LCDR1 of SEQ ID NO: 129, LCDR2 of SEQ ID NO: 152, and LCDR3 of SEQ ID NO:

175.

18. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment comprises HCDR1 of SEQ ID NO: 5, HCDR2 of SEQ ID NO: 12, HCDR3 of SEQ ID NO: 18, LCDR1 of SEQ ID NO: 206, LCDR2 of SEQ ID NO: 216, and LCDR3 of SEQ ID NO:

226.

19. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment that binds to CFHR4 inhibits or reduces CFHR4-mediated complement activation, or interferes with the binding of CFHR4 to complement component 3 (C3) or complement factor H (CFH).

20. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment binds to CFHR4 with a dissociation constant (KD) of less than 1 × 10⁻⁸ M as measured by surface plasmon resonance.

21. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment is a full-length human IgG1, IgG2, IgG3, or IgG4 antibody.

22. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment comprises a single-chain antibody, a single-chain variable fragment (scFv), a variable fragment (Fv), an antigen-binding region (Fab), Fab-C, Fab'-SH, (Fab')2, a single-domain antibody (sdAb), a VHH antibody, a nanobody, a single-domain antibody derived from a camelid, a single-domain antibody fragment (VNAR) derived from a shark IgNAR, a diabody, a triabody, an antikalin, or an aptamer.

23. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment is of human origin, humanized, or chimeric.

24. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment competes for binding to CFHR4 with an antibody comprising the VH sequence of SEQ ID NO: 260 and the VL sequence of SEQ ID NO:

272.

25. The antibody or antigen-binding fragment according to claim 16, wherein the antibody or antigen-binding fragment is conjugated to one or more portions independently selected from the following: a. Antigen-binding portion of an antibody or its antigen-binding fragment that can specifically bind to a target other than human CFHR4. b. The area to be treated or the cytotoxic area, c. Detection section, d. Purified portion, and / or e. The half-life extension portion is optionally a polypeptide having a length of at least 20 amino acids and containing any combination of G, A, ST, E, and P residues, which is conjugated to the C or N terminus of the antibody.

26. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to claim 16, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

27. ​​The pharmaceutical composition according to claim 26, wherein the pharmaceutical composition is suitable for transocular administration.

28. The pharmaceutical composition according to claim 27, wherein the transocular administration includes delivery using a conjunctival implant, a contact lens, a gel, nanoparticles, a mucosal adhesive polymer, an ointment, a solution, a suspension, eye drops, or an implant.

29. The pharmaceutical composition according to claim 26 for the treatment of age-related macular degeneration (AMD).

30. The pharmaceutical composition according to claim 29, which is administered transocularly to treat at least one symptom of AMD.

31. The pharmaceutical composition according to claim 29, wherein the AMD comprises a wet-type AMD or a dry-type AMD.

32. The pharmaceutical composition according to claim 30, wherein the at least one AMD symptom includes visual distortion, reduced central vision, blurred vision, and difficulty adapting to low light.

33. The pharmaceutical composition according to claim 29, administered to reduce complement activation in the eye of a subject.

34. The pharmaceutical composition according to claim 29, administered in a dose ranging from about 0.0001 mg / dose to about 100 mg / dose.

35. The pharmaceutical composition according to claim 29, administered in a dose ranging from about 0.0001 mg / ml to about 100 mg / ml.