WO2018237338A1

WO2018237338A1 - Anti-alpha-synuclein antibodies and methods of use thereof

Info

Publication number: WO2018237338A1
Application number: PCT/US2018/039116
Authority: WO
Inventors: Rui Hao; Suresh PODA; Rishi Rakhit; Rinkan SHUKLA; Adam P. Silverman
Original assignee: Denali Therapeutics Inc.
Priority date: 2017-06-23
Filing date: 2018-06-22
Publication date: 2018-12-27

Abstract

In one aspect, antibodies that specifically bind to a human alpha-synuclein protein are provided. In some embodiments, an anti-alpha-synuclein antibody specifically binds to monomeric human alpha-synuclein, oligomeric human alpha-synuclein, human alpha-synuclein fibrils, and/or soluble human alpha-synuclein. In some embodiments, the anti-alpha-synuclein antibody is a pan-alpha-synuclein antibody.

Description

ANTI-ALPHA-SYNUCLEIN ANTIBODIES AND METHODS OF USE

THEREOF

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] The present application claims priority to U.S. Provisional Application No. 62/524,462, filed on June 23, 2017, and U.S. Provisional Application No. 62/583,430, filed on November 8, 2017, the disclosures of which are incorporated herein by reference in their entirety for all purposes.

BACKGROUND OF THE INVENTION

[0002] Alpha-synuclein protein has been identified as a significant component of Lewy bodies and Lewy neurites, which are defining neuropathological characteristics of Parkinson's disease and other neurodegenerative diseases that are classified as synucleinopathies. In addition, the filamentous inclusions that are found in multiple system atrophy are made of alpha-synuclein. Furthermore, alpha-synuclein has been implicated in the pathogenesis of a large number of neurodegenerative diseases that are classified as tauopathies, such as Alzheimer's disease, following the discovery that there are interactions between alpha- synuclein and tau proteins.

[0003] It is believed that the aggregation of alpha-synuclein oligomers and the formation of alpha-synuclein fibrils, followed by the spreading of alpha-synuclein species, results in the formation of Lewy bodies, which in turn contribute to neural cell toxicity and death. Accordingly, there is a need for therapeutic agents that target and neutralize alpha-synuclein protein, thus preventing the aggregation and oligomerization of alpha-synuclein protein as well as preventing the spreading of alpha-synuclein oligomers and fibrils.

BRIEF SUMMARY OF THE INVENTION

[0004] In one aspect, antibodies (or antigen-binding portions thereof) that specifically bind to a human alpha-synuclein protein are provided. In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 106-125 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 106-120 or residues 111-125 of SEQ ID NO: l . [0005] In some embodiments, the antibody comprises one or more complementarity determining regions (CDRs) selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:205 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:205; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:231 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:231.

[0006] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

[0007] In some embodiments, the antibody comprises: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:62; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO : 121.

[0008] In some embodiments, the antibody comprises: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:62; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:267.

[0009] In some embodiments, the antibody comprises: (a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205; (b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

[0010] In some embodiments, the antibody comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:62, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 121, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

[0011] In some embodiments, the antibody comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:62, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:267, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

[0012] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:2-3 and 266. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 146-148. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:2-3 and 266, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 146-148.

[0013] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:2 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 147. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:266 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 147.

[0014] In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 71-105 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 71-85, residues 86-100, or residues 91-105 of SEQ ID NO: 1.

[0015] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:83-115, 245, and 246 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:83- 115, 245, and 246; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249;

(d) a light chain CDRl having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:206-220 and 251 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:206-220 and 251; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:224-228 and 253 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:224-228 and 253; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:232-237 and 255 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:232-237 and 255.

[0016] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:83-115, 245, and 246; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249; (d) a light chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:206-220 and 251 ;

(e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:224- 228 and 253; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232-237 and 255.

[0017] In some embodiments, the antibody comprises: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:65; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:88; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 126.

[0018] In some embodiments, the antibody comprises: (a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:218; (b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228; and (c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0019] In some embodiments, the antibody comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:88, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 126, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0020] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:4-55. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 149-197. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:4-55, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 149-197. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 12 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 157.

[0021] In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 66-85 of SEQ ID NO: 1. In some embodiments, the antibody or antigen- binding portion thereof recognizes an epitope within residues 66-80 or residues 71-85 of SEQ ID NO: l . [0022] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:79 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 79; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 116 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 116; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 142 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 142; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:221; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:229 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:229; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:238 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:238.

[0023] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:79; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:221; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

[0024] In some embodiments, the antibody comprises: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:79; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142.

[0025] In some embodiments, the antibody comprises: (a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:221; (b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229; and (c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

[0026] In some embodiments, the antibody comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:79, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:221, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

[0027] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:56. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 198. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:56, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 198.

[0028] In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 81-115 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 81-95, residues 86-100, residues 91-105, or residues 101-115 of SEQ ID NO: l .

[0029] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 117, 118, and 247 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 117, 118, and 247; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:237 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:237.

[0030] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: l 17, 118, and 247; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0031] In some embodiments, the antibody comprises: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:65; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 118; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 144.

[0032] In some embodiments, the antibody comprises: (a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:219; (b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:230; and (c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0033] In some embodiments, the antibody comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 118, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 144, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:219, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:230, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0034] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:57-58. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 199-201. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 57-58, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 199-201. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:58 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:201.

[0035] In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 36-50 and residues 86-115 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 36-50, residues 86-100, residues 96-110, or residues 101-115 of SEQ ID NCv l .

[0036] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:63 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 63; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 119, 120, and 248 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 119, 120, and 248; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 145 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 145; (d) a light chain CDRl having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:220; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:228 or 316 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:228 or 316; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:239 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:239.

[0037] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:63; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 119, 120, and 248; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:220; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228 or 316; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239. [0038] In some embodiments, the antibody comprises: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 63; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 119; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145.

[0039] In some embodiments, the antibody comprises: (a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:220; (b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228; and (c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

[0040] In some embodiments, the antibody comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:63, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 119, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

[0041] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:59-61. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:202-204. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:59-61, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:202-204. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:60 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:203.

[0042] In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 86-105 or residues 121-140 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 86-100, residues 91-105, residues 121-135, or residues 126-140 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 86-105 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 121-140 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 126-140 of SEQ ID NO: l . In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 86-105 and 121-140 of SEQ ID NO: l .

[0043] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:279, 281-284 and 311-312 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:279, 281- 284, and 311-312; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315.

[0044] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 311-312; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315. [0045] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:269-272 and 274-275. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:292-295 and 298-300. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:269-272 and 274-275, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:292-295 and 298-300.

[0046] In some embodiments, the antibody or antigen-binding portion thereof recognizes an epitope within residues 61-140 of SEQ ID NO: l .

[0047] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:277 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:280 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:280; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:289 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:289; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:304 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 304; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:308 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:308.

[0048] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:280; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:289; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:304; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 308.

[0049] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:273. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:296- 297. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:273, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:296-297.

[0050] In some embodiments, the antibody comprises a first Fc polypeptide and optionally a second Fc polypeptide. In some embodiments, the antibody comprises the first Fc polypeptide and the second Fc polypeptide. In some embodiments, the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide.

[0051] In some embodiments, the antibody comprises: (a) a first antigen-binding portion comprising a first variable region that specifically binds to the human alpha-synuclein protein, wherein the first antigen-binding portion comprises (i) a first heavy chain comprising a first Fc polypeptide and (ii) a first light chain; and (b) a second antigen-binding portion comprising a second variable region that specifically binds to the human alpha-synuclein protein, wherein the second antigen-binding portion comprises (i) a second heavy chain comprising a second Fc polypeptide and (ii) a second light chain; wherein the first Fc polypeptide and the second Fc polypeptide form an Fc dimer. In some embodiments, the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide.

[0052] In some embodiments, the first variable region and the second variable region recognize the same epitope in the human alpha-synuclein protein. In some embodiments, the first variable region and the second variable region recognize different epitopes in the human alpha-synuclein protein. In some embodiments, the first Fc polypeptide and the second Fc polypeptide each contain modifications that promote heterodimerization. In some embodiments, one of the Fc polypeptides has a T366W substitution and the other Fc polypeptide has T366S, L368A, and Y407V substitutions, according to EU numbering.

[0053] In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises a native FcRn binding site. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises a modification that alters FcRn binding. In some embodiments, the first Fc polypeptide and the second Fc polypeptide do not have effector function. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide includes a modification that reduces effector function. In some embodiments, the modification that reduces effector function comprises the substitutions of Ala at position 234 and Ala at position 235, according to EU numbering.

[0054] In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises amino acid changes relative to the native Fc sequence that extend serum half-life. In some embodiments, the amino acid changes comprise substitutions of Tyr at position 252, Thr at position 254, and Glu at position 256, according to EU numbering. Alternatively, in other embodiments, the amino acid changes comprise substitutions of Leu at position 428 and Ser at position 434, according to EU numbering. Alternatively, in further embodiments, the amino acid changes comprise a substitution of Ser or Ala at position 434, according to EU numbering.

[0055] In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide specifically binds to a transferrin receptor. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises at least two substitutions at positions selected from the group consisting of 384, 386, 387, 388, 389, 390, 413, 416, and 421, according to EU numbering. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises substitutions at least three, four, five, six, seven, eight, or nine of the positions. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide further comprises one, two, three, or four substitutions at positions comprising 380, 391, 392, and 415, according to EU numbering. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide further comprises one, two, or three substitutions at positions comprising 414, 424, and 426, according to EU numbering. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises Tip at position 388. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises an aromatic amino acid at position 421. In some embodiments, the aromatic amino acid at position 421 is Trp or Phe.

[0056] In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises at least one position selected from the following: position 380 is Trp, Leu, or Glu; position 384 is Tyr or Phe; position 386 is Thr; position 387 is Glu; position 388 is Trp; position 389 is Ser, Ala, Val, or Asn; position 390 is Ser or Asn; position 413 is Thr or Ser; position 415 is Glu or Ser; position 416 is Glu; and position 421 is Phe. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 positions selected from the following: position 380 is Trp, Leu, or Glu; position 384 is Tyr or Phe; position 386 is Thr; position 387 is Glu; position 388 is Trp; position 389 is Ser, Ala, Val, or Asn; position 390 is Ser or Asn; position 413 is Thr or Ser; position 415 is Glu or Ser; position 416 is Glu; and position 421 is Phe. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises 11 positions as follows: position 380 is Trp, Leu, or Glu; position 384 is Tyr or Phe; position 386 is Thr; position 387 is Glu; position 388 is Trp; position 389 is Ser, Ala, Val, or Asn; position 390 is Ser or Asn; position 413 is Thr or Ser; position 415 is Glu or Ser; position 416 is Glu; and position 421 is Phe.

[0057] In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide has a CH3 domain with at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 111-217 of any one of SEQ ID NOs:336-372 and 467-538. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:467-538. In some embodiments, the residues at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 of the positions corresponding to positions EU index 380, 384, 386, 387, 388, 389, 390, 391, 392, 413, 414, 415, 416, 421, 424 and 426 of any one of SEQ ID NOs:336-372 and 467-538 are not deleted or substituted.

[0058] In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide binds to the apical domain of the transferrin receptor. In some embodiments, the binding of the antibody or antigen-binding portion thereof to the transferrin receptor does not substantially inhibit binding of transferrin to the transferrin receptor. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide has an amino acid sequence identity of at least 75%), or at least 80%>, 90%, 92%, or 95%, as compared to the corresponding wild-type Fc polypeptide. In some embodiments, the corresponding wild-type Fc polypeptide is a human IgGl, IgG2, IgG3, or IgG4 Fc polypeptide.

[0059] In some embodiments, uptake of the antibody or antigen-binding portion thereof into the brain is greater than the uptake of the antibody or antigen-binding portion thereof without the modifications in the first Fc polypeptide and/or the second Fc polypeptide that result in transferrin receptor binding. In some embodiments, uptake of the antibody or antigen-binding portion thereof into the brain is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100-fold greater as compared to the uptake of the antibody or antigen-binding portion thereof without the modifications in the first Fc polypeptide and/or the second Fc polypeptide that result in transferrin receptor binding.

[0060] In other embodiments, one of the Fc polypeptides of the antibody is not modified to bind to a blood-brain barrier receptor and the other Fc polypeptide of the antibody is modified to specifically bind to a transferrin receptor.

[0061] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 20 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 20 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 100 pM to about 20 nM.

[0062] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 10 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 10 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 100 pM to about 10 nM.

[0063] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 5 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 5 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 100 pM to about 5 nM.

[0064] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 1 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 1 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 100 pM to about 1 nM.

[0065] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to monomeric and/or oligomeric human alpha-synuclein protein. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to monomeric and oligomeric human alpha-synuclein protein. In some embodiments, the antibody or antigen-binding portion thereof has a higher binding affinity for oligomeric human alpha-synuclein protein than for monomeric human alpha-synuclein protein. In some embodiments, the antibody or antigen- binding portion thereof has a higher binding affinity for monomeric human alpha-synuclein protein than for oligomeric human alpha-synuclein protein. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to soluble human alpha- synuclein protein and/or human alpha-synuclein protein fibrils. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to soluble human alpha- synuclein protein and human alpha-synuclein protein fibrils.

[0066] In some embodiments, the antibody is a pan-alpha-synuclein antibody or antigen- binding portion thereof. In some embodiments, the pan-alpha-synuclein antibody or antigen- binding portion thereof specifically binds to monomeric human alpha-synuclein protein, oligomeric human alpha-synuclein protein, soluble human alpha-synuclein protein, and human alpha-synuclein protein fibrils.

[0067] In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a fully human antibody. In some embodiments, the antigen-binding portion is a Fab, a F(ab')₂, a scFv, or a bivalent scFv.

[0068] In some embodiments, the antibody is a multispecific antibody. In some embodiments, the multispecific antibody is a bispecific antibody. In some embodiments, the bispecific antibody recognizes two different alpha-synuclein epitopes.

[0069] In another aspect, antigen-binding fragments that specifically bind to a human alpha- synuclein protein are provided. In some embodiments, the antigen-binding fragment further comprises an Fc polypeptide. In some embodiments, the Fc polypeptide is a modified Fc polypeptide. In some embodiments, the Fc polypeptide contains one or more of the modifications described herein, e.g., to promote heterodimerization, reduce effector function, extend serum half-life, and/or bind to a transferrin receptor. As a non-limiting example, the antigen-binding fragment may include a Fab fragment that further comprises an Fc polypeptide, e.g., a Fab-Fc fusion. In other embodiments, the antigen-binding fragment further comprises a first Fc polypeptide and a second Fc polypeptide. In some embodiments, the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide. In some embodiments, the first Fc polypeptide and/or the second Fc polypeptide contains one or more of the modifications described herein, e.g., to promote heterodimerization, reduce effector function, extend serum half-life, and/or bind to a transferrin receptor. As a non-limiting example, the antigen-binding fragment may include a F(ab')₂ fragment that further comprises a first Fc polypeptide and a second Fc polypeptide, e.g., F(ab')₂-Fc fusion.

[0070] In another aspect, pharmaceutical compositions are provided. In some embodiments, the pharmaceutical composition comprises an antibody that specifically binds to a human alpha-synuclein protein as described herein and further comprises one or more pharmaceutically acceptable carriers.

[0071] In yet another aspect, isolated polynucleotides are provided. In some embodiments, the isolated polynucleotide comprises a nucleotide sequence encoding an isolated antibody that specifically binds to a human alpha-synuclein protein as described herein. In another aspect, vectors and host cells comprising such an isolated polynucleotide are provided.

[0072] In yet another aspect, antibodies are provided that compete for specific binding to a human alpha-synuclein protein with an antibody as described herein.

[0073] In yet another aspect, hybridoma cell lines that secrete antibodies of the present invention are provided. In some embodiments, the hybridoma cell line is selected from the group consisting of 3D5, 4F5, 46D2, 28B6, 3E12, 1F4, 1G4, and 16D4. In another aspect, antibodies that are secreted by hybridoma cell lines of the present invention are provided.

[0074] In still another aspect, methods of neutralizing human alpha-synuclein protein in a brain of a subject are provided. In some embodiments, the method comprises administering to the subject an antibody (or antigen-binding portion thereof) that specifically binds to a human alpha-synuclein protein as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein. In some embodiments, the method of neutralizing human alpha-synuclein protein comprises preventing or reducing human alpha-synuclein protein oligomerization and/or aggregation in the brain of the subject. In some embodiments, the method of neutralizing human alpha-synuclein protein comprises preventing or reducing the spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject. [0075] In some embodiments, the isolated antibody, the pharmaceutical composition, or the bispecific antibody is administered prior to the broad spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject. In some embodiments, administering the isolated antibody, the pharmaceutical composition, or the bispecific antibody prior to the broad spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject increases the ability of the isolated antibody, the pharmaceutical composition, or the bispecific antibody to treat a neurodegenerative disease.

[0076] In some embodiments, the antibody, pharmaceutical composition, or bispecific antibody neutralizes monomelic human alpha-synuclein protein, oligomeric human alpha- synuclein protein, soluble human alpha-synuclein protein, human alpha-synuclein protein fibrils, or a combination thereof. In some embodiments, the antibody or bispecific antibody is a pan-alpha-synuclein antibody or a pan-alpha-synuclein bispecific antibody. In some embodiments, the subject has a neurodegenerative disease (e.g., a synucleinopathy).

[0077] In some embodiments, the isolated antibody, the pharmaceutical composition, or the bispecific antibody is administered during an early stage of the neurodegenerative disease. In some embodiments, the subject has not been treated for the neurodegenerative disease, has not been treated with a dopaminergic replacement therapy, or has not been treated with a dopamine agonist therapy.

[0078] In some embodiments, the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is necessary to neutralize the human alpha- synuclein protein in the brain of the subject is determined based on an amount of total human alpha-synuclein protein in a sample and/or an amount of human alpha-synuclein protein in a sample that is bound to the isolated antibody or bispecific antibody. In some embodiments, the sample is obtained from the subject. In some embodiments, the sample is selected from the group consisting of an interstitial brain fluid sample, a cerebrospinal fluid (CSF) sample, a blood sample, and a combination thereof.

[0079] In some embodiments, the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is necessary to neutralize the human alpha- synuclein protein in the brain of the subject is determined based on a level of neurodegenerative disease progression in the subject and/or an amount of human alpha-synuclein protein spreading in the brain of the subject. In some embodiments, an imaging method is used to determine the level of neurodegenerative disease progression in the subject and/or the amount of human alpha-synuclein protein spreading in the brain of the subject. In some embodiments, the imaging method is selected from the group consisting of positron emission tomography (PET) imaging, dopamine transporter (DaT) imaging, magnetic resonance imaging (MRI), and computed tomography (CT) imaging. In some embodiments, PET imaging comprises using a PET probe that comprises a diabody of an antibody as described herein.

[0080] In yet another aspect, methods of treating a neurodegenerative disease in a subj ect are provided. In some embodiments, the neurodegenerative disease is a synucleinopathy. In some embodiments, the method comprises administering to the subject an antibody (or antigen- binding portion thereof) that specifically binds to a human alpha-synuclein protein as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha- synuclein antibody as described herein.

[0081] In some embodiments, the neurodegenerative disease (e.g., synucleinopathy) is selected from the group consisting of Parkinson' s disease, dementia with Lewy bodies, multiple system atrophy, Huntington' s disease, Alzheimer' s disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down's syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden- Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle- predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick' s disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia.

[0082] In some embodiments, the subject is treated during an early stage of the neurodegenerative disease. In some embodiments, the subject has not been treated for the neurodegenerative disease, has not been treated with a dopaminergic replacement therapy, or has not been treated with a dopamine agonist therapy.

[0083] In some embodiments, the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is administered to the subject is determined based on an amount of total human alpha-synuclein protein in a sample and/or an amount of human alpha-synuclein protein in a sample that is bound to the isolated antibody or bispecific antibody. In some embodiments, the sample is obtained from the subject. In some embodiments, the sample is selected from the group consisting of an interstitial brain fluid sample, a cerebrospinal fluid (CSF) sample, a blood sample, and a combination thereof.

[0084] In some embodiments, the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is administered to the subject is based on a level of neurodegenerative disease progression in the subject and/or an amount of human alpha- synuclein protein spreading in the brain of the subject. In some embodiments, an imaging method is used to determine the level of neurodegenerative disease progression in the subject and/or the amount of human alpha-synuclein protein spreading in the brain of the subject. In some embodiments, the imaging method is selected from the group consisting of positron emission tomography (PET) imaging, dopamine transporter (DaT) imaging, magnetic resonance imaging (MRI), and computed tomography (CT) imaging. In some embodiments, PET imaging comprises using a PET probe that comprises a diabody of an antibody as described herein.

[0085] In another aspect, anti-alpha-synuclein antibodies for use in a method of neutralizing human alpha-synuclein protein in a brain of a subject are provided. In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein, is for use in a method of preventing or reducing human alpha-synuclein protein oligomerization and/or aggregation in the brain of the subject. In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein, is for use in a method of preventing or reducing the spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject.

[0086] In still another aspect, anti-alpha-synuclein antibodies for use in a method of treating a neurodegenerative disease are provided. In some embodiments, the neurodegenerative disease is a synucleinopathy. In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein, is for use in a method of treating a neurodegenerative disease (e.g., a synucleinopathy). In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein, is for use in a method of treating a neurodegenerative disease (e.g., a synucleinopathy) selected from the group consisting of Parkinson' s disease, dementia with Lewy bodies, multiple system atrophy, Huntington' s disease, Alzheimer's disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down' s syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden-Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle-predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick's disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia.

[0087] In yet another aspect, the use of an anti-alpha-synuclein antibody as described herein (or an antigen-binding portion thereof as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein) in the manufacture of a medicament for the neutralization of human alpha-synuclein protein in a brain of a subject is provided. In some embodiments, neutralizing comprises preventing or reducing human alpha-synuclein protein oligomerization and/or aggregation in the brain of the subject. In some embodiments, neutralizing comprises preventing or reducing the spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject.

[0088] In still another aspect, the use of an anti-alpha-synuclein antibody as described herein (or an antigen-binding portion thereof as described herein, or a pharmaceutical composition or bispecific antibody comprising an anti-alpha-synuclein antibody as described herein) in the manufacture of a medicament for the treatment of a neurodegenerative disease (e.g., a synucleinopathy) is provided. In some embodiments, the use of an anti-alpha-synuclein antibody as described herein is for the manufacture of a medicament for the treatment of a neurodegenerative disease (e.g., a synucleinopathy) selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Huntington's disease, Alzheimer's disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism- dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down's syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden-Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle-predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick's disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia.

BRIEF DESCRIPTION OF THE DRAWINGS

[0089] FIG. 1 shows the results of alpha-synuclein immunodepletion assays performed on cerebrospinal fluid obtained from Parkinson's disease patients.

[0090] FIG. 2 shows an assay design for the measurement of bound and free alpha-synuclein protein.

[0091] FIGS. 3 A and 3B show the results of bound and free alpha-synuclein assays for clones 2D02 and 2F10. FIG. 3 A shows the results of bound alpha-synuclein assays. FIG. 3B shows the results of free alpha-synuclein assays.

[0092] FIG. 4 shows a decrease in bound alpha-synuclein protein and a corresponding increase in free alpha-synuclein protein in wild-type mouse ISF.

DETAILED DESCRIPTION OF THE INVENTION I. INTRODUCTION

[0093] The present invention relates to the discovery of antibodies that have the ability to specifically bind to alpha-synuclein protein. In some embodiments, the anti-alpha-synuclein antibodies described herein specifically bind to monomeric human alpha-synuclein, oligomeric human alpha-synuclein, human alpha-synuclein fibrils, and/or soluble human alpha-synuclein. In some embodiments, the anti-alpha-synuclein antibodies described herein are pan-alpha- synuclein antibodies. In some embodiments, the anti-alpha-synuclein antibodies also exhibit cross-reactivity to mouse alpha-synuclein. In some embodiments, the anti-alpha-synuclein antibodies also exhibit cross-reactivity to cynomolgus monkey alpha-synuclein.

[0094] The anti-alpha-synuclein antibodies of the present invention are useful for, as non- limiting examples, neutralizing alpha-synuclein protein, inhibiting or preventing alpha- synuclein spreading, and inhibiting or preventing alpha-synuclein oligomerization and/or aggregation. Accordingly, antibodies of the present invention find utility for, among other things, preventing and treating various diseases such as degenerative neurological diseases.

II. DEFINITIONS

[0095] As used herein, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "an antibody" may include two or more such molecules, and the like.

[0096] As used herein, the terms "about" and "approximately," when used to modify an amount specified in a numeric value or range, indicate that the numeric value as well as reasonable deviations from the value known to the skilled person in the art, for example ± 20%, ± 10%, or ± 5%), are within the intended meaning of the recited value.

[0097] As used herein, the term "synuclein" or "synuclein protein" refers to a native (i.e., wild-type) alpha-synuclein protein of any vertebrate, including naturally occurring mutations and variants thereof, such as but not limited to human, non-human primates (e.g., cynomolgus monkey), rodents (e.g., mice), and other mammals. Unless stated otherwise, synuclein protein described herein includes protein prepared or obtained from tissue or fluid samples of any vertebrate, as well as from recombinant protein encoded by recombinant DNA and expressed from any expression cell or system. The term includes alpha-synuclein, beta-synuclein, and gamma-synuclein. The term also includes monomeric and oligomeric forms, soluble synuclein, and synuclein fibrils. As used herein, the term "alpha-synuclein" or "alpha- synuclein protein" refers to a "full-length" alpha-synuclein protein having a length of 140 amino acids (SEQ ID NO: 1). It will be recognized by a person of ordinary skill in the art that in humans, there are multiple isoforms of alpha-synuclein that result from alternative splicing of the gene encoding alpha-synuclein (SNCA). Examples of alpha-synuclein isoforms that result from alternative splicing include alpha-synuclein- 112 and alpha-synuclein-126. Non- limiting examples of alpha-synuclein amino acid sequences are set forth under UniProt reference identifier P37840. [0098] As used herein, the terms "an antibody that specifically binds to an alpha-synuclein protein" and "anti-alpha-synuclein antibody" interchangeably refer to an antibody that specifically binds to an alpha-synuclein protein (e.g., monomeric alpha-synuclein, oligomeric alpha-synuclein, soluble alpha-synuclein, alpha-synuclein fibrils, or an alpha-synuclein splice isoform). In some embodiments, an anti-alpha-synuclein antibody is an antibody that specifically binds to multiple forms of alpha-synuclein protein (e.g., monomeric alpha- synuclein, oligomeric alpha-synuclein, soluble alpha-synuclein, alpha-synuclein fibrils, and/or one or more alpha-synuclein splice isoforms). In some embodiments, an anti-alpha-synuclein antibody is a pan-alpha-synuclein antibody. As used herein, the term "pan-alpha-synuclein antibody" refers to an antibody that specifically binds to multiple forms of alpha-synuclein protein (e.g., monomeric alpha-synuclein, oligomeric alpha-synuclein, soluble alpha- synuclein, alpha-synuclein fibrils, or multiple alpha-synuclein splice isoforms). In an embodiment, a pan-alpha-synuclein antibody specifically binds to at least two of monomeric alpha-synuclein, oligomeric alpha-synuclein, soluble alpha-synuclein, alpha-synuclein fibrils, and multiple splice isoforms of alpha-synuclein. In an embodiment, a pan-alpha-synuclein antibody specifically binds to each of monomeric alpha-synuclein, oligomeric alpha-synuclein, soluble synuclein, alpha-synuclein fibrils, and multiple splice isoforms of alpha-synuclein. In an embodiment, the pan-alpha-synuclein antibody specifically binds to at least two of monomeric alpha-synuclein, oligomeric synuclein, soluble alpha-synuclein, alpha-synuclein fibrils, and multiple splice isoforms of alpha-synuclein, but may not bind to each form with the same affinity. In an embodiment, a pan-alpha-synuclein antibody specifically binds to each of monomeric alpha-synuclein, oligomeric alpha-synuclein, soluble alpha-synuclein, and multiple splice isoforms of alpha-synuclein.

[0099] As used herein, the term "antibody" refers to a protein with an immunoglobulin fold that specifically binds to an antigen via its variable regions. The term encompasses intact polyclonal antibodies, intact monoclonal antibodies, single chain antibodies, multispecific antibodies such as bispecific antibodies, monospecific antibodies, monovalent antibodies, chimeric antibodies, humanized antibodies, and human antibodies. The term "antibody," as used herein, also includes antibody fragments that retain antigen-binding specificity, including but not limited to Fab, F(ab')₂, Fv, scFv, and bivalent scFv. Antibodies can contain light chains that are classified as either kappa or lambda. Antibodies can contain heavy chains that are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. [0100] An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms "variable light chain" (VL) and "variable heavy chain" (VH) refer to these light and heavy chains, respectively.

[0101] The term "variable region" refers to a domain in an antibody heavy chain or light chain that is derived from a germline Variable (V) gene, Diversity (D) gene, or Joining (J) gene (and not derived from a Constant and C5) gene segment), and that gives an antibody its specificity for binding to an antigen. Typically, an antibody variable region comprises four conserved "framework" regions interspersed with three hypervariable "complementarity determining regions."

[0102] The term "complementarity determining region" or "CDR" refers to the three hypervariable regions in each chain that interrupt the four framework regions established by the light and heavy chain variable regions. The CDRs are primarily responsible for antibody binding to an epitope of an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3, numbered sequentially starting from the N-terminus, and are also typically identified by the chain in which the particular CDR is located. Thus, a VH CDR3 or CDR-H3 is located in the variable region of the heavy chain of the antibody in which it is found, whereas a VL CDR1 or CDR-L1 is the CDR1 from the variable region of the light chain of the antibody in which it is found.

[0103] The "framework regions" or "FRs" of different light or heavy chains are relatively conserved within a species. The framework region of an antibody, that is the combined framework regions of the constituent light and heavy chains, serves to position and align the CDRs in three-dimensional space. Framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences. For example, germline DNA sequences for human heavy and light chain variable region genes can be found in the "VBASE2" germline variable gene sequence database for human and mouse sequences.

[0104] The amino acid sequences of the CDRs and framework regions can be determined using various well known definitions in the art, e.g., Kabat, Chothia, international ImMunoGeneTics database (IMGT), AbM, and observed antigen contacts ("Contact"). In some embodiments, CDRs are determined according to the Contact definition. See, MacCallum et al., J. Mol. Biol., 262:732-745 (1996). In some embodiments, CDRs are determined by a combination of Kabat, Chothia, and Contact CDR definitions.

[0105] The terms "antigen-binding portion" and "antigen-binding fragment" are used interchangeably herein and refer to one or more fragments of a molecule, e.g., an antibody, that retains the ability to specifically bind to an antigen (e.g., an alpha-synuclein protein). Examples of antigen-binding fragments include, but are not limited to, a Fab fragment (a monovalent fragment consisting of the VL, VH, CL, and CHI domains), a F(ab')₂ fragment (a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region), a single chain Fv (scFv), a disulfide-linked Fv (dsFv), complementarity determining regions (CDRs), a VL (light chain variable region), a VH (heavy chain variable region), nanobodies, diabodies, each of which bind the antigen via a variable region, and other formats as described in Spiess et al., Mol. Immun. 67 (2015) 95-106, which is incorporated herein by reference, and any combination of these or any other functional portion of an immunoglobulin peptide capable of binding to a target antigen.

[0106] The term "epitope" refers to the area or region of an antigen to which a molecule, e.g. , the CDRs of an antibody, specifically binds and can include a few amino acids or portions of a few amino acids, e.g., 5 or 6, or more, e.g., 20 or more amino acids, or portions of those amino acids. In some cases, the epitope includes non-protein components, e.g., from a carbohydrate, nucleic acid, or lipid. In some cases, the epitope is a three-dimensional moiety. Thus, for example, where the target is a protein, the epitope can be comprised of consecutive amino acids {e.g., a linear epitope), or amino acids from different parts of the protein that are brought into proximity by protein folding {e.g., a discontinuous or conformational epitope). In some embodiments, an antibody specifically binds to two distinct regions of an antigen {e.g., an alpha-synuclein protein) that are not brought into proximity by protein folding, referred to herein as a "dual epitope." In some embodiments, the epitope is phosphorylated at one amino acid {e.g., at a serine or threonine residue).

[0107] As used herein, the phrase "recognizes an epitope," as used with reference to an anti- alpha-synuclein antibody, means that the antibody CDRs interact with or specifically bind to the antigen {i.e., the alpha-synuclein protein) at that epitope or a portion of the antigen containing that epitope. [0108] As used herein, the term "multispecific antibody" refers to an antibody that comprises two or more different antigen-binding portions, in which each antigen-binding portion comprises a different variable region that recognizes a different antigen, or a fragment or portion of the antibody that binds to the two or more different antigens. As used herein, the term "bispecific antibody" refers to an antibody that comprises two different antigen-binding portions, in which each antigen-binding portion comprises a different variable region that recognizes a different antigen, or a fragment or portion of the antibody that binds to the two different antigens. In some embodiments, a bispecific antibody comprises a first antigen- binding portion comprising a first variable region that recognizes an alpha-synuclein protein and a second antigen-binding portion comprising a second variable region that recognizes a non-alpha-synuclein antigen.

[0109] A "monoclonal antibody" refers to antibodies produced by a single clone of cells or a single cell line and consisting of or consisting essentially of antibody molecules that are identical in their primary amino acid sequence.

[0110] A "polyclonal antibody" refers to a pool of antibodies obtained from a heterogeneous population of antibodies in which different antibodies in the population bind to different epitopes of an antigen.

[0111] A "chimeric antibody" refers to an antibody molecule in which the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (i.e., variable region, CDR, or portion thereof) is linked to a constant region of a different or altered class, effector function and/or species, or in which the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity (e.g., CDR and framework regions from different species). In some embodiments, a chimeric antibody is a monoclonal antibody comprising a variable region from one source or species (e.g., mouse) and a constant region derived from a second source or species (e.g., human). Methods for producing chimeric antibodies are described in the art.

[0112] A "humanized antibody" is an antibody that retains the reactivity of a non-human antibody while being less immunogenic in humans. For example, a humanized antibody may contain a non-human derived variable region in which one or more residues are replaced with framework residues derived from a human germline. In some cases, some of the framework positions from the original species are retained, e.g., in order to retain the binding affinity and/or specificity of the non-human antibody once humanized. In some cases, some CDR residues may also be replaced with corresponding human residues.

[0113] A "human antibody" or a "fully human antibody" is an antibody having human heavy chain and light chain sequences, typically derived from human germline genes. In some embodiments, the antibody is produced by a human cell, by a non-human animal that utilizes human antibody repertoires (e.g., transgenic mice that are genetically engineered to express human antibody sequences), or by phage display platforms.

[0114] The term "specifically binds" refers to a molecule (e.g., an antibody (or an antigen- binding portion thereof) or a modified Fc polypeptide (or a target-binding portion thereof)) that binds to an epitope or target with greater affinity, greater avidity, and/or greater duration to that epitope or target in a sample than it binds to another epitope or non-target compound (e.g., a structurally different antigen). In some embodiments, an antibody (or an antigen-binding portion thereof) or a modified Fc polypeptide (or a target-binding portion thereof) that specifically binds to an epitope or target is an antibody (or an antigen-binding portion thereof) or a modified Fc polypeptide (or a target-binding portion thereof) that binds to the epitope or target with at least 5-fold greater affinity than other epitopes or non-target compounds, e.g., at least 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 25-fold, 50-fold, or 100-fold greater affinity. In some embodiments, an antibody that specifically binds to an alpha-synuclein protein binds to the alpha-synuclein protein with at least a 5-fold greater affinity than to a non- alpha-synuclein protein (e.g., at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, or at least 10-fold greater affinity). It will be recognized by one of skill that an antibody that specifically binds to a target (e.g., an alpha-synuclein protein) from one species may also specifically bind to orthologs of that target (e.g., an alpha-synuclein protein).

[0115] The term "binding affinity" is used herein to refer to the strength of a non-covalent interaction between two molecules, e.g., between an antibody (or an antigen-binding portion thereof) and an antigen, or between a modified Fc polypeptide (or a target-binding portion thereof) and a target. Thus, for example, the term may refer to 1 : 1 interactions between an antibody (or an antigen-binding portion thereof) and an antigen or between a modified Fc polypeptide (or a target-binding portion thereof) and a target, unless otherwise indicated or clear from context. Binding affinity may be quantified by measuring an equilibrium dissociation constant (KD), which refers to the dissociation rate constant (kd, time^"1) divided by the association rate constant (k_a, time^"1 M^"1). KD can be determined by measurement of the kinetics of complex formation and dissociation, e.g., using Surface Plasmon Resonance (SPR) methods, e.g., a Biacore™ system; kinetic exclusion assays such as KinExA®; and BioLayer interferometry (e.g., using the ForteBio® Octet platform). As used herein, "binding affinity" includes not only formal binding affinities, such as those reflecting 1 : 1 interactions between an antibody (or an antigen-binding portion thereof) and an antigen or between a modified Fc polypeptide (or a target-binding portion thereof) and a target, but also apparent affinities for which KD' S are calculated that may reflect avid binding.

[0116] As used herein, the term "neutralizing alpha-synuclein protein" means to inhibit or reduce the ability of an alpha-synuclein protein to bind to, or interact with, another moiety. As a non-limiting example, neutralization of an alpha-synuclein protein inhibits or reduces its ability to bind to or interact with other alpha-synuclein monomers, alpha-synuclein oligomers, alpha-synuclein fibrils, or soluble forms of alpha-synuclein. As another non-limiting example, neutralization of an alpha-synuclein protein inhibits or reduces its ability to bind to or interact with a non-alpha-synuclein protein, such as a tau protein or an enzyme (e.g., a kinase). In some embodiments, neutralizing alpha-synuclein protein comprises preventing the aggregation and/or oligomerization of alpha-synuclein protein (e.g., in a brain of a subject). In some embodiments, neutralizing alpha-synuclein protein comprises preventing the spreading of alpha-synuclein oligomers and/or fibrils (e.g., in a brain of a subject).

[0117] The terms "spreading of alpha-synuclein protein" and "alpha-synuclein protein spreading" refer to any increase in the amount of deposition of alpha-synuclein protein (e.g., in a brain of a subject) or any increase in the volume of an organ (e.g., a brain) in which alpha- synuclein protein is present. In some embodiments, alpha-synuclein protein spreading is continuous. As a non-limiting example, alpha-synuclein protein can be present (e.g., in a brain of a subject) at a particular site or locus, and the size (e.g., volume) of the site or locus can increase without an associated increase in the number of sites or loci at which alpha-synuclein is present. In some embodiments, alpha-synuclein protein spreading is discontinuous. As a non-limiting example, alpha-synuclein protein can be present at a particular site or locus, and as the spreading of alpha-synuclein protein progresses, the number of sites or loci at which alpha-synuclein is present increases, but the size of the original site or locus does not increase. In some embodiments, alpha-synuclein spreading is both continuous and discontinuous. As a non-limiting example, as alpha-synuclein protein progresses, the size of particular a particular site or locus at which alpha-synuclein is present can increase, and the number of sites or loci can increase. [0118] In some embodiments, alpha-synuclein protein spreading is associated with an increase in size (e.g., volume) of one more sites or loci at which alpha-synuclein is present, is associated with the spreading of alpha-synuclein beyond a particular structure or region of an organ (e.g., a brain of a subject), or a combination thereof. In some embodiments, alpha- synuclein protein spreading is associated with the onset, progression, or worsening of the signs or symptoms of a disease (e.g., a neurodegenerative disease such as a synucleinopathy). As non-limiting examples, symptoms of a neurodegenerative disease include memory deficits, cognitive deficits, motor deficits, sensory deficits, and speech deficits. Alpha-synuclein protein spreading can refer to the spreading of oligomeric alpha-synuclein, monomeric alpha- synuclein, soluble alpha-synuclein, and/or alpha-synuclein fibrils. In particular embodiments, alpha-synuclein spreading refers to the spreading of a toxic form of alpha-synuclein. In some instances, the toxic form of alpha-synuclein is oligomeric alpha-synuclein.

[0119] In some embodiments, the term "broad spreading" refers to the spreading of alpha- synuclein protein (e.g., a toxic form of alpha-synuclein protein) beyond a single region, area, or structure in an organ (e.g., a brain of a subject). In some embodiments, the term refers to the spreading of oligomeric alpha-synuclein protein beyond a single region, area, or structure in an organ. In some embodiments, the term refers to the spreading of alpha-synuclein protein to a new site or locus that is outside the single region, area, or structure and is separate from (e.g., not in physical communication with) a pre-existing site or locus that is located within the region, area, or structure. In some embodiments, the term refers to an increase in the size of a single site or locus containing alpha-synuclein protein such that the boundaries of the single site or locus fall outside of the region, area, or structure of interest.

[0120] In some embodiments, broad spreading is defined as when the size (e.g., volume) of a particular site or locus in which alpha-synuclein protein (e.g., a toxic form of alpha-synuclein protein (e.g., oligomeric alpha-synuclein protein)) is present exceeds a reference value. In some embodiments, broad spreading is defined as when the total amount of alpha-synuclein protein that is present in a region, area, or structure of an organ (e.g., a brain of a subject), or the total organ itself, is greater than a reference value. The reference value can be a specific value that is expressed in a particular unit (e.g., a unit of volume such as mm³), or more commonly, a range of values. In some embodiments, broad spreading is defined as when the size of a site or locus in which alpha-synuclein is present, or the total amount of alpha-synuclein that is present in a region, area, structure, or organ, is at least about 1.1-, 1.2-, 1.3-, 1.4-, 1.5-, 1.6-. 1.7-. 1.8-. 1.9-. 2-. 2.5-. 3-. 3.5-. 4-. 4.5-. 5-. 5.5-. 6-. 6.5-. 7-. 7.5-. 8-. 8.5-. 9-. 9.5-. 10- fold, or more greater than a reference value. The reference value can be derived from an individual before the occurrence of a disease (e.g., a neurodegenerative disease) or before the appearance of signs or symptoms of the disease, or can be derived from a population of unaffected individuals.

[0121] In some embodiments, broad spreading is associated with the onset and/or progression of one or more signs or symptoms of a disease (e.g., a neurodegenerative disease), examples of which are described herein.

[0122] A "transferrin receptor" or "TfR" as used in the context of this invention refers to transferrin receptor protein 1. The human transferrin receptor 1 polypeptide sequence is set forth in SEQ ID NO:456. Transferrin receptor protein 1 sequences from other species are also known (e.g., chimpanzee, accession number XP 003310238.1; rhesus monkey, NP_001244232.1; dog, NP_001003111.1; cattle, P_001193506.1; mouse, NP_035768.1; rat, NP_073203.1; and chicken, P_990587.1). The term "transferrin receptor" also encompasses allelic variants of exemplary reference sequences, e.g., human sequences, that are encoded by a gene at a transferrin receptor protein 1 chromosomal locus. Full-length transferrin receptor protein includes a short N-terminal intracellular region, a transmembrane region, and a large extracellular domain. The extracellular domain is characterized by three domains: a protease- like domain, a helical domain, and an apical domain.

[0123] As used herein, the term "Fc polypeptide" refers to the C-terminal region of a naturally occurring immunoglobulin heavy chain polypeptide that is characterized by an Ig fold as a structural domain. An Fc polypeptide contains constant region sequences including at least the CH2 domain and/or the CH3 domain and may contain at least part of the hinge region, but does not contain a variable region.

[0124] A "modified Fc polypeptide" refers to an Fc polypeptide that has at least one mutation, e.g., a substitution, deletion or insertion, as compared to a wild-type immunoglobulin heavy chain Fc polypeptide sequence, but retains the overall Ig fold or structure of the native Fc polypeptide.

[0125] The term "FcRn" refers to the neonatal Fc receptor. Binding of Fc polypeptides to FcRn reduces clearance and increases serum half-life of the Fc polypeptide. The human FcRn protein is a heterodimer that is composed of a protein of about 50 kDa in size that is similar to a major histocompatibility (MHC) class I protein and a 2-microglobulin of about 15 kDa in size. [0126] As used herein, an "FcRn binding site" refers to the region of an Fc polypeptide that binds to FcRn. In human IgG, the FcRn binding site, as numbered using the EU index, includes L251, M252, 1253, S254, R255, T256, M428, H433, N434, H435, and Y436. These positions correspond to positions 21 to 26, 198, and 203 to 206 of SEQ ID NO:455.

[0127] As used herein, a "native FcRn binding site" refers to a region of an Fc polypeptide that binds to FcRn and that has the same amino acid sequence as the region of a naturally occurring Fc polypeptide that binds to FcRn.

[0128] The terms "CH3 domain" and "CH2 domain" as used herein refer to immunoglobulin constant region domain polypeptides. For purposes of this application, a CH3 domain polypeptide refers to the segment of amino acids from about position 341 to about position 447 as numbered according to the EU numbering scheme, and a CH2 domain polypeptide refers to the segment of amino acids from about position 231 to about position 340 as numbered according to the EU numbering scheme and does not include hinge region sequences. CH2 and CH3 domain polypeptides may also be numbered by the IMGT (ImMunoGeneTics) numbering scheme in which the CH2 domain numbering is 1-110 and the CH3 domain numbering is 1-107, according to the IMGT Scientific chart numbering (IMGT website). CH2 and CH3 domains are part of the Fc region of an immunoglobulin. An Fc region refers to the segment of amino acids from about position 231 to about position 447 as numbered according to the EU numbering scheme, but as used herein, can include at least a part of a hinge region of an antibody. An illustrative hinge region sequence is the human IgGl hinge sequence EPKSCDKTHTCPPCP (SEQ ID NO:329).

[0129] The terms "wild-type," "native," and "naturally occurring" with respect to a CH3 or CH2 domain are used herein to refer to a domain that has a sequence that occurs in nature.

[0130] In the context of this invention, the term "mutant" with respect to a mutant polypeptide or mutant polynucleotide is used interchangeably with "variant." A variant with respect to a given wild-type CH3 or CH2 domain reference sequence can include naturally occurring allelic variants. A "non-naturally" occurring CH3 or CH2 domain refers to a variant or mutant domain that is not present in a cell in nature and that is produced by genetic modification, e.g., using genetic engineering technology or mutagenesis techniques, of a native CH3 domain or CH2 domain polynucleotide or polypeptide. A "variant" includes any domain comprising at least one amino acid mutation with respect to wild-type. Mutations may include substitutions, insertions, and deletions. [0131] The term "cross-reacts," as used herein, refers to the ability of an antibody variable region to bind to an antigen other than the antigen against which the antibody was raised. In some embodiments, cross-reactivity refers to the ability of an antibody variable region to bind to an antigen from another species than the antigen against which the antibody was raised. As a non-limiting example, an anti-alpha-synuclein antibody as described herein that is raised against a human alpha-synuclein protein can exhibit cross-reactivity with an alpha-synuclein protein from a different species (e.g., mouse or monkey).

[0132] The term "isolated," as used with reference to a nucleic acid or protein (e.g., antibody), denotes that the nucleic acid or protein is essentially free of other cellular components with which it is associated in the natural state. It is preferably in a homogeneous state. Purity and homogeneity are typically determined using analytical chemistry techniques such as electrophoresis (e.g., polyacrylamide gel electrophoresis) or chromatography (e.g., high performance liquid chromatography). In some embodiments, an isolated nucleic acid or protein (e.g., antibody) is at least 85% pure, at least 90% pure, at least 95% pure, or at least 99% pure.

[0133] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids.

[0134] Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate and O- phosphoserine. Naturally occurring a-amino acids include, without limitation, alanine (Ala), cysteine (Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (He), arginine (Arg), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gin), serine (Ser), threonine (Thr), valine (Val), tryptophan (Tip), tyrosine (Tyr), and combinations thereof. Stereoisomers of a naturally occurring a-amino acids include, without limitation, D-alanine (D-Ala), D-cysteine (D-Cys), D-aspartic acid (D-Asp), D-glutamic acid (D-Glu), D-phenylalanine (D-Phe), D-histidine (D- His), D-isoleucine (D-Ile), D-arginine (D-Arg), D-lysine (D-Lys), D-leucine (D-Leu), D- methionine (D-Met), D-asparagine (D-Asn), D-proline (D-Pro), D-glutamine (D-Gln), D- serine (D-Ser), D-threonine (D-Thr), D-valine (D-Val), D-tryptophan (D-Trp), D-tyrosine (D- Tyr), and combinations thereof. [0135] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0136] The terms "polypeptide" and "peptide" are used interchangeably herein to refer to a polymer of amino acid residues in a single chain. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymers. Amino acid polymers may comprise entirely L- amino acids, entirely D-amino acids, or a mixture of L and D amino acids.

[0137] The term "protein" as used herein refers to either a polypeptide or a dimer (i.e, two) or multimer (i.e., three or more) of single chain polypeptides. The single chain polypeptides of a protein may be joined by a covalent bond, e.g., a disulfide bond, or non-covalent interactions.

[0138] The terms "polynucleotide" and "nucleic acid" interchangeably refer to chains of nucleotides of any length, and include DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or their analogs, or any substrate that can be incorporated into a chain by a DNA or RNA polymerase. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and their analogs. Examples of polynucleotides contemplated herein include single- and double- stranded DNA, single- and double-stranded RNA, and hybrid molecules having mixtures of single- and double-stranded DNA and RNA.

[0139] The terms "conservative substitution" and "conservative mutation" refer to an alteration that results in the substitution of an amino acid with another amino acid that can be categorized as having a similar feature. Examples of categories of conservative amino acid groups defined in this manner can include: a "charged/polar group" including Glu (Glutamic acid or E), Asp (Aspartic acid orD), Asn (Asparagine orN), Gin (Glutamine or Q), Lys (Lysine or K), Arg (Arginine or R), and His (Histidine or H); an "aromatic group" including Phe (Phenylalanine or F), Tyr (Tyrosine or Y), Trp (Tryptophan or W), and (Histidine or H); and an "aliphatic group" including Gly (Glycine or G), Ala (Alanine or A), Val (Valine or V), Leu (Leucine or L), He (Isoleucine or I), Met (Methionine or M), Ser (Serine or S), Thr (Threonine or T), and Cys (Cysteine or C). Within each group, subgroups can also be identified. For example, the group of charged or polar amino acids can be sub-divided into sub-groups including: a "positively-charged sub-group" comprising Lys, Arg and His; a "negatively- charged sub-group" comprising Glu and Asp; and a "polar sub-group" comprising Asn and Gin. In another example, the aromatic or cyclic group can be sub-divided into sub-groups including: a "nitrogen ring sub-group" comprising Pro, His and Tip; and a "phenyl sub-group" comprising Phe and Tyr. In another further example, the aliphatic group can be sub-divided into sub-groups, e.g., an "aliphatic non-polar sub-group" comprising Val, Leu, Gly, and Ala; and an "aliphatic slightly-polar sub-group" comprising Met, Ser, Thr, and Cys. Examples of categories of conservative mutations include amino acid substitutions of amino acids within the sub-groups above, such as, but not limited to: Lys for Arg or vice versa, such that a positive charge can be maintained; Glu for Asp or vice versa, such that a negative charge can be maintained; Ser for Thr or vice versa, such that a free -OH can be maintained; and Gin for Asn or vice versa, such that a free - H2 can be maintained. In some embodiments, hydrophobic amino acids are substituted for naturally occurring hydrophobic amino acid, e.g., in the active site, to preserve hydrophobicity.

[0140] The terms "identical" or percent "identity," in the context of two or more polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues, e.g., at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or greater, that are identical over a specified region when compared and aligned for maximum correspondence over a comparison window or designated region, as measured using a sequence comparison algorithm or by manual alignment and visual inspection.

[0141] For sequence comparison of polypeptides, typically one amino acid sequence acts as a reference sequence, to which a candidate sequence is compared. Alignment can be performed using various methods available to one of skill in the art, e.g., visual alignment or using publicly available software using known algorithms to achieve maximal alignment. Such programs include the BLAST programs, ALIGN, ALIGN-2 (Genentech, South San Francisco, Calif.) or Megalign (DNASTAR). The parameters employed for an alignment to achieve maximal alignment can be determined by one of skill in the art. For sequence comparison of polypeptide sequences for purposes of this application, the BLASTP algorithm standard protein BLAST for aligning two proteins sequence with the default parameters is used.

[0142] The terms "corresponding to," "determined with reference to," or "numbered with reference to" when used in the context of the identification of a given amino acid residue in a polypeptide sequence, refers to the position of the residue of a specified reference sequence when the given amino acid sequence is maximally aligned and compared to the reference sequence. Thus, for example, an amino acid residue in a modified Fc polypeptide "corresponds to" an amino acid in SEQ ID NO:455, when the residue aligns with the amino acid in SEQ ID NO:455 when optimally aligned to SEQ ID NO:455. The polypeptide that is aligned to the reference sequence need not be the same length as the reference sequence.

[0143] The term "subject," "individual," and "patient," as used interchangeably herein, refer to a mammal, including but not limited to humans, non-human primates, rodents (e.g., rats, mice, and guinea pigs), rabbits, cows, pigs, horses, and other mammalian species. In one embodiment, the patient is a human.

[0144] The terms "treatment," "treating," and the like are used herein to generally mean obtaining a desired pharmacologic and/or physiologic effect. "Treating" or "treatment" may refer to any indicia of success in the treatment or amelioration of a neurodegenerative disease (e.g., a synucleinopathy), including any objective or subjective parameter such as abatement, remission, improvement in patient survival, increase in survival time or rate, diminishing of symptoms or making the disease more tolerable to the patient, slowing in the rate of degeneration or decline, or improving a patient' s physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters. The effect of treatment can be compared to an individual or pool of individuals not receiving the treatment, or to the same patient prior to treatment or at a different time during treatment.

[0145] As used herein, the term "synucleinopathy" refers to a pathological condition (e.g., a neurodegenerative disease) that is characterized by an abnormal accumulation of alpha- synuclein protein in neural tissue (e.g., brain tissue). Examples of synucleinopathies include Parkinson' s disease, dementia with Lewy bodies, and multiple system atrophy. Synucleinopathies are typically characterized by the presence of Lewy bodies or Lewy neurites, which contain significant amounts of alpha-synuclein aggregates, including alpha-synuclein oligomers and/or fibrils. In some instances, synucleinopathies are characterized by the spreading of oligomers and/or fibrils of alpha-synuclein protein throughout the diseased tissue. The accumulation of alpha-synuclein protein results in, for example, mitochondrial dysfunction, ER-Golgi traffic dysfunction, inhibition of autophagy and proteasome pathways, and disruption of synaptic tissue, all of which can lead to cell death. [0146] The term "pharmaceutically acceptable excipient" refers to a non-active pharmaceutical ingredient that is biologically or pharmacologically compatible for use in humans or animals, such as but not limited to a buffer, carrier, or preservative.

[0147] As used herein, a "therapeutic amount" or "therapeutically effective amount" of an agent (e.g., an antibody as described herein) is an amount of the agent that treats, alleviates, abates, or reduces the severity of symptoms of a disease in a subject. A "therapeutic amount" or "therapeutically effective amount" of an agent (e.g., an antibody as described herein) may improve patient survival, increase survival time or rate, diminish symptoms, make an injury, disease, or condition (e.g., a neurodegenerative disease) more tolerable, slow the rate of degeneration or decline, or improve a patient' s physical or mental well-being.

[0148] The term "administer" refers to a method of delivering agents, compounds, or compositions to the desired site of biological action. These methods include, but are not limited to, topical delivery, parenteral delivery, intravenous delivery, intradermal delivery, intramuscular delivery, intrathecal delivery, colonic delivery, rectal delivery, or intraperitoneal delivery. In one embodiment, an antibody as described herein is administered intravenously.

III. ANTI- ALPHA-SYNUCLEIN ANTIBODIES

[0149] In one aspect, antibodies and antigen-binding portions of antibodies that specifically bind to an alpha-synuclein protein (e.g., human alpha-synuclein) are provided. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to monomeric and/or oligomeric alpha-synuclein. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to monomeric alpha-synuclein. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to oligomeric alpha-synuclein. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to monomeric and oligomeric alpha-synuclein. In some embodiments, the antibody or antigen- binding portion thereof has a higher binding affinity for oligomeric human alpha-synuclein protein than for monomeric human alpha-synuclein protein. In some embodiments, the antibody or antigen-binding portion thereof has a higher binding affinity for monomeric human alpha-synuclein protein than for oligomeric human alpha-synuclein protein.

[0150] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to soluble alpha-synuclein protein (e.g., human alpha-synuclein protein) and/or alpha- synuclein protein fibrils. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to soluble alpha-synuclein. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to alpha-synuclein fibrils. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to soluble alpha-synuclein and alpha-synuclein fibrils.

[0151] In some embodiments, the antibody is a pan-alpha-synuclein antibody or antigen- binding portion thereof. In some embodiments, the pan-alpha-synuclein antibody or antigen- binding portion thereof specifically binds to monomeric human alpha-synuclein protein, oligomeric human alpha-synuclein protein, soluble human alpha-synuclein protein, and human alpha-synuclein protein fibrils.

[0152] In some embodiments, the antibody specifically binds to one or more splice isoforms of human alpha-synuclein protein, non-limiting examples of which include full-length alpha- synuclein (SEQ ID NO: l) and isoforms resulting from alternative splicing (e.g., alpha- synuclein-1 12 and alpha-synuclein-126).

[0153] In another aspect, antibodies and antigen-binding portions of antibodies that neutralize alpha-synuclein protein (e.g., human alpha-synuclein) are provided. In some embodiments, the antibody or antigen-binding portion thereof neutralizes human alpha- synuclein protein in a subject, e.g., in a brain of a subject. In some embodiments, the antibody or antigen-binding portion thereof neutralizes alpha-synuclein protein by preventing, reducing, or inhibiting alpha-synuclein oligomerization and/or aggregation. In some embodiments, the antibody of antigen-binding portion thereof prevents, reduces, or inhibits alpha-synuclein oligomerization and/or aggregation in a subject, e.g., in a brain of a subject. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits alpha-synuclein oligomerization. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits alpha-synuclein aggregation. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits alpha-synuclein oligomerization and aggregation.

[0154] In some embodiments, an anti-alpha-synuclein antibody prevents, reduces, or inhibits alpha-synuclein spreading by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% (e.g., compared to the amount of alpha- synuclein spreading in the absence of the anti-alpha-synuclein antibody). In some embodiments, an anti-alpha-synuclein antibody prevents, reduces, or inhibits alpha-synuclein oligomerization by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% (e.g., compared to the amount of alpha- synuclein oligomerization in the absence of the anti-alpha-synuclein antibody). In some embodiments, an anti-alpha-synuclein antibody prevents, reduces, or inhibits alpha-synuclein aggregation by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% (e.g., compared to the amount of alpha-synuclein aggregation in the absence of the anti-alpha-synuclein antibody). Methods for measuring alpha-synuclein seeding, spreading, oligomerization, and/or aggregation are known in the art.

[0155] In some embodiments, the antibody or antigen-binding portion thereof neutralizes alpha-synuclein protein by preventing, reducing, or inhibiting the spreading of alpha-synuclein protein oligomers and/or fibrils. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits the spreading of alpha-synuclein protein oligomers and/or fibrils in a subject, e.g., in the brain of a subject. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits the spreading of alpha-synuclein protein oligomers. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits the spreading of alpha-synuclein protein fibrils. In some embodiments, the antibody or antigen-binding portion thereof prevents, reduces, or inhibits the spreading of alpha-synuclein protein oligomers and fibrils.

[0156] In some embodiments, the antibody or antigen-binding portion thereof neutralizes monomeric human alpha-synuclein, oligomeric human alpha-synuclein, soluble human alpha- synuclein, and/or human alpha-synuclein fibrils. In some embodiments, the antibody or antigen-binding portion thereof that neutralizes alpha-synuclein is a pan-alpha-synuclein antibody or a multispecific (e.g., bispecific) antibody.

[0157] In another aspect, antibodies and antigen-binding portions of antibodies that bind to alpha-synuclein protein (e.g., human alpha-synuclein) and thereby prevent or treat a neurodegenerative disease (e.g., a synucleinopathy or tauopathy) in a subject (e.g., a human subject) are provided. In some embodiments, the antibody or antigen-biding portion thereof prevents or treats a neurodegenerative disease that is characterized by the presence of Lewy bodies and/or Lewy neurites. In some embodiments, the antibody or antigen-binding portion thereof prevents or treats a neurodegenerative disease that is characterized by the presence of alpha-synuclein accumulation or aggregation (e.g., alpha-synuclein oligomers and/or fibrils).

[0158] In some embodiments, the antibody or antigen-binding portion prevents or treats a neurodegenerative disease that is selected from the group consisting of Parkinson' s disease, dementia with Lewy bodies, multiple system atrophy, Huntington' s disease, Alzheimer's disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down's syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden-Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle-predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick's disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia. In some embodiments, the antibody or antigen-binding portion thereof prevents or treats a synucleinopathy. In some embodiments, the synucleinopathy is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy.

[0159] In some embodiments, the antibody or antigen-binding portion thereof slows the progression of symptoms of a neurodegenerative disease (e.g., in a subject). In some embodiments, the antibody or antigen-binding portion thereof causes the reversal of symptoms of a neurodegenerative disease. As non-limiting examples, symptoms of a neurodegenerative disease include memory deficits, cognitive deficits, motor deficits, sensory deficits, and speech deficits.

Binding characteristics of anti-alpha-synuclein antibodies

[0160] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof specifically binds to full-length human alpha-synuclein (SEQ ID NO: l) with high affinity. In some embodiments, the antibody or antigen-binding portion thereof has a binding affinity (KD) for SEQ ID NO: l of less than about 250 nM, e.g., less than about 240 nM, less than about 230 nM, less than about 220 nM, less than about 210 nM, less than about 200 nM, less than about 190 nM, less than about 180 nM, less than about 170 nM, less than about 160 nM, less than about 150 nM, less than about 140 nM, less than about 130 nM, less than about 120 nM, less than about 110 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 25 nM, less than about 20 nM, less than about 15 nM, less than about 10 nM, less than about 5 nM, less than about 1 nM, less than about 800 pM, less than about 500 pM, less than about 250 pM, less than about 150 pM, less than about 100 pM, less than about 50 pM, less than about 40 pM, less than about 30 pM, less than about 20 pM, less than about 10 pM, or less than about 5 pM. In some embodiments, the antibody has a KD for SEQ ID NO: 1 in the range of about 1 pM to about 50 nM, e.g., about 1 pM to about 25 nM, about 1 pM to about 20 nM, about 1 pM to about 10 nM, about 1 pM to about 5 nM, about 1 pM to about 1 nM, about 1 pM to about 800 pM, about 1 pM to about 700 pM, about 1 pM to about 600 pM about 1 pM to about 500 pM, about 5 pM to about 250 pM, about 10 pM to about 800 pM, about 10 pM to about 500 pM, or about 10 pM to about 100 pM.

[0161] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof specifically binds to more than one splice isoform of human alpha-synuclein (e.g., two or three splice isoforms such as "full-length" alpha-synuclein (SEQ ID NO: 1), alpha-synuclein- 1 12, or alpha-synuclein- 126) with high affinity. In some embodiments, the antibody or antigen-binding portion thereof has a KD for two or more isoforms of human alpha-synuclein of less than about 50 nM, e.g., less than about 40 nM, less than about 30 nM, less than about 25 nM, less than about 20 nM, less than about 15 nM, less than about 10 nM, less than about 5 nM, less than about 1 nM, less than about 800 pM, less than about 500 pM, less than about 250 pM, less than about 150 pM, less than about 100 pM, less than about 50 pM, less than about 40 pM, less than about 30 pM, less than about 20 pM, or less than about 10 pM. In some embodiments, the antibody has a KD for two or more isoforms of human alpha-synuclein in the range of about 1 pM to about 50 nM, e.g., about 1 pM to about 25 nM, about 1 pM to about 20 nM, about 1 pM to about 10 nM, about 1 pM to about 5 nM, about 1 pM to about 1 nM, about 1 pM to about 800 pM, about 1 pM to about 700 pM, about 1 pM to about 600 pM about 1 pM to about 500 pM, about 5 pM to about 250 pM, about 10 pM to about 800 pM, about 10 pM to about 500 pM, or about 10 pM to about 100 pM.

[0162] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof exhibits cross-reactivity with cynomolgus monkey ("cyno") alpha-synuclein. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof exhibits cross-reactivity with mouse alpha-synuclein. In some embodiments, an anti-alpha-synuclein antibody or an antigen-binding portion thereof exhibits cross-reactivity with cynomolgus monkey alpha-synuclein and mouse alpha-synuclein. [0163] Methods for analyzing binding affinity, binding kinetics, and cross-reactivity are known in the art. These methods include, but are not limited to, solid-phase binding assays (e.g., ELISA assay), immunoprecipitation, surface plasmon resonance (e.g., Biacore™ (GE Healthcare, Piscataway, NJ)), kinetic exclusion assays (e.g., KinExA®), flow cytometry, fluorescence-activated cell sorting (FACS), BioLayer interferometry (e.g., Octet™ (ForteBio, Inc., Menlo Park, CA)), and Western blot analysis. In some embodiments, ELISA is used to determine binding affinity and/or cross-reactivity. Methods for performing ELISA assays are known in the art, and are also described in the Examples section below. In some embodiments, surface plasmon resonance (SPR) is used to determine binding affinity, binding kinetics, and/or cross-reactivity. In some embodiments, kinetic exclusion assays are used to determine binding affinity, binding kinetics, and/or cross-reactivity. In some embodiments, BioLayer interferometry assays are used to determine binding affinity, binding kinetics, and/or cross- reactivity.

[0164] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence GAPQEGILEDMPVDP (SEQ ID NO:256), which corresponds to residues 106-120 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 106-120 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 106-120 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 contiguous amino acids within residues 106-120 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5- 15, 5-12, 5-10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 106-120 of SEQ ID NO: l .

[0165] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence GILEDMPVDPDNEAY (SEQ ID NO:257), which corresponds to residues 1 1 1-125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 1 1 1-125 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 1 1 1-125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 contiguous amino acids within residues 1 1 1-125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5- 15, 5-12, 5-10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 1 1 1-125 of SEQ ID NO: l .

[0166] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 106-125 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 106-120 of SEQ ID NO: l or recognizes an epitope within residues 1 1 1-125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 106-125 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 106-120 of SEQ ID NO: l or recognizes an epitope comprising residues 1 1 1-125 of SEQ ID NO: 1.

[0167] In some embodiments, the antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 106-120 or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 1 1 1-125 of SEQ ID NO: l .

[0168] In some embodiments, the anti-alpha-synuclein antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 106-120 or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 1 1 1-125 of SEQ ID NO: l . [0169] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence VTGVTAVAQKTVEGA (SEQ ID NO:261), which corresponds to residues 71-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 71-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 amino acids within residues 71-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 contiguous amino acids within residues 71-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5-10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 71-85 of SEQ ID NO: l .

[0170] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence GSIAAATGFVKKDQL (SEQ ID NO:258), which corresponds to residues 86-100 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 86-100 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 amino acids within residues 86-100 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14, contiguous amino acids within residues 86-100 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5-10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 86-105 of SEQ ID NO: l .

[0171] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence ATGFVKKDQLGKNEE (SEQ ID NO:259), which corresponds to residues 91-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 91-105 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 91-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, at least 14 contiguous amino acids within residues 91-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5- 10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 91-105 of SEQ ID NO: 1.

[0172] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 71-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 71-85, residues 86-100, or residues 91-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 71-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 71-85, residues 86-100, or residues 91-105 of SEQ ID NO: 1.

[0173] In some embodiments, the antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 71-85, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 86-100, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 91- 105 of SEQ ID NO: 1.

[0174] In some embodiments, the anti-alpha-synuclein antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 71-85, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 86-100, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least

13, or at least 14) contiguous amino acids within residues 91-105 of SEQ ID NO: l .

[0175] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence VGGAVVTGVTAVAQK (SEQ ID NO:260), which corresponds to residues 66-80 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 66-80 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 66-80 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, at least 14 contiguous amino acids within residues 66-80 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5- 10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 66-80 of SEQ ID NO: 1.

[0176] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 66-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 66-80 or residues 71-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 66-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 66-80 of SEQ ID NO: l or recognizes an epitope comprising residues 71-85 of SEQ ID NO: l .

[0177] In some embodiments, the antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 66-80 or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 71-85 of SEQ ID NO: l . [0178] In some embodiments, the anti-alpha-synuclein antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 66-80 or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 71-85 of SEQ ID NO: l .

[0179] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence TVEGAGSIAAATGFV (SEQ ID NO:262), which corresponds to residues 81-95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 81-95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 81-95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, at least 14 contiguous amino acids within residues 81-95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5- 10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 81-95 of SEQ ID NCv l .

[0180] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence GK EEGAPQEGILED (SEQ ID NO:265), which corresponds to residues 101-1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 101-1 15 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 101-1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, at least 14 contiguous amino acids within residues 101-1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5- 10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 101-115 of SEQ ID NO: 1.

[0181] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 81-1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 81-95, residues 86- 100, residues 91-105, or residues 101-1 15 of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 81-1 15 of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 81-95, residues 86-100, residues 91-105, or residues 101- 1 15 of SEQ ID NCv l .

[0182] In some embodiments, the antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 81-95, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 86-100, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 91-105, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 101-1 15 of SEQ ID NO: 1.

[0183] In some embodiments, the anti-alpha-synuclein antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 81-95, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 86-100, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 91-105, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 101-1 15 of SEQ ID NO: 1.

[0184] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence GVLYVGSKTKEGVVH (SEQ ID NO:263), which corresponds to residues 36-50 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 36-50 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 amino acids within residues 36-50 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 contiguous amino acids within residues 36-50 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5-10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 36-50 of SEQ ID NO: 1.

[0185] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence KKDQLGKNEEGAPQE (SEQ ID NO:264), which corresponds to residues 96-110 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 96-110 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 amino acids within residues 96-110 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 contiguous amino acids within residues 96-110 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5-10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 96-110 of SEQ ID NO: 1.

[0186] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 36-50 of SEQ ID NO: 1 and recognizes an epitope within residues 86-115 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 36-50, 86-100, 96-110, or 101-115 of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 36-50 of SEQ ID NO: l and recognizes an epitope comprising residues 86-1 15 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 36-50, 86-100, 96-1 10, or 101-1 15 of SEQ ID NCv l .

[0187] In some embodiments, the antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 36-50, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 86-100, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 96- 1 10, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least

1 1, at least 12, at least 13, or at least 14) amino acids within residues 101-1 15 of SEQ ID NO: 1.

[0188] In some embodiments, the anti-alpha-synuclein antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 36-50, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least

12, at least 13, or at least 14) contiguous amino acids within residues 86-100, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 96-1 10, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 101-1 15 of SEQ ID NO: 1.

[0189] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence DNEAYEMPSEEGYQD (SEQ ID NO:323), which corresponds to residues 121-135 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 121-135 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14 amino acids within residues 121-135 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14 contiguous amino acids within residues 121-135 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5- 10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 121-135 of SEQ ID NO: 1.

[0190] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence EMPSEEGYQDYEPEA (SEQ ID NO:322), which corresponds to residues 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 126-140 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 amino acids within residues 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14 contiguous amino acids within residues 126-140 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope comprising 4-15, 4-12, 4-10, 4-8, 5-15, 5-12, 5- 10, 5-8, 6-15, 6-12, 6-10, 6-8, 8-15, 8-12, 8-10, 10-15, 10-12, or 12-15 contiguous amino acids within residues 126-140 of SEQ ID NO: l .

[0191] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein within residues 86-105 of SEQ ID NO: l or recognizes an epitope within residues 121-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 86-100, 91-105, 121-135, or 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 86-105 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 86-105 and 121-140 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope within residues 121-140 of SEQ ID NO: l . [0192] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein comprising residues 86-105 of SEQ ID NO: l or recognizes an epitope comprising residues 121-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising residues 86-100, 91-105, 121-135, or 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising residues 86-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising residues 86-105 and 121 -140 of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising residues 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising residues 121-140 of SEQ ID NCv l .

[0193] In some embodiments, the antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 86-100, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 91-105, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) amino acids within residues 121- 135, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least

1 1, at least 12, at least 13, or at least 14) amino acids within residues 126-140 of SEQ ID NO: l .

[0194] In some embodiments, the anti-alpha-synuclein antibody recognizes an epitope comprising at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 86-100, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least

12, at least 13, or at least 14) contiguous amino acids within residues 91-105, at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 121-135, or at least 4 (e.g., at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, or at least 14) contiguous amino acids within residues 126-140 of SEQ ID NO: l .

[0195] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope of human alpha-synuclein having or consisting of the sequence EQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGK EEGAPQEGILED MP VDPD E A YEMP SEEG YQD YEPE A (SEQ ID NO:321), which corresponds to residues 61-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen- binding portion thereof recognizes an epitope within residues 61-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 1 1, at least 12, at least 13, at least 14, at least 15, at least 16, a least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, or at least 79 amino acids within residues 61-140 of SEQ ID NO: l .

[0196] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, a least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, or at least 79 contiguous amino acids within residues 61-140 of SEQ ID NO: 1.

[0197] In some embodiments, an anti-alpha-synuclein antibody or antigen-binding portion thereof recognizes an epitope comprising 5-10, 5-15, 5-20, 5-25, 5-30, 5-35, 5-40, 5-45, 5-50, 5-55, 5-60, 5-65, 5-70, 5-75, 5-80, 10-15, 10-20, 10-25, 10-30, 10-35, 10-40, 10-45, 10-50, 10- 55, 10-60, 10-65, 10-70, 10-75, 10-80, 15-20, 15-25, 15-30, 15-35, 15-40, 15-45, 15-50, 15- 55, 15-60, 15-65, 15-70, 15-75, 15-80, 20-25, 20-30, 20-35, 20-40, 20-45, 20-50, 20-55, 20- 60, 20-65, 20-70, 20-75, 20-80, 25-30, 25-35, 25-40, 25-45, 25-50, 25-55, 25-60, 25-65, 25- 70, 25-75, 25-80, 30-35, 30-40, 30-45, 30-50, 30-55, 30-60, 30-65, 30-70, 30-75, 30-80, 35- 40, 35-45, 35-50, 35-55, 35-60, 35-65, 35-70, 35-75, 35-80, 40-45, 40-50, 40-55, 40-60, 40- 65, 40-70, 40-75, 40-80, 45-50, 45-55, 45-60, 45-65, 45-70, 45-80, 50-55, 50-60, 50-65, 50- 70, 50-75, 50-80, 55-60, 55-65, 55-70, 55-75, 55-80, 60-65, 60-70, 60-75, 60-80, 65-70, 65- 75, 65-80, 70-75, 70-80, or 75-80 contiguous amino acids within residues 61-140 of SEQ ID NO: l .

Anti-alpha-synuclein antibody sequences

[0198] In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein comprises a light chain sequence, or a portion thereof, and/or a heavy chain sequence, or a portion thereof, derived from any of the following antibodies described herein: Clone 1F06, Clone 1F06 C99M, Clone 3E02, Clone 3F02, Clone 2D01, Clone 2E01, Clone 2F01, Clone 1F02, Clone 2A02, Clone 1H02, Clone 2B02, Clone 2C02, Clone 2D02, Clone 1A03, Clone 2A03, Clone 2G02, Clone 1D01, Clone 2C03, Clone 1D02, Clone 2A04, Clone 2B04, Clone 2C04, Clone 2E03, Clone 2E04, Clone 2G04, Clone 2H04, Clone 2B05, Clone 2C05, Clone 2D05, Clone 2E05, Clone 2G05, Clone 2A06, Clone 2H05, Clone 2C06, Clone 2G06, Clone 2H06, Clone 1D03, Clone 2A07, Clone 2B07, Clone 1E03, Clone 1F03, Clone 1G03, Clone 2A08, Clone 2B08, Clone 2C07, Clone 2D07, Clone 2G07, Clone 2H07, Clone 2D08, Clone 2B09, Clone 2C09, Clone 2D09, Clone 2F08, Clone 2G08, Clone 2H08, Clone 2E09, Clone 2F09, Clone 1E04, Clone 2G09, Clone 2F 10, Clone 1G06, Clone 3C02, Clone 3D02, Clone 4F5, Clone 1G4, Clone 16D4, Clone 28B6, Clone 3E12, Clone 3D5, Clone 46D2, Clone 1F4, and Clone 1F4 (C^S variant). The amino acid sequences of the light chain variable region (VL) and heavy chain variable region (VH) of the anti-alpha-synuclein antibodies described herein are set forth in Table 15 below.

[0199] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:2-61, 266, and 269-275. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:2-61, 266, and 269-275. In some embodiments, a heavy chain variable region sequence having at least 90% sequence identity to a reference sequence (e.g., SEQ ID NOs:2-61, 266, and 269-275) contains one, two, three, four, five, six, seven, eight, nine, ten or more substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence but retains the ability to specifically bind to a human alpha-synuclein protein and recognize one or more epitopes as described herein. In some embodiments, a heavy chain variable region contains one, two, or three substitutions (e.g., conservative substitutions) in any one of SEQ ID NOs:2-61, 266, and 269-275.

[0200] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 146-204 and 292-299. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs: 146-204 and 292-299. In some embodiments, a light chain variable region sequence having at least 90% sequence identity to a reference sequence (e.g., SEQ ID NOs: 146-204 and 292-299) contains one, two, three, four, five, six, seven, eight, nine, ten or more substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence but retains the ability to bind to human alpha-synuclein and recognize one or more epitopes as described herein. In some embodiments, a light chain variable region contains one, two, or three substitutions (e.g., conservative substitutions) in any one of SEQ ID NOs: 146-204 and 292-299.

[0201] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:2-61, 266, and 269-275 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 146-204 and 292-299. In some embodiments, an anti- alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:2-61, 266, and 269-275 and further comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs: 146-204 and 292-299. Anti-alpha-synuclein antibodies that recognize an epitope within residues 106-125

[0202] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 106-125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 106-120 of SEQ ID NO: l or an epitope within residues 1 1 1-125 of SEQ ID NO: 1. In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0203] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues G106, A107, P108, Q109, El 10, Gi l l, 11 12, LI 13, El 14, D1 15, Ml 16, PI 17, VI 18, D 1 19, P120, D121, N122, E123, A124, or Y125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, or all 20 of the following residues G106, A107, P108, Q109, El 10, Gi l l, 11 12, L1 13, E1 14, D1 15, M1 16, P1 17, V1 18, D1 19, P120, D121, N122, E123, A124, or Y125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to non-contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: l .

[0204] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G106 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A107 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 08 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q 109 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue El 10 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Gi l l of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue II 12 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue LI 13 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 14 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Ml 16 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 17 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue VI 18 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D1 19 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 20 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D121 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N122 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E123 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A124 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y125 of SEQ ID NO: 1.

[0205] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GAPQEGILEDMPVDP (SEQ ID NO:256), which corresponds to residues 106-120 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:256 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0206] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GILEDMPVDPDNEAY (SEQ ID NO:257), which corresponds to residues 1 1 1-125 of SEQ ID NO: 1. In some embodiments, the peptide sequence set forth in SEQ ID NO:257 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0207] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240, or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268; (d) a light chain CDRl having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:205 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:205; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:231 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:231.

[0208] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDRl of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDRl of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0209] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:205; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

[0210] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDRl of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDRl of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0211] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:2-3 and 266. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 146-148. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:2-3 and 266, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 146-148.

[0212] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:2-3 and 266. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs: 146-148. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:2-3 and 266, and a light chain variable region comprising the amino acid of any one of SEQ ID NOs: 146-148.

[0213] In some embodiments, an anti-alpha-synuclein antibody comprises one or more sequences that are variants of one or more consensus sequences or are encompassed by one or more consensus sequences. Consensus sequences can be identified by aligning heavy chain or light chain sequences (e.g., CDRs) for antibodies that bind to the same or similar (e.g., overlapping) epitopes to determine conserved amino acids or motifs (i.e., where alteration in sequences may alter protein function) and regions where variation occurs in alignment of sequences (i.e., where variation of sequence is not likely to significantly affect protein function). In some embodiments, one or more consensus sequences can be identified for antibodies that recognize the same or similar epitope as 1F06. Exemplary 1F06-Iike consensus sequences include SEQ ID NOs:240 and 244. In the consensus sequences of SEQ ID NOs:240 and 244, the capitalized letter represents an amino acid residue that is absolutely conserved among the aligned sequences (e.g., aligned CDR sequences), while "x" represents an amino acid residue that is not absolutely conserved among the aligned sequences. While "x" can be any amino acid, it will be appreciated that when selecting an amino acid to insert at a position marked by an "x" that in some embodiments, the amino acid is selected from those amino acids found at the corresponding position in the aligned sequences.

[0214] In some embodiments, the antibody comprises a heavy chain CDRl sequence having the consensus sequence GFxFSDYGMH (SEQ ID NO:240). In some embodiments, the heavy chain CDRl consensus sequence comprises the sequence GF[I/T]FSDYGMH (SEQ ID NO:622).

[0215] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence IS SGNxT VYYADTVKx (SEQ ID NO:244). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence ISSGN[N/D]TVYYADTVK[G/D] (SEQ ID NO:623).

[0216] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 106-125 of SEQ ID NO: 1 comprises a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:240. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:62 and 63.

[0217] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 106-125 of SEQ ID NO: 1 comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:244. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:81 and 82.

[0218] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 106-125 of SEQ ID NO: 1 comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:268. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 121 and 267.

[0219] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 106-125 of SEQ ID NO: 1 comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:240; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:244; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:268; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:205; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

[0220] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 106-125 of SEQ ID NO: 1 comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:62 and 63; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:81 and 82; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 121 and 267; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

1F06

[0221] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:62, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:81, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:231. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:62, 81, 121, 205, 223, and 231, respectively.

[0222] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:2. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2.

[0223] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 147. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147.

[0224] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:2 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 147. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147.

[0225] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:62, 81, 121, 205, 223, and 231, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147).

1F06 C99M variant

[0226] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:81, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:267. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:231. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:62, 81, 267, 205, 223, and 231, respectively.

[0227] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:266. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:266.

[0228] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 147. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147.

[0229] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:266 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 147. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:266 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147.

[0230] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:62, 81, 267, 205, 223, and 231, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:266 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147).

3E02

[0231] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:62, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:81, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:231. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:62, 81, 121, 205, 223, and 231, respectively.

[0232] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:2. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2.

[0233] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 148. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 148.

[0234] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:2 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 148. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 148.

[0235] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:62, 81, 121, 205, 223, and 231, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 148). 3F02

[0236] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:63, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:82, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 121. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:231. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 82, 121, 205, 223, and 231, respectively.

[0237] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99%> sequence identity) to SEQ ID NO:3. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3.

[0238] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90%> sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 146. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 146.

[0239] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90%> sequence identity (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%>, or at least 99%> sequence identity) to SEQ ID NO:3 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90%> sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 146. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 146.

[0240] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 82, 121, 205, 223, and 231, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 146).

Anti-alpha-synuclein antibodies that recognize an epitope within residues 71-105

[0241] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 71-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 71-85 of SEQ ID NO: l, recognizes an epitope within residues 86-100 of SEQ ID NO: l, or recognizes an epitope within residues 91-105 of SEQ ID NO: l . In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0242] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues V71, T72, G73, V74, T75, A76, V77, A78, Q79, K80, T81, V82, E83, G84, A85, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, or E105 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or all 35 of the following residues V71, T72, G73, V74, T75, A76, V77, A78, Q79, K80, T81, V82, E83, G84, A85, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, or E105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to non-contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: 1.

[0243] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V71 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T72 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G73 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V74 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue T75 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A76 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V77 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A78 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q79 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K80 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T81 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V82 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue E83 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G84 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A85 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G86 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue S87 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue 188 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A89 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A90 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A91 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T92 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G93 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue F94 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K96 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K97 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D98 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue Q99 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue L100 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G101 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K 102 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N103 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E104 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E105 of SEQ ID NO: 1.

[0244] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence VTGVTAVAQKTVEGA (SEQ ID NO:261), which corresponds to residues 71-85 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:261 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0245] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GSIAAATGFVKKDQL (SEQ ID NO:258), which corresponds to residues 86-100 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:258 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0246] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence ATGFVKKDQLGKNEE (SEQ ID NO:259), which corresponds to residues 91-105 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:259 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0247] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:83-1 15, 245, and 246 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:83-1 15, 245, and 246; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:206-220 and 251 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:206- 220 and 251; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:224-228 and 253 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:224-228 and 253; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:232-237 and 255 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:232-237 and 255.

[0248] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0249] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:83-l 15, 245, and 246; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 122- 141 and 249; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206-220 and 251; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:224-228 and 253; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232-237 and 255.

[0250] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0251] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:4-55. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 149-197. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:4-55, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 149- 197.

[0252] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:4-55. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs: 149-197. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:4-55, and a light chain variable region comprising the amino acid of any one of SEQ ID NOs: 149- 197.

[0253] In some embodiments, an anti-alpha-synuclein antibody comprises one or more sequences that are variants of one or more consensus sequences or are encompassed by one or more consensus sequences. In some embodiments, one or more consensus sequences can be identified for antibodies that recognize the same or similar epitope as 2D02. Exemplary 2D02- like consensus sequences include SEQ ID NOs:241, 242, 245, 246, 249, 251, 253, 255, 317, and 318. In the consensus sequences of SEQ ID NOs:241, 242, 245, 246, 249, 251, 253, 255, 317, and 318, the capitalized letter represents an amino acid residue that is absolutely conserved among the aligned sequences (e.g., aligned CDR sequences), while "x" represents an amino acid residue that is not absolutely conserved among the aligned sequences. While "x" can be any amino acid, it will be appreciated that when selecting an amino acid to insert at a position marked by an "x" that in some embodiments, the amino acid is selected from those amino acids found at the corresponding position in the aligned sequences.

[0254] In some embodiments, the antibody comprises a heavy chain CDR1 sequence having the consensus sequence xYxIxxxxY (SEQ ID NO:241). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence

[G/D]Y[P/S]I[T/P][S/R/N][G/Y/N/F][Y/F]Y (SEQ ID NO:624). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence GYSITxxYY (SEQ ID NO:625).

[0255] In some embodiments, the antibody comprises a heavy chain CDR1 sequence having the consensus sequence GxTFTDYYxN (SEQ ID NO:242). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence G[H/Y/F/N]TFTDYY[M/I]N (SEQ ID NO:626).

[0256] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence IxxDGxxxYxPSLxx (SEQ ID NO:245). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence I[G/N/S/T][D/Y/S/Q/T]DG[S/Y/N/T][N/S/D/T/H][N/D/S]Y[N/T/S]PSL[K/E][N/D] (SEQ ID NO:627). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence IxxDGxxxYxPSLKN (SEQ ID NO:628).

[0257] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence IxPxxGxxxxxQxFxx (SEQ ID NO:246). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence I[H/N]P[Y/N/H][T/S/R/N]G[D/E/G][T/M][R/N/Q/S][Y/N][N/S/T/A]

(SEQ ID NO:629) In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence IHPYxGxxxxxQxFKG (SEQ ID NO:630).

[0258] In some embodiments, the antibody comprises a heavy chain CDR3 sequence having the consensus sequence xxGDxx (SEQ ID NO:249). In some embodiments, the heavy chain CDR3 consensus sequence comprises the sequence

[A/I/L/T/V][R/S/K/T]GD[NA^/S/L/D/G/I][Y/V/N/D/H/L/A] (SEQ ID NO:631). In some embodiments, the heavy chain CDR3 consensus sequence comprises the sequence xxGDVx (SEQ ID NO:632). [0259] In some embodiments, the antibody comprises a light chain CDRl sequence having the consensus sequence xxxxSLLxxDGxTYLN (SEQ ID NO:251). In some embodiments, the light chain CDRl consensus sequence comprises the sequence [K/R][S/T][S/N][Q/R/K/L]SLL[D/Y][S/H/I/RA^r/N/K]DG[E/K]TYLN (SEQ ID NO:633). In some embodiments, the light chain CDRl consensus sequence comprises the sequence KSSxSLLxxDGETYLN (SEQ ID NO:634).

[0260] In some embodiments, the antibody comprises a light chain CDR2 sequence having the consensus sequence xSxxxS (SEQ ID NO:253). In some embodiments, the light chain CDR2 consensus sequence comprises the sequence [V/T/A]S[K/Y/Q][L/R][D/A/Y]S (SEQ ID NO:635).

[0261] In some embodiments, the antibody comprises a light chain CDR3 sequence having the consensus sequence xQxxxxPxT (SEQ ID NO:255). In some embodiments, the light chain CDR3 consensus sequence comprises the sequence

[W/Q/M]Q[GAV/Y/H][T/S/N/L][H/S/E][F/N/Y]P[Q/P/L/R/F]T (SEQ ID NO:636).

[0262] In some embodiments, the antibody comprises a heavy chain CDRl sequence having the consensus sequence xYxITxxxYWN (SEQ ID NO:317). In some embodiments, the heavy chain CDRl consensus sequence comprises the sequence [G/D]Y[S/P]IT[S/N/R][G/F/N][Y/F]YWN (SEQ ID NO:637).

[0263] In some embodiments, the antibody comprises a heavy chain CDRl sequence having the consensus sequence GxTFTDYYxN (SEQ ID NO:318). In some embodiments, the heavy chain CDRl consensus sequence comprises the sequence G[H/Y/F/N]TFTDYY[M/I]N (SEQ ID NO:638).

[0264] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: 1 comprises a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:241, 242, 317, and 318. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:64-78.

[0265] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: 1 comprises a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:245 and 246. In some embodiments, the anti-alpha- synuclein antibody comprises a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:83-115.

[0266] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: 1 comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:249. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 122-141.

[0267] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: l comprises a light chain CDRl comprising the amino acid sequence of SEQ ID NO:251. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:206-220.

[0268] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: l comprises a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:253. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:224-228.

[0269] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: l comprises a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:255. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232-237.

[0270] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: l comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:241, 242, 317, and 318; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:245-246; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:249; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:251; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:253; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:255. [0271] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 71-105 of SEQ ID NO: l comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:64-78; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:83-l 15; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 122-141 ; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206-220; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:224-228; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232-237.

2D01

[0272] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:64, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:83, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 122. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 83, 122, 218, 228, and 237, respectively.

[0273] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:4. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4.

[0274] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 149. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 149. [0275] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:4 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 149. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 149.

[0276] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 83, 122, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 149).

2E01

[0277] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:64, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:83, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 122. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:206, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:224, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:232. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 83, 122, 207, 224, and 232, respectively.

[0278] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 5. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 5.

[0279] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 150. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150.

[0280] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:5 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 150. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:5 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150.

[0281] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 83, 122, 206, 224, and 232, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:5 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150).

2F01

[0282] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:64, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:84, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 123. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:207, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 84, 123, 207, 228, and 237, respectively.

[0283] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:6. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:6.

[0284] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 151. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 151.

[0285] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:6 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 151. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:6 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 151.

[0286] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 84, 123, 207, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:6 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 151). 1F02

[0287] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:83, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 124. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 83, 124, 218, 228, and 237, respectively.

[0288] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:7. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7.

[0289] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 152. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 152.

[0290] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:7 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 152. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 152.

[0291] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 83, 124, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 152).

2A02

[0292] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:85, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 125. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:208, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:236. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 85, 125, 208, 228, and 237, respectively.

[0293] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 8. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

[0294] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 153. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 153. [0295] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:8 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 153. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 153.

[0296] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 85, 125, 208, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 153).

1H02

[0297] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:71, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:86, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:71, 86, 126, 209, 228, and 237, respectively.

[0298] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:9. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:9.

[0299] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 154. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:4

[0300] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:9 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 154. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:9 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 154.

[0301] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:71, 86, 126, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:9 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 154).

2B02

[0302] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:87, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 87, 126, 218, 228, and 237, respectively.

[0303] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 10. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10.

[0304] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 155. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 155.

[0305] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 10 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 155. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 155.

[0306] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 87, 126, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 155). 2C02

[0307] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:72, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:86, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:207, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:72, 86, 126, 207, 228, and 237, respectively.

[0308] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 1 1. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 1.

[0309] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 156. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 156.

[0310] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 1 1 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 156. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 1 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 156.

[0311] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:75, 86, 126, 207, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 1 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 156).

2D02

[0312] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:88, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 126. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 88, 126, 218, 228, and 237, respectively.

[0313] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 12. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12.

[0314] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 157. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 157. [0315] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO: 12 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 157. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 157.

[0316] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 88, 126, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 157).

1A03

[0317] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:89, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 127. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 127, 218, 228, and 237, respectively.

[0318] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 13. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13.

[0319] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 158. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 158.

[0320] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO: 13 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 158. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 158.

[0321] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 127, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 158).

2A03

[0322] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:84, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 127. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:210, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 127, 210, 228, and 237, respectively.

[0323] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 14. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14.

[0324] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 159. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 159.

[0325] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 14 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 159. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 159.

[0326] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 127, 210, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 159). 2G02

[0327] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:64, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:83, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 127. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 83, 127, 220, 228, and 237, respectively.

[0328] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 15. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[0329] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 160. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 160.

[0330] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 15 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 160. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 160.

[0331] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 83, 127, 220, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 160).

1D01

[0332] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:66, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:90, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 128. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:66, 90, 128, 218, 228, and 237, respectively.

[0333] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 16. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 16.

[0334] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 161. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 161. [0335] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO: 16 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 161. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 16 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 161.

[0336] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:66, 90, 128, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 16 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 161).

2C03

[0337] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:66, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:91, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 128. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:66, 91, 128, 218, 228, and 237, respectively.

[0338] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 17. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17.

[0339] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 162. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 162.

[0340] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO: 17 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 162. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 162.

[0341] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:66, 91, 128, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 162).

1D02

[0342] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:67, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:92, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:21 1, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 92, 129, 21 1, 228, and 237, respectively.

[0343] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 18. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 18.

[0344] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 163. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 163.

[0345] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 18 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 163. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 18 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 163.

[0346] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 92, 129, 21 1, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 18 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 163). 2A04

[0347] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:67, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:93, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:212, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 93, 129, 212, 228, and 237, respectively.

[0348] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 19. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19.

[0349] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 164. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 164.

[0350] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 19 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 164. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 164.

[0351] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 93, 129, 212, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 164).

2B04

[0352] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:68, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:94, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:68, 94, 129, 209, 228, and 237, respectively.

[0353] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:20. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:20.

[0354] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 165. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 165. [0355] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:20 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 165. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:20 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 165.

[0356] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:68, 94, 129, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:20 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 165).

2C04

[0357] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:67, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:95, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 95, 129, 209, 228, and 237, respectively.

[0358] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:21. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21.

[0359] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 166. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 166.

[0360] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:21 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 166. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 166.

[0361] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 95, 129, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 166).

2E03

[0362] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:78, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:96, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:78, 96, 129, 218, 228, and 237, respectively.

[0363] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:22. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:22.

[0364] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 157. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 157.

[0365] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:22 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 157. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:22 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 157.

[0366] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:78, 96, 129, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:22 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 157). 2E04

[0367] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:68, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:97, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:68, 97, 129, 209, 228, and 237, respectively.

[0368] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:23. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23.

[0369] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 167. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 167.

[0370] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:23 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 167. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 167.

[0371] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:68, 97, 129, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 167).

2G04

[0372] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:67, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:98, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 129. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 98, 129, 209, 228, and 237, respectively.

[0373] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:24. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:24.

[0374] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 168. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168. [0375] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:24 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 168. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:24 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168.

[0376] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 98, 129, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:24 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168).

2H04

[0377] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:69, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 199, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 130. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:219, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:69, 99, 130, 219, 228, and 237, respectively.

[0378] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:25. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25.

[0379] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 169. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 169.

[0380] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:25 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 169. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 169.

[0381] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:69, 99, 130, 219, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 169).

2B05

[0382] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:70, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 100, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 131. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:70, 100, 131, 220, 228, and 237, respectively.

[0383] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:26. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:26.

[0384] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 170. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 170.

[0385] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:26 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 170. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:26 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 170.

[0386] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:70, 100, 131, 220, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:26 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 170). 2C05

[0387] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:84, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 131. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 131, 209, 228, and 237, respectively.

[0388] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:27. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27.

[0389] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 171. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 171.

[0390] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:27 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 171. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 171.

[0391] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 131, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 171).

2D05

[0392] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:71, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:89, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:213, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:225, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:234. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:71, 89, 132, 213, 225, and 234, respectively.

[0393] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:28. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:28.

[0394] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 172. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 172. [0395] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:28 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 172. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:28 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 172.

[0396] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:71, 89, 132, 213, 225, and 234, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:28 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 172).

2E05

[0397] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:71, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:89, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:71, 89, 132, 209, 228, and 237, respectively.

[0398] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:29. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:29.

[0399] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 173. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 173.

[0400] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:29 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 173. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:29 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 173.

[0401] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:71, 89, 132, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:29 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 173).

2G05

[0402] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:64, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:84, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 133. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 84, 133, 209, 228, and 237, respectively.

[0403] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:30. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:30.

[0404] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 174. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 174.

[0405] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:30 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 174. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:30 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 174.

[0406] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:64, 84, 133, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:30 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 174). 2A06

[0407] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 101, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 134. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 101, 134, 218, 228, and 237, respectively.

[0408] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 31. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 31.

[0409] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 175. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 175.

[0410] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:31 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 175. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 175.

[0411] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 101, 134, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 175).

2H05

[0412] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:73, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 102, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 134. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:235. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:73, 102, 134, 209, 228, and 235, respectively.

[0413] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:32. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:32.

[0414] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 176. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 176. [0415] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:32 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 176. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:32 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 176.

[0416] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:73, 102, 134, 209, 228, and 235, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 181).

2C06

[0417] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:74, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 103, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 135. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:74, 103, 135, 218, 228, and 237, respectively.

[0418] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:33. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33.

[0419] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 177. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 177.

[0420] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:33 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 177. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 177.

[0421] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:74, 103, 135, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 177).

2G06

[0422] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:89, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 136. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 136, 218, 228, and 237, respectively.

[0423] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:34. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34.

[0424] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 178. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 178.

[0425] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:34 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 178. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 178.

[0426] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 136, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 178). 2H06

[0427] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:75, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 104, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 136. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:212, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:75, 104, 136, 212, 228, and 237, respectively.

[0428] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:35. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35.

[0429] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 179. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 179.

[0430] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:35 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 179. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 179.

[0431] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:75, 104, 136, 212, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 179).

1D03

[0432] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:73, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 101, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 137. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:73, 101, 137, 220, 228, and 237, respectively.

[0433] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:36. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:36.

[0434] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 180. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 180. [0435] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:36 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 180. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:36 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 180.

[0436] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:73, 101, 137, 220, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:36 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 180).

2A07

[0437] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 101, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 137. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:214, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 101, 137, 214, 228, and 237, respectively.

[0438] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:37. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37.

[0439] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 181. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 181.

[0440] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:37 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 181. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 181.

[0441] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 101, 137, 214, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 181).

2B07

[0442] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 105, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 137. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:206, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:224, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:233. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 105, 137, 206, 224, and 233, respectively.

[0443] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:38. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:38.

[0444] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 182. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 182.

[0445] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:38 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 182. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:38 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 182.

[0446] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 105, 137, 206, 224, and 233, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:38 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 182). 1E03

[0447] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 106, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 185, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 106, 138, 218, 228, and 237, respectively.

[0448] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:39. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39.

[0449] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 183. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 183.

[0450] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:39 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 183. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 183.

[0451] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 106, 138, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 183).

1F03

[0452] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:76, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 107, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:76, 107, 138, 209, 228, and 237, respectively.

[0453] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:40. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:40.

[0454] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 184. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184. [0455] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:40 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 184. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:40 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184.

[0456] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:76, 107, 138, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:40 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184).

1G03

[0457] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 108, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 108, 138, 209, 228, and 237, respectively.

[0458] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:41. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:41.

[0459] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 185. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 185.

[0460] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:41 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 185. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:41 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 185.

[0461] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 108, 138, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:41 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 185).

2A08

[0462] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 106, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 106, 138, 209, 228, and 237, respectively.

[0463] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:42. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:42.

[0464] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 184. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184.

[0465] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:42 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 184. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:42 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184.

[0466] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 106, 138, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:42 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184). 2B08

[0467] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 106, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 106, 138, 220, 228, and 237, respectively.

[0468] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:43. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:43.

[0469] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 186. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186.

[0470] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:43 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 186. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:43 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186.

[0471] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 106, 138, 220, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:43 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186).

2C07

[0472] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 109, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 109, 138, 218, 228, and 237, respectively.

[0473] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:44. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:44.

[0474] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 187. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 187. [0475] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:44 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 187. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:44 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 187.

[0476] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 109, 138, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:44 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 187).

2D07

[0477] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 10, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:212, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 10, 138, 212, 228, and 237, respectively.

[0478] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:45. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:45.

[0479] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 188. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 188.

[0480] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:45 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 188. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:45 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 188.

[0481] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 10, 138, 212, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:45 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 188).

2G07

[0482] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:84, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 138, 209, 228, and 237, respectively.

[0483] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:46. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:46.

[0484] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 184. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184.

[0485] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:46 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 184. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:46 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184.

[0486] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 138, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:46 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 184). 2H07

[0487] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:84, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:215, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 138, 215, 228, and 237, respectively.

[0488] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:47. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:47.

[0489] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 189. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 189.

[0490] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:47 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 189. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:47 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 189.

[0491] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 84, 138, 215, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:47 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 189).

2D08

[0492] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:89, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 139. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 139, 218, 228, and 237, respectively.

[0493] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:48. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:48.

[0494] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 190. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 190. [0495] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:48 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 190. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:48 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 190.

[0496] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 139, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:48 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 190).

2B09

[0497] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 1 1, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:212, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:226, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 1 1, 140, 212, 226, and 237, respectively.

[0498] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:49. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:49.

[0499] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 191. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 191.

[0500] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:49 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 191. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:49 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 191.

[0501] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 1 1, 140, 212, 226, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:49 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 190).

2C09

[0502] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:77, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 12, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:77, 1 12, 140, 218, 228, and 237, respectively.

[0503] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:50. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:50.

[0504] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 192. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 192.

[0505] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:50 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 192. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:50 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 192.

[0506] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:77, 1 12, 140, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 50 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 192). 2D09

[0507] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 13, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 13, 140, 209, 228, and 237, respectively.

[0508] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 51. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:51.

[0509] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 193. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 193.

[0510] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:51 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 193. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:51 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 193.

[0511] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 13, 140, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:51 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 193).

2F08

[0512] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 14, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:216, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:227, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:236. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 14, 140, 216, 227, and 236, respectively.

[0513] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:52. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:52.

[0514] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 194. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 194. [0515] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:52 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 194. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:52 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 194.

[0516] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 14, 140, 216, 227, and 236, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 52 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 194).

2G08

[0517] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:89, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 140, 209, 228, and 237, respectively.

[0518] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:53. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:53.

[0519] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 195. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 195.

[0520] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%>, or at least 99% sequence identity) to SEQ ID NO:53 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 195. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:53 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 195.

[0521] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 89, 140, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 53 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 195).

2H08

[0522] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:87, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:209, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 87, 140, 209, 228, and 237, respectively.

[0523] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:54. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:54.

[0524] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 196. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 196.

[0525] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:54 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 196. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:54 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 196.

[0526] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 87, 140, 209, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 196). 2E09

[0527] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:67, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 15, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 141. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:217, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 1 15, 141, 217, 228, and 237, respectively.

[0528] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 55. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 55.

[0529] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 197. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 197.

[0530] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:55 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 197. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:55 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 197.

[0531] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:67, 1 15, 141, 217, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:55 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 197).

Anti-alpha-synuclein antibodies that recognize an epitope within residues 66-85

[0532] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 66-85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 66-80 of SEQ ID NO: l or recognizes an epitope within residues 71-85 of SEQ ID NO: 1. In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0533] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues V66, G67, G68, A69, V70, V71, T72, G73, V74, T75, A76, V77, A78, Q79, K80, T81, V82, E83, G84, or A85 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, or all 20 of the following residues V66, G67, G68, A69, V70, V71, T72, G73, V74, T75, A76, V77, A78, Q79, K80, T81, V82, E83, G84, or A85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to non-contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: 1.

[0534] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V66 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G67 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G68 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A69 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue V70 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V71 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T72 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G73 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V74 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T75 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A76 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V77 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A78 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q79 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K80 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T81 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V82 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E83 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G84 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A85 of SEQ ID NO: 1.

[0535] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence VGGAVVTGVTAVAQK (SEQ ID NO:260), which corresponds to residues 66-80 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:260 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0536] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence VTGVTAVAQKTVEGA (SEQ ID NO:261), which corresponds to residues 71-85 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:261 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0537] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 79 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:79; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 116 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 116; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 142 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 142; (d) a light chain CDRl having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:221; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:229 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:229; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:238 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:238.

[0538] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDRl of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDRl of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0539] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:79; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:221; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

[0540] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

2F09

[0541] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:79, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 16, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 142. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:221, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:229, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:238. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:79, 1 16, 142, 221, 229, and 238, respectively.

[0542] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:56. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:56.

[0543] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 198. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 198.

[0544] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:56 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 198. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:56 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 198.

[0545] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:79, 1 16, 142, 221, 229, and 238, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 198).

Anti-alpha-synuclein antibodies that recognize an epitope within residues 81-1 15

[0546] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 81-1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 81-95 of SEQ ID NO: l, recognizes an epitope within residues 86-100 of SEQ ID NO: l, recognizes an epitope within residues 91-105 of SEQ ID NO: l, or recognizes an epitope within residues 101-1 15 of SEQ ID NO: l . In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0547] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues T81, V82, E83, G84, A85, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, E105, G106, A107, P108, Q109, E1 10, Gi l l, 11 12, L1 13, E1 14, or D1 15 of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody binds to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or all 35 of the following residues T81, V82, E83, G84, A85, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, E105, G106, A107, P108, Q109, El 10, Gi l l, 11 12, LI 13, El 14, or D1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to non-contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: l . [0548] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T81 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V82 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E83 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G84 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G86 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue S87 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue 188 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A89 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A90 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A91 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T92 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue G93 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue F94 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V95 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K96 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K97 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D98 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q99 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue LI 00 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue G101 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K102 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N103 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 04 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G106 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A107 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 08 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue Q 109 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 10 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Gl 1 1 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue II 12 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue LI 13 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 14 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Dl 15 of SEQ ID NO: 1.

[0549] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence TVEGAGSIAAATGFV (SEQ ID NO:262), which corresponds to residues 81-95 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:262 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0550] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GSIAAATGFVKKDQL (SEQ ID NO:258), which corresponds to residues 86-100 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:258 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0551] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence ATGFVKKDQLGKNEE (SEQ ID NO:259), which corresponds to residues 91-105 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:259 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0552] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GKNEEGAPQEGILED (SEQ ID NO:265), which corresponds to residues 101-1 15 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:265 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0553] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1 17, 1 18, and 247 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 1 17, 1 18, and 247; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:237 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:237.

[0554] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0555] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 1 17, 1 18, and 247; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0556] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0557] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:57-58. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 199-201. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 57-58, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs: 199- 201.

[0558] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:57-58. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs: 199-201. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:57-58, and a light chain variable region comprising the amino acid of any one of SEQ ID NOs: 199- 201.

[0559] In some embodiments, an anti-alpha-synuclein antibody comprises one or more sequences that are variants of one or more consensus sequences or are encompassed by one or more consensus sequences. In some embodiments, one or more consensus sequences can be identified for antibodies that recognize the same or similar epitope as 2F10. Exemplary 2F 10- like consensus sequences include SEQ ID NOs:243, 247, 250, 252, 254, 319, and 320. In the consensus sequences of SEQ ID NOs:243, 247, 250, 252, 254, 319, and 320, the capitalized letter represents an amino acid residue that is absolutely conserved among the aligned sequences (e.g., aligned CDR sequences), while "x" represents an amino acid residue that is not absolutely conserved among the aligned sequences. While "x" can be any amino acid, it will be appreciated that when selecting an amino acid to insert at a position marked by an "x" that in some embodiments, the amino acid is selected from those amino acids found at the corresponding position in the aligned sequences.

[0560] In some embodiments, the antibody comprises a heavy chain CDR1 sequence having the consensus sequence xYSITSGYY (SEQ ID NO:243). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence [D/G]YSITSGYY (SEQ ID NO:639).

[0561] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence ISDDGRNxYNPSLKN (SEQ ID NO:247). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence ISDDGRN[D/N]YNPSLKN (SEQ ID NO:640).

[0562] In some embodiments, the antibody comprises a heavy chain CDR3 sequence having the consensus sequence xRGDxGP (SEQ ID NO:250). In some embodiments, the heavy chain CDR3 consensus sequence comprises the sequence [L/V]RGD[N/Y]GP (SEQ ID NO:641).

[0563] In some embodiments, the antibody comprises a light chain CDR1 sequence having the consensus sequence KSxQSLxDxDGxTYLN (SEQ ID NO:252). In some embodiments, the light chain CDR1 consensus sequence comprises the sequence KS[S/N]QSL[L/F]D[S/Y]DG[E/K]TYLN (SEQ ID NO:642). [0564] In some embodiments, the antibody comprises a light chain CDR2 sequence having the consensus sequence VSKxDS (SEQ ID NO:254). In some embodiments, the light chain CDR2 consensus sequence comprises the sequence VSK[L/Q]DS (SEQ ID NO:643).

[0565] In some embodiments, the antibody comprises a heavy chain CDR1 sequence having the consensus sequence xYSITSGYYWN (SEQ ID NO:319). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence [D/G]YSITSGYYWN (SEQ ID NO:644).

[0566] In some embodiments, the antibody comprises a light chain CDR2 sequence having the consensus sequence LVSKxDS (SEQ ID NO:320). In some embodiments, the light chain CDR2 consensus sequence comprises the sequence LVSK[L/Q]DS (SEQ ID NO:645).

[0567] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: 1 comprises a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:243 and 319. In some embodiments, the anti-alpha- synuclein antibody comprises a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65 and 80.

[0568] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: 1 comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:247. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 117 and 118.

[0569] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: 1 comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:250. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 143 and 144.

[0570] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: 1 comprises a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:252. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:218, 219, and 222. [0571] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: 1 comprises a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:254 and 320. In some embodiments, the anti-alpha- synuclein antibody comprises a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228 and 230.

[0572] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: l comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:243 and 319; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:247; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:250; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:252; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:254 and 320; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

[0573] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 81-115 of SEQ ID NO: l comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65 and 80; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 117 and 118; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 143 and 144; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:218, 219, and 222; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228 and 230; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

1E04

[0574] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:80, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 117, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 143. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:80, 1 17, 143, 218, 228, and 237, respectively.

[0575] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:57. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:57.

[0576] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 199. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 199.

[0577] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:57 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 199. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:57 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 199.

[0578] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:80, 1 17, 143, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 57 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:209). 2G09

[0579] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:80, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 17, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 143. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:222, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:80, 1 17, 143, 222, 228, and 237, respectively.

[0580] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:57. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:57.

[0581] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:200. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:200.

[0582] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:57 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:200. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:57 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:200.

[0583] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:80, 1 17, 143, 222, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 57 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:200).

2F 10

[0584] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 18, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 144. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:219, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:230, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 18, 144, 219, 230, and 237, respectively.

[0585] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO: 58. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 58.

[0586] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:201. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:201. [0587] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:58 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:201. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:58 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:201.

[0588] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 1 18, 144, 219, 230, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:58 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:201).

Anti-alpha-synuclein antibodies that recognize an epitope within residues 36-50 and residues 86-1 15

[0589] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 36-50 of SEQ ID NO: 1 and recognizes an epitope within residues 86-1 15 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 36-50 of SEQ ID NO: l, recognizes an epitope within residues 86-100 of SEQ ID NO: 1, recognizes an epitope within residues 96-1 10 of SEQ ID NO: 1, or recognizes an epitope within residues 101-1 15 of SEQ ID NO: l . In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0590] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues G36, V37, L38, Y39, V40, G41, S42, K43, T44, K45, E46, G47, V48, V49, H50, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, L100, G101, K102, N103, E104, E105, G106, A107, P108, Q109, E1 10, Gi l l, 11 12, L1 13, El 14, or D1 15 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or all 45 of the following residues G36, V37, L38, Y39, V40, G41, S42, K43, T44, K45, E46, G47, V48, V49, H50, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, E105, G106, A107, P108, Q109, E110, Gi l l, 1112, L113, E114, or D115 of SEQ ID NCv l. In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to non-contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: l .

[0591] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G36 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V37 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue L38 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y39 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue V40 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G41 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue S42 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K43 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T44 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K45 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E46 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G47 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue V48 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V49 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue H50 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G86 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue S87 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue 188 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A89 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A90 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A91 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T92 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G93 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue F94 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K96 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K97 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D98 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue Q99 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue L100 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G101 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K 102 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N103 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E104 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 05 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G106 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A107 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue P108 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q 109 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 10 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Gi l l of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue 1112 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue LI 13 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 14 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue D115 of SEQ ID NO: l .

[0592] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GVLYVGSKTKEGVVH (SEQ ID NO:263), which corresponds to residues 36-50 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:263 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0593] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GSIAAATGFVKKDQL (SEQ ID NO:258), which corresponds to residues 86-100 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:258 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0594] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence KKDQLGKNEEGAPQE (SEQ ID NO:264), which corresponds to residues 96-1 10 of SEQ ID NO: 1. In some embodiments, the peptide sequence set forth in SEQ ID NO:264 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0595] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GKNEEGAPQEGILED (SEQ ID NO:265), which corresponds to residues 100-1 15 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:265 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0596] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 63 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:63; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1 19, 120, and 248 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 1 19, 120, and 248; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 145 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 145; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:220; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:228 or 316 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:228 or 316; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:239 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:239.

[0597] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of

(d) , the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0598] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:63; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 1 19, 120, and 248; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:220;

(e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228 or 316; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

[0599] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0600] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:59-61. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:202-204. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:59-61, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:202- 204.

[0601] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:59-61. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:202-204. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:59-61, and a light chain variable region comprising the amino acid of any one of SEQ ID NOs:202- 204.

[0602] In some embodiments, an anti-alpha-synuclein antibody comprises one or more sequences that are variants of one or more consensus sequences or are encompassed by one or more consensus sequences. In some embodiments, one or more consensus sequences can be identified for antibodies that recognize the same or similar epitope as 3C02. An exemplary 3C02-like consensus sequence includes SEQ ID NO:248. In the consensus sequence of SEQ ID NO:248, the capitalized letter represents an amino acid residue that is absolutely conserved among the aligned sequences (e.g., aligned CDR sequences), while "x" represents an amino acid residue that is not absolutely conserved among the aligned sequences. While "x" can be any amino acid, it will be appreciated that when selecting an amino acid to insert at a position marked by an "x" that in some embodiments, the amino acid is selected from those amino acids found at the corresponding position in the aligned sequences.

[0603] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence I SRGS S AI Y YAD A VxG (SEQ ID NO:248). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence ISRGSSAIYYADAV[K/R]G (SEQ ID NO:646).

[0604] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 36-50 and 86-115 of SEQ ID NO: 1 comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:248. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 119 and 120.

[0605] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 36-50 and 86-115 of SEQ ID NO: 1 comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:63; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:248; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:220; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228 and 316; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

[0606] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 36-50 and 86-115 of SEQ ID NO: 1 comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:63; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 119 and 120; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145; (d) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:220; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228 and 316; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

1G06

[0607] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl sequence comprising the amino acid sequence of SEQ ID NO:63, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 119, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 145. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDRl sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:316, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:239. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 1 19, 145, 220, 316, and 239, respectively.

[0608] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:59. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:59.

[0609] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:202. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:202.

[0610] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:59 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:202. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:59 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:202.

[0611] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 1 19, 145, 220, 316, and 239, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 59 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:202). 3C02

[0612] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:63, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 1 19, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 145. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:239. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 1 19, 145, 220, 228, and 239, respectively.

[0613] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:60. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:60.

[0614] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:203. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:203.

[0615] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:60 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:203. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:60 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:203.

[0616] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 1 19, 145, 220, 228, and 239, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:60 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:203).

3D02

[0617] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:63, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 120, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 145. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:316, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:239. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 120, 145, 220, 316, and 239, respectively.

[0618] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:61. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:61.

[0619] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:204. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:204. [0620] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:61 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:204. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:61 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:204.

[0621] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:63, 120, 145, 220, 316, and 239, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:61 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:204).

Anti-alpha-synuclein antibodies that recognize an epitope within residues 86-105 or residues 121-140

[0622] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 86-105 of SEQ ID NO: 1 or recognizes an epitope within residues 121-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 86-100 of SEQ ID NO: l, recognizes an epitope within residues 91-105 of SEQ ID NO: l, recognizes an epitope within residues 121-135 of SEQ ID NO: l, or recognizes an epitope within residues 126-140 of SEQ ID NO: 1. In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0623] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, E105, D121, N122, E123, A124, Y125, E126, M127, P128, S 129, E130, E131, G132, Y133, Q134, D135, Y136, E137, P138, E139, or A140 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least 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, 32, 33, 34, 35, 36, 37, 38, 39, or all 40 of the following residues G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, L100, G101, K102, N103, E104, E105, D121, N122, E123, A124, Y125, E126, M127, P128, S129, E130, E131, G132, Y133, Q134, D135, Y136, E137, P138, E139, or A140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to non-contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: l .

[0624] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V71 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T72 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G73 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V74 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue T75 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A76 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V77 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A78 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q79 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K80 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T81 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V82 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue E83 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G84 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A85 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G86 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue S87 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue 188 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A89 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A90 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A91 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T92 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G93 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue F94 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V95 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K96 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K97 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D98 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue Q99 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue L100 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G101 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K 102 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N103 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E104 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 05 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D 121 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue N122 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E123 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A124 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E126 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Ml 27 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 28 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue SI 29 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue E130 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E131 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G132 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y133 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q134 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D135 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y136 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 37 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue PI 38 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E139 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A140 of SEQ ID NO: 1.

[0625] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence GSIAAATGFVKKDQL (SEQ ID NO:258), which corresponds to residues 86-100 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:258 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0626] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence ATGFVKKDQLGKNEE (SEQ ID NO:259), which corresponds to residues 91-105 of SEQ ID NO: l . In some embodiments, the peptide sequence set forth in SEQ ID NO:259 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0627] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence DNEAYEMPSEEGYQD (SEQ ID NO:323), which corresponds to residues 121-135 of SEQ ID NO: 1. In some embodiments, the peptide sequence set forth in SEQ ID NO:323 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0628] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence EMP SEEG YQD YEPE A (SEQ ID NO:322), which corresponds to residues 126-140 of SEQ ID NO: 1. In some embodiments, the peptide sequence set forth in SEQ ID NO:322 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below). [0629] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309- 310 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 311-312 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 31 1-312; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 206, 218, 301-304, and 313; (e) a light chain CDR2 having at least 90%) sequence identity to the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314; and (f) a light chain CDR3 having at least 90%) sequence identity to the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315.

[0630] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0631] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 311-312; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291 ; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315.

[0632] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0633] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:269-272 and 274-275. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:292-295 and 298-300. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:269-272 and 274-275, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:292-295 and 298-300.

[0634] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:269-272 and 274-275. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:292-295 and 298-300. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:269-272 274-275, and a light chain variable region comprising the amino acid of any one of SEQ ID NOs:292-295 and 298-300. Anti-alpha-synuclein antibodies that recognize an epitope within residues 86-105

[0635] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 86-105 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 276, and 277 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 276, and 277; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:279, 281, and 282 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:279, 281, and 282; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:285-287 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:285- 287; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:218, 301, and 302 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:218, 301, and 302; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:223 and 228 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:223 and 228; and (f) a light chain CDR3 having at least 90%) sequence identity to the amino acid sequence of any one of SEQ ID NOs:237 and 306 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:237 and 306.

[0636] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0637] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65, 276, and 277; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:279, 281, and 282; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:285-287; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:218, 301, and 302; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:223 and 228; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID SEQ ID NOs:237 and 306.

[0638] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0639] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:269-271. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:292-294. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:269-271, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:292- 294.

[0640] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:269-271. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:292-294. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:269-271, and a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:292-294.

Anti-alpha-synuclein antibodies that recognize an epitope within residues 86-105 and 121- 140

[0641] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 86-105 and 121-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:277 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:283 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:283; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:288 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:288; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 303 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:303; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:305 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 305; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:307 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NO:307.

[0642] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0643] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:283; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:288; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:303; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:305; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:307.

[0644] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0645] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:272. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:295. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:272, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:295.

[0646] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:272. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:295. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:272, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:295.

Anti-alpha-synuclein antibodies that recognize an epitope within residues 121-140 [0647] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 121-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:277 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:283 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:283; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:291 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:291; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 304 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:304; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:308 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NO: 308.

[0648] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0649] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:283; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:291; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:304; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 308. [0650] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0651] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:275. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:299 or 300. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:275, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:299 or 300.

[0652] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:299 or 300. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:299 or 300.

Anti-alpha-synuclein antibodies that recognize an epitope within residues 126-140

[0653] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 126-140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:278 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:278; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:284 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:284; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:290 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:290; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:206 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:206; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:224; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:232 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NO: 232.

[0654] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0655] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:278; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:284; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:290; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:206; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:224; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:232.

[0656] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). [0657] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:274. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:298. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:274, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:298.

[0658] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:274. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:298. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:274, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:298.

[0659] In some embodiments, an anti-alpha-synuclein antibody comprises one or more sequences that are variants of one or more consensus sequences or are encompassed by one or more consensus sequences. In the consensus sequences described below, the capitalized letter represents an amino acid residue that is absolutely conserved among the aligned sequences (e.g., aligned CDR sequences), while "x" represents an amino acid residue that is not absolutely conserved among the aligned sequences. While "x" can be any amino acid, it will be appreciated that when selecting an amino acid to insert at a position marked by an "x" that in some embodiments, the amino acid is selected from those amino acids found at the corresponding position in the aligned sequences.

[0660] In some embodiments, the antibody comprises a heavy chain CDR1 sequence having the consensus sequence GYSITSGxYWN (SEQ ID NO:309). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence GYSITSG[H/Y]YWN (SEQ ID NO:647).

[0661] In some embodiments, the antibody comprises a heavy chain CDR1 sequence having the consensus sequence GFxFNTYAMx (SEQ ID NO:310). In some embodiments, the heavy chain CDR1 consensus sequence comprises the sequence GF[T/S]FNTYAM[H/N] (SEQ ID NO:648).

[0662] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence RIRxKxxNYAT Y YAD S VDK (SEQ ID NO:311). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence RIR[S/T]K[S/I][N/S]NYATYYADSVDK (SEQ ID NO:649).

[0663] In some embodiments, the antibody comprises a heavy chain CDR2 sequence having the consensus sequence xxxxDGxxNYNPSLKN (SEQ ID NO:312). In some embodiments, the heavy chain CDR2 consensus sequence comprises the sequence [F/Y][I/L][S/T][D/Y]DG[I/N][N/K]NYNPSLKN (SEQ ID NO:650).

[0664] In some embodiments, the antibody comprises a light chain CDR1 sequence having the consensus sequence xSxxxxxxxxGxTYLx (SEQ ID NO:313). In some embodiments, the light chain CDR1 consensus sequence comprises the sequence [K/R]S[S/G][Q/R][S/D][I/L][I/V/L][D/H][S/V][D/Y/N]G[E/S/N]TYL[N/E] (SEQ ID NO:651).

[0665] In some embodiments, the antibody comprises a light chain CDR2 sequence having the consensus sequence xxSxxxS (SEQ ID NO:314). In some embodiments, the light chain CDR2 consensus sequence comprises the sequence [K/D/L][V/T]S[N/K/T][R/L][F/A/D]S (SEQ ID NO:652).

[0666] In some embodiments, the antibody comprises a light chain CDR3 sequence having the consensus sequence xQxxxxPxT (SEQ ID NO:315). In some embodiments, the light chain CDR3 consensus sequence comprises the sequence

[QAV/F]Q[W/G][S/T][S/H/R/L][N/F/V/G/I]P[P/Q/L]T (SEQ ID NO:653).

[0667] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: 1 comprises a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:309 and 310. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65 and 276-278.

[0668] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: 1 comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:312. In some embodiments, the anti-alpha-synuclein antibody comprises a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:279-283.

[0669] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: 1 comprises a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:313. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:285-291.

[0670] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: 1 comprises a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:314. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, and 305.

[0671] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: 1 comprises a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:315. In some embodiments, the anti-alpha-synuclein antibody comprises a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232, 236, 306-308.

[0672] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: l comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:309 and 310; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:312; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:285-291; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:313; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:314; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:315. [0673] In some embodiments, an anti-alpha-synuclein antibody that recognizes an epitope within residues 86-105 or 121-140 of SEQ ID NO: l comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65 and 276-278; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:279-283; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:285-291 ; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206, 218, and 301-204; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, and 305; and (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232, 236, and 306-308.

4F5

[0674] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:276, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:279, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:285. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:276, 279, 285, 218, 228, and 237, respectively.

[0675] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:269. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:269.

[0676] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:292. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:292.

[0677] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:269 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:292. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:269 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:292.

[0678] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:276, 279, 285, 218, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:269 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:292).

1G4

[0679] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:277, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:281, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:286. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:301, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:306. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 281, 286, 301, 223, and 306, respectively.

[0680] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:270. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:270.

[0681] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:293. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:293.

[0682] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:270 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:293. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:270 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:293.

[0683] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 281, 286, 301, 223, and 306, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:270 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:293).

16D4

[0684] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:282, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:287. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:302, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:237. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 282, 287, 302, 228, and 237, respectively.

[0685] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:271. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:271.

[0686] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:294. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:294.

[0687] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:271 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:294. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:271 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:294.

[0688] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:65, 282, 287, 302, 228, and 237, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:271 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:294). 28B6

[0689] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:277, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:283, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:288. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:303, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:305, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:307. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 283, 288, 303, 305, and 307, respectively.

[0690] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:272. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:272.

[0691] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:295. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:295.

[0692] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:272 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:295. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:272 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:295.

[0693] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 283, 288, 303, 305, and 307, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:272 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:295).

46D2

[0694] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:278, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:284, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:290. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:206, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:224, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:232. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:278, 284, 290, 206, 224, and 232, respectively.

[0695] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:274. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:274.

[0696] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:298. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:298. [0697] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:274 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:298. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:274 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:298.

[0698] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:278, 284, 290, 206, 224, and 232, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:274 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:298).

1F4

[0699] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:277, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:283, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:291. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:304, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 308. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 283, 291, 304, 223, and 308, respectively.

[0700] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:275. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275.

[0701] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:299. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:299.

[0702] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:275 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:299. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:299.

[0703] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 283, 291, 304, 223, and 308, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:299).

1F4 (C^S Variant)

[0704] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:277, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:283, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:291. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:304, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 308. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 283, 291, 304, 223, and 308, respectively.

[0705] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:275. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275.

[0706] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:300. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:300.

[0707] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:275 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:300. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:300.

[0708] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 283, 291, 304, 223, and 308, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:275 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 300). Anti-alpha-synuclein antibodies that recognize an epitope within residues 61-140

[0709] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 61-140 of SEQ ID NO: l . In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0710] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues E61, Q62, V63, T64, N65, V66, G67, G68, A69, V70, V71, T72, G73, V74, T75, A76, V77, A78, Q79, K80, T81, V82, E83, G84, A85, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, E105, G106, A107, P108, Q109, E1 10, Gi l l, 11 12, L1 13, E1 14, D1 15, M1 16, P1 17, V1 18, D1 19, P120, D121, N122, E123, A124, Y125, E126, M127, P128, S 129, E130, E131, G132, Y133, Q134, D135, Y136, E137, P138, E139, or A140 of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody binds to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 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, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or all 80 of the following residues E61, Q62, V63, T64, N65, V66, G67, G68, A69, V70, V71, T72, G73, V74, T75, A76, V77, A78, Q79, K80, T81, V82, E83, G84, A85, G86, S87, 188, A89, A90, A91, T92, G93, F94, V95, K96, K97, D98, Q99, LlOO, GlOl, K102, N103, E104, E105, G106, A107, P108, Q109, El 10, Gi l l, 11 12, L1 13, E1 14, D1 15, M1 16, P1 17, V1 18, D1 19, P120, D121, N122, E123, A124, Y125, E126, M127, P128, S 129, E130, E131, G132, Y133, Q134, D135, Y136, E137, P138, E139, or A140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to non-contiguous residues of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: l .

[0711] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E61 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q62 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V63 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T64 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue N65 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V66 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G67 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G68 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A69 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V70 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V71 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T72 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue G73 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V74 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T75 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A76 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V77 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A78 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q79 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K80 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue T81 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V82 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E83 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G84 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A85 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G86 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue S87 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue 188 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue A89 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A90 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A91 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue T92 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G93 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue F94 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue V95 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K96 of SEQ ID NO: l . In some embodiments, an anti-alpha- synuclein antibody binds to at least residue K97 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D98 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q99 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue LI 00 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G101 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue K 102 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N103 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 04 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue E105 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G106 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A107 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 08 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q109 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 10 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Gl 11 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue II 12 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue LI 13 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 14 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D 115 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Ml 16 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 17 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue VI 18 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Dl 19 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 20 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue D121 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue N122 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E123 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A124 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y125 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E126 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue M127 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue PI 28 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue SI 29 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E130 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E131 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G132 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y133 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q134 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D135 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y136 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue E137 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue P138 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E139 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A140 of SEQ ID NO: l .

[0712] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence

EQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEE GAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA (SEQ ID NO:321), which corresponds to residues 61-140 of SEQ ID NO: 1. In some embodiments, the peptide sequence set forth in SEQ ID NO:321 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below).

[0713] In some embodiments, an anti-alpha-synuclein antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:277 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:280 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:280; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:289 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:289; (d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:304 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:304; (e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:308 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:308.

[0714] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f). In some embodiments, a CDR having up to two amino acid substitutions has one amino acid substitution relative to the reference sequence. In some embodiments, a CDR having up to two amino acid substitutions has two amino acid substitutions relative to the reference sequence. In some embodiments, the up to two amino acid substitutions are conservative substitutions.

[0715] In some embodiments, the antibody comprises one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:277; (b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:280; (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:289; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:304; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 308.

[0716] In some embodiments, an anti-alpha-synuclein antibody comprises two, three, four, five, or all six of (a)-(f). In some embodiments, an anti-alpha-synuclein antibody comprises the heavy chain CDR1 of (a), the heavy chain CDR2 of (b), and the heavy chain CDR3 of (c). In some embodiments, an anti-alpha-synuclein antibody comprises the light chain CDR1 of (d), the light chain CDR2 of (e), and the light chain CDR3 of (f).

[0717] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:273. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:296-297. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:273, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to any one of SEQ ID NOs:296-297.

[0718] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273. In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of any one of SEQ ID NOs:296-297. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273, and a light chain variable region comprising the amino acid of any one of SEQ ID NOs:296-297.

3E12

[0719] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:277, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:280, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:289. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:304, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 308. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 280, 289, 304, 223, and 308, respectively.

[0720] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:273. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273.

[0721] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:296. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:296.

[0722] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:273 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:296. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:296.

[0723] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 280, 289, 304, 223, and 308, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:296).

3D5

[0724] In some embodiments, an anti-alpha-synuclein antibody recognizes an epitope within residues 126-140 of SEQ ID NO: 1. In some embodiments, the antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable regions as described herein (e.g., as described in Table 15 below).

[0725] In some embodiments, an anti-alpha-synuclein antibody binds to at least one of the following residues E126, M127, P128, S 129, E130, E131, G132, Y133, Q134, D135, Y136, E137, P138, E139, or A140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, or all 15 of the following residues E126, M127, P128, S 129, E130, E131, G132, Y133, Q134, D135, Y136, E137, P138, E139, or A140 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to contiguous residues of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to non-contiguous residues of SEQ ID NO: l . In some embodiments, an anti- alpha-synuclein antibody binds to both contiguous and non-contiguous residues of SEQ ID NO: l .

[0726] In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E126 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue M127 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue P128 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue SI 29 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue E130 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E131 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue G132 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y133 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Q134 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue D135 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue Y136 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue El 37 of SEQ ID NO: 1. In some embodiments, an anti-alpha- synuclein antibody binds to at least residue PI 38 of SEQ ID NO: 1. In some embodiments, an anti-alpha-synuclein antibody binds to at least residue E139 of SEQ ID NO: l . In some embodiments, an anti-alpha-synuclein antibody binds to at least residue A140 of SEQ ID NO: 1.

[0727] In some embodiments, an anti-alpha-synuclein antibody specifically binds to a peptide having the sequence EMP SEEG YQD YEPE A (SEQ ID NO:322), which corresponds to residues 126-140 of SEQ ID NO: 1. In some embodiments, the peptide sequence set forth in SEQ ID NO:322 is sufficient for binding to the anti-alpha-synuclein antibody. In some embodiments, the anti-alpha-synuclein antibody further comprises one or more complementarity determining region (CDR), heavy chain variable region, and/or light chain variable region sequences as described herein (e.g., as described in Table 15 below). [0728] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:277, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:280, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:289. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:304, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 308. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 280, 289, 304, 223, and 308, respectively.

[0729] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:273. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273.

[0730] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:297. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:297.

[0731] In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:273 and further comprises a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity (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%, or at least 99% sequence identity) to SEQ ID NO:297. In some embodiments, an anti-alpha-synuclein antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273 and further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:297. [0732] In some embodiments, an anti-alpha-synuclein antibody is an antibody that competes for binding with an antibody as described herein (e.g., an antibody comprising a heavy chain CDRl-3 and a light chain CDRl-3 comprising the amino acid sequences of SEQ ID NOs:277, 280, 289, 304, 223, and 308, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:273 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:297).

Preparation of antibodies

[0733] For preparing an anti-alpha-synuclein antibody, many techniques known in the art can be used. In some embodiments, antibodies are prepared by immunizing an animal or animals (e.g., mice, rabbits, or rats) with an antigen or a mixture of antigens for the induction of an antibody response. In some embodiments, the antigen or mixture of antigens is administered in conjugation with an adjuvant (e.g., Freund' s or Ribi adjuvant).

[0734] One embodiment therefore includes an antigen composition for immunizing an animal, comprising monomelic alpha-synuclein, oligomeric alpha-synuclein, and optionally an adjuvant, for example, Freund' s adjuvant. In one example, the antigen composition comprises a mixture of oligomeric alpha-synuclein protein in equilibrium with monomelic alpha- synuclein protein. In one example, the antigen composition is enriched for oligomeric alpha- synuclein protein compared to monomeric alpha-synuclein protein. In one example, the antigen composition comprises about 80% oligomeric and about 20% monomeric alpha- synuclein protein. Methods of separating or purifying oligomeric alpha-synuclein from monomeric alpha synuclein protein are known in the art, and include size exclusion chromatography. Another embodiment includes a method of producing an anti-alpha- synuclein antibody, comprising administering to an animal an antigen composition described herein, such as a mixture of monomeric alpha-synuclein, oligomeric alpha-synuclein, and optionally an adjuvant.

[0735] After an initial immunization, one or more subsequent booster injections of the antigen or antigens may be administered to improve antibody production. Following immunization, antigen-specific B cells are harvested, e.g., from the spleen and/or lymphoid tissue. For generating monoclonal antibodies, multiple methods may be used. In one embodiment, the B cells are fused with myeloma cells, which are subsequently screened for antigen specificity. Methods of preparing antibodies are also described in the Examples section below. [0736] The genes encoding the heavy and light chains of an antibody of interest can be cloned from a cell, e.g., the genes encoding a monoclonal antibody can be cloned from a hybridoma and used to produce a recombinant monoclonal antibody. Gene libraries encoding heavy and light chains of monoclonal antibodies can also be made from hybridomas or splenocytes or lymphocytes (e.g. B cells from an immunized animal). Alternatively, phage or yeast display technology can be used to identify antibodies and fragments that specifically bind to selected antigens via generation of immune Fab or scFv libraries and screening on recombinant antigen. Techniques for the production of single chain antibodies or recombinant antibodies can also be adapted to produce antibodies. Antibodies can also be made bispecific, i.e., able to recognize two different antigens. Antibodies can also be heteroconjugates, e.g., two covalently joined antibodies, or immunotoxins.

[0737] Antibodies can be produced using any number of expression systems, including prokaryotic and eukaryotic expression systems. In some embodiments, the expression system is a mammalian cell expression, such as a hybridoma, or a CHO cell expression system. Many such systems are widely available from commercial suppliers. In embodiments in which an antibody comprises both a VH and VL region, the VH and VL regions may be expressed using a single vector, e.g., in a di-cistronic expression unit, or under the control of different promoters. In other embodiments, the VH and VL region may be expressed using separate vectors. A VH or VL region as described herein may optionally comprise a methionine at the N-terminus.

[0738] In some embodiments, the antibody is a chimeric antibody. Methods for making chimeric antibodies are known in the art. For example, chimeric antibodies can be made in which the antigen binding region (heavy chain variable region and light chain variable region) from one species, such as a mouse, is fused to the effector region (constant domain) of another species, such as a human. As another example, "class switched" chimeric antibodies can be made in which the effector region of an antibody is substituted with an effector region of a different immunoglobulin class or subclass.

[0739] In some embodiments, the antibody is a humanized antibody. Generally, a non- human antibody is humanized in order to reduce its immunogenicity. Humanized antibodies typically comprise one or more variable regions (e.g., CDRs) or portions thereof that are non- human (e.g., derived from a mouse variable region sequence), and possibly some framework regions or portions thereof that are non-human, and further comprise one or more constant regions that are derived from human antibody sequences. Methods for humanizing non-human antibodies are known in the art. Transgenic mice, or other organisms such as other mammals, can be used to express humanized or human antibodies. Other methods of humanizing antibodies include, for example, variable region resurfacing, CDR grafting, grafting specificity-determining residues (SDR), guided selection, and framework shuffling.

[0740] As an alternative to humanization, fully human antibodies can be generated. As a non-limiting example, transgenic animals (e.g., mice) can be produced that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, it has been described that the homozygous deletion of the antibody heavy-chain joining region (JH) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge. As another example, human antibodies can be produced by hybridoma-based methods, such as by using primary human B cells for generating cell lines producing human monoclonal antibodies.

[0741] Human antibodies can also be produced using phage display or yeast display technology. In phage display, repertoires of variable heavy chain and variable light chain genes are amplified and expressed in phage display vectors. In some embodiments, the antibody library is a natural repertoire amplified from a human source. In some embodiments, the antibody library is a synthetic library made by cloning heavy chain and light chain sequences and recombining to generate a large pool of antibodies with different antigenic specificity. Phage typically display antibody fragments (e.g., Fab fragments or scFv fragments), which are then screened for binding to an antigen of interest.

[0742] In some embodiments, antibody fragments (such as a Fab, a Fab', a F(ab')₂, a scFv, a VH, a VHH, a VNAR, or a diabody) are generated. Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments were derived via proteolytic digestion of intact antibodies. However, these fragments can now be produced directly using recombinant host cells. For example, antibody fragments can be isolated from antibody phage libraries. Alternatively, Fab'-SH fragments can be directly recovered from E. coli cells and chemically coupled to form F(ab')₂ fragments. According to another approach, F(ab')₂ fragments can be isolated directly from recombinant host cell culture. Other techniques for the production of antibody fragments will be apparent to those skilled in the art. [0743] In some embodiments, the antibody or an antibody fragment is conjugated to another molecule, e.g., polyethylene glycol (PEGylation) or serum albumin, to provide an extended half-life in vivo.

Multispecific antibodies

[0744] In some embodiments, multispecific antibodies comprising an anti-alpha-synuclein antibody (or antigen-binding portion thereof) as described herein are provided, e.g., a bispecific antibody. In some embodiments, a multispecific antibody (e.g., a bispecific antibody) has a variable region-binding specificity for alpha-synuclein and has a binding specificity for at least one other antigen. In some embodiments, a multispecific antibody (e.g., a bispecific antibody) binds to two different epitopes of alpha-synuclein.

[0745] In some embodiments, an anti-alpha-synuclein antibody comprises: (a) a first antigen- binding portion comprising a first variable region that specifically binds to an alpha-synuclein protein (e.g., human alpha-synuclein protein), wherein the first antigen-binding portion comprises (i) a first heavy chain comprising a first Fc polypeptide and (ii) a first light chain; and (b) a second antigen-binding portion comprising a second variable region that specifically binds to the alpha-synuclein protein (e.g., human alpha-synuclein protein), wherein the second antigen-binding portion comprises (i) a second heavy chain comprising a second Fc polypeptide and (ii) a second light chain; wherein the first Fc polypeptide and the second Fc polypeptide form an Fc dimer.

[0746] In some embodiments, one variable region (e.g., of a bispecific antibody of the present invention) has a higher binding affinity for monomeric alpha-synuclein protein and/or truncated alpha-synuclein protein than for oligomeric alpha-synuclein protein and/or alpha- synuclein fibrils, and a second variable region has a higher binding affinity for oligomeric alpha-synuclein protein and/or alpha-synuclein fibrils than for monomeric alpha-synuclein protein and/or truncated alpha-synuclein protein.

[0747] In some embodiments, the first Fc polypeptide is a modified Fc polypeptide. In some embodiments, the second Fc polypeptide is a modified Fc polypeptide. In some embodiments, the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide. In some embodiments, the first Fc polypeptide is a modified Fc polypeptide and the second Fc polypeptide is a modified Fc polypeptide. [0748] In some embodiments, the first and second variable regions recognize the same epitope in the alpha-synuclein protein. In some embodiments, the first and second variable regions recognize different epitopes in the alpha-synuclein protein.

[0749] In some embodiments, an anti-alpha-synuclein antibody comprises: (a) a first antigen- binding portion comprising a first variable region that specifically binds to an alpha-synuclein protein (e.g., human alpha-synuclein protein), wherein the first antigen-binding portion comprises (i) a first heavy chain comprising a first Fc polypeptide and (ii) a first light chain; and (b) a second antigen-binding portion comprising a second variable region that specifically binds to the alpha-synuclein protein (e.g., human alpha-synuclein protein), wherein the second antigen-binding portion comprises (i) a second heavy chain comprising a second Fc polypeptide and (ii) a second light chain; wherein the first Fc polypeptide and the second Fc polypeptide form an Fc dimer and wherein the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide

[0750] Methods for making multispecific antibodies include, but are not limited to, recombinant co-expression of two pairs of heavy chain and light chain in a host cell, "knobs- into-holes" engineering, "diabody" technology, intramolecular trimerization, and fusion of an antibody fragment to the N-terminus or C-terminus of another antibody, e.g., tandem variable domains.

Nucleic acids, vectors, and host cells

[0751] In some embodiments, the anti-alpha-synuclein antibodies as described herein are prepared using recombinant methods. Accordingly, in some aspects, the invention provides isolated nucleic acids comprising a nucleic acid sequence encoding any of the anti-alpha- synuclein antibodies as described herein (e.g., any one or more of the CDRs, heavy chain variable regions, and light chain variable regions described herein); vectors comprising such nucleic acids; and host cells into which the nucleic acids are introduced that are used to replicate the antibody-encoding nucleic acids and/or to express the antibodies.

[0752] In some embodiments, a polynucleotide (e.g., an isolated polynucleotide) comprises a nucleotide sequence encoding an antibody or antigen-binding portion thereof as described herein (e.g., as described in the Section above entitled "Anti-Alpha-Synuclein Antibody Sequences"). In some embodiments, the polynucleotide comprises a nucleotide sequence encoding one or more amino acid sequences (e.g., CDR, heavy chain, light chain, and/or framework regions) disclosed in Table 15 below. In some embodiments, the polynucleotide comprises a nucleotide sequence encoding an amino acid sequence having at least 85% sequence identity (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) to a sequence (e.g., a CDR, heavy chain, light chain, or framework region sequence) disclosed in Table 15 below. In some embodiments, a polynucleotide as described herein is operably linked to a heterologous nucleic acid, e.g., a heterologous promoter.

[0753] Suitable vectors containing polynucleotides encoding antibodies of the present disclosure, or fragments thereof, include cloning vectors and expression vectors. While the cloning vector selected may vary according to the host cell intended to be used, useful cloning vectors generally have the ability to self-replicate, may possess a single target for a particular restriction endonuclease, and/or may carry genes for a marker that can be used in selecting clones containing the vector. Examples include plasmids and bacterial viruses, e.g., pUC 18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322, pMB9, ColEl, pCRl, RP4, phage DNAs, and shuttle vectors such as pSA3 and pAT28. These and many other cloning vectors are available from commercial vendors such as BioRad, Strategene, and Invitrogen.

[0754] Expression vectors generally are replicable polynucleotide constructs that contain a nucleic acid of the present disclosure. The expression vector may replicate in the host cells either as episomes or as an integral part of the chromosomal DNA. Suitable expression vectors include but are not limited to plasmids, viral vectors, including adenoviruses, adeno-associated viruses, retroviruses, and any other vector.

[0755] Suitable host cells for cloning or expressing a polynucleotide or vector as described herein include prokaryotic or eukaryotic cells. In some embodiments, the host cell is prokaryotic. In some embodiments, the host cell is eukaryotic, e.g., Chinese Hamster Ovary (CHO) cells or lymphoid cells. In some embodiments, the host cell is a human cell, e.g., a Human Embryonic Kidney (HEK) cell.

[0756] In a further aspect, methods of making an anti-alpha-synuclein antibody as described herein are provided. In some embodiments, the method includes culturing a host cell as described herein (e.g., a host cell expressing a polynucleotide or vector as described herein) under conditions suitable for expression of the antibody. In some embodiments, the antibody is subsequently recovered from the host cell (or host cell culture medium). Hybridoma cell lines

[0757] In another aspect, hybridoma cell lines are provided. Methods of generating and screening hybridoma cell lines, including the selection and immunization of suitable animals, the isolation and fusion of appropriate cells to create the hybridomas, the screening of hybridomas for the secretion of desired antibodies, and characterization of the antibodies will be known to one of ordinary skill in the art. Non-limiting examples are also described in Example 5 herein. In some embodiments, the hybridoma cell line secretes an anti-alpha- synuclein antibody (e.g., monoclonal anti-alpha-synuclein antibody) of the present invention. In some embodiments, the hybridoma cell line is selected from the group consisting of 3D5, 4F5, 46D2, 28B6, 3E12, 1F4, 1G4, and 16D4. In some embodiments, the hybridoma cell is the hybridoma cell line designated 3D5, which secretes the anti-alpha-synuclein antibody 3D5. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 4F5, which secretes the anti-alpha-synuclein antibody 4F5. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 46D2, which secretes the anti-alpha-synuclein antibody 46D2. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 28B6, which secretes the anti-alpha-synuclein antibody 28B6. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 3E12, which secretes the anti-alpha- synuclein antibody 3E12. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 1F4, which secretes the anti-alpha-synuclein antibody 1F4. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 1G4, which secretes the anti-alpha-synuclein antibody 1G4. In some embodiments, the hybridoma cell line is the hybridoma cell line designated 16D4, which secretes the anti-alpha-synuclein antibody 16D4.

[0758] In still another aspect, antibodies (e.g., monoclonal antibodies) secreted by hybridoma cell lines of the present invention are provided. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 3D5, 4F5, 46D2, 28B6, 3E12, 1F4, 1G4, or 16D4. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 3D5. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 4F5. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 46D2. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 28B6. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 3E12. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 1F4. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 1G4. In some embodiments, the antibody is the anti-alpha-synuclein antibody secreted by the hybridoma cell line 16D4.

IV. FC POLYPEPTIDE MODIFICATIONS FOR BLOOD-BRAIN BARRIER (BBB) RECEPTOR BINDING

[0759] In some aspects, provided herein are anti-alpha-synuclein antibodies that are capable of being transported across the blood-brain barrier (BBB). Such a protein comprises a modified Fc polypeptide that binds to a BBB receptor. BBB receptors are expressed on BBB endothelia, as well as other cell and tissue types. In some embodiments, the BBB receptor is a transferrin receptor (TfR).

[0760] Amino acid residues designated in various Fc modifications, including those introduced in a modified Fc polypeptide that binds to a BBB receptor, e.g., TfR, are numbered herein using EU index numbering. Any Fc polypeptide, e.g., an IgGl, IgG2, IgG3, or IgG4 Fc polypeptide, may have modifications, e.g., amino acid substitutions, in one or more positions as described herein.

[0761] In some embodiments, an anti-alpha-synuclein antibody comprises a first and optionally a second Fc polypeptide, each of which can be independently modified. In some embodiments, modifications (e.g., that promote TfR binding) that are made to the first and/or second Fc polypeptides result in an increase in brain uptake of the antibody (or antigen-binding portion thereof) of at least about 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10- fold, 11-fold, 12-fold, 13-fold, 14-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold, 20-fold, or more, compared to the uptake without the modifications having been made.

[0762] A modified (e.g., enhancing heterodimerization and/or BBB receptor-binding) Fc polypeptide can have at least 70% identity, at least 75% identity, at least 80%> identity, at least 85%) identity, at least 90% identity, or at least 95% identity to a native Fc region sequence or a fragment thereof, e.g., a fragment of at least 50 amino acids or at least 100 amino acids, or greater in length. In some embodiments, the native Fc amino acid sequence is the Fc region sequence of SEQ ID NO:455. In some embodiments, the modified Fc polypeptide has at least 70%) identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 1-110 of SEQ ID NO:455, or to amino acids 111-217 of SEQ ID NO:455, or a fragment thereof, e.g., a fragment of at least 50 amino acids or at least 100 amino acids, or greater in length. [0763] In some embodiments, a modified (e.g., enhancing heterodimerization and/or BBB receptor-binding) Fc polypeptide comprises at least 50 amino acids, or at least 60, 65, 70, 75, 80, 85, 90, or 95 or more, or at least 100 amino acids, or more, that correspond to a native Fc region amino acid sequence. In some embodiments, the modified Fc polypeptide comprises at least 25 contiguous amino acids, or at least 30, 35, 40, or 45 contiguous amino acids, or 50 contiguous amino acids, or at least 60, 65, 70, 75, 80 85, 90, or 95 or more contiguous amino acids, or 100 or more contiguous amino acids, that correspond to a native Fc region amino acid sequence, such as SEQ ID NO:455.

[0764] In some embodiments, the domain that is modified for BBB receptor-binding activity is a human Ig CH3 domain, such as an IgGl CH3 domain. The CH3 domain can be of any IgG subtype, i.e., from IgGl, IgG2, IgG3, or IgG4. In the context of IgGl antibodies, a CH3 domain refers to the segment of amino acids from about position 341 to about position 447 as numbered according to the EU numbering scheme.

[0765] In some embodiments, the domain that is modified for BBB receptor-binding activity is a human Ig CH2 domain, such as an IgG CH2 domain. The CH2 domain can be of any IgG subtype, i.e., from IgGl, IgG2, IgG3, or IgG4. In the context of IgGl antibodies, a CH2 domain refers to the segment of amino acids from about position 231 to about position 340 as numbered according to the EU numbering scheme.

[0766] In some embodiments, a modified (e.g., BBB receptor-binding) Fc polypeptide comprises at least one, two, or three substitutions; and in some embodiments, at least four five, six, seven, eight, nine, or ten substitutions at amino acid positions comprising 266, 267, 268, 269, 270, 271, 295, 297, 298, and 299, according to the EU numbering scheme.

[0767] In some embodiments, a modified (e.g., BBB receptor-binding) Fc polypeptide comprises at least one, two, or three substitutions; and in some embodiments, at least four, five, six, seven, eight, or nine substitutions at amino acid positions comprising 274, 276, 283, 285, 286, 287, 288, 289, and 290, according to the EU numbering scheme.

[0768] In some embodiments, a modified (e.g., BBB receptor-binding) Fc polypeptide comprises at least one, two, or three substitutions; and in some embodiments, at least four, five, six, seven, eight, nine, or ten substitutions at amino acid positions comprising 268, 269, 270, 271, 272, 292, 293, 294, 296, and 300, according to the EU numbering scheme. [0769] In some embodiments, a modified (e.g., BBB receptor-binding) Fc polypeptide comprises at least one, two, or three substitutions; and in some embodiments, at least four, five, six, seven, eight, or nine substitutions at amino acid positions comprising 272, 274, 276, 322, 324, 326, 329, 330, and 331, according to the EU numbering scheme.

[0770] In some embodiments, a modified (e.g., BBB receptor-binding) Fc polypeptide comprises at least one, two, or three substitutions; and in some embodiments, at least four, five, six, or seven substitutions at amino acid positions comprising 345, 346, 347, 349, 437, 438, 439, and 440, according to the EU numbering scheme.

[0771] In some embodiments, a modified (e.g., BBB receptor-binding) Fc polypeptide comprises at least one, two, or three substitutions; and in some embodiments, at least four, five, six, seven, eight, or nine substitutions at amino acid positions 384, 386, 387, 388, 389, 390, 413, 416, and 421, according to the EU numbering scheme.

[0772] In some embodiments, an anti-alpha-synuclein antibody comprises two Fc polypeptides, wherein one Fc polypeptide is not modified to bind to a BBB receptor (e.g., TfR) and the other Fc polypeptide is modified to specifically bind to a BBB receptor (e.g., TfR).

FcRn binding sites

[0773] In certain aspects, modified (e.g., BBB receptor-binding) Fc polypeptides, or Fc polypeptides that do not specifically bind to a BBB receptor, can also comprise an FcRn binding site. In some embodiments, the FcRn binding site is within the Fc polypeptide or a fragment thereof.

[0774] In some embodiments, the FcRn binding site comprises a native FcRn binding site. In some embodiments, the FcRn binding site does not comprise amino acid changes relative to the amino acid sequence of a native FcRn binding site. In some embodiments, the native FcRn binding site is an IgG binding site, e.g., a human IgG binding site. In some embodiments, the FcRn binding site comprises a modification that alters FcRn binding.

[0775] In some embodiments, one or more Fc polypeptides (e.g., a first Fc polypeptide, a second Fc polypeptide, or a first and second Fc polypeptide) contain modifications that affect (e.g., increase) FcRn binding. In some embodiments, an FcRn binding site has one or more amino acid residues that are mutated, e.g., substituted, wherein the mutation(s) increase serum half-life or do not substantially reduce serum half-life (i.e., reduce serum half-life by no more than 25% compared to a counterpart modified Fc polypeptide having the wild-type residues at the mutated positions when assayed under the same conditions). In some embodiments, an FcRn binding site has one or more amino acid residues that are substituted at positions 251- 256, 428, and 433-436, according to the EU numbering scheme.

[0776] In some embodiments, one or more residues at or near an FcRn binding site are mutated, relative to a native human IgG sequence, to extend serum half-life of the modified polypeptide. In some embodiments, a mutation, e.g., a substitution, is introduced at one or more of positions 244-257, 279-284, 307-317, 383-390, and 428-435, according to the EU numbering scheme. In some embodiments, one or more mutations are introduced at positions 251, 252, 254, 255, 256, 307, 308, 309, 311, 312, 314, 385, 386, 387, 389, 428, 433, 434, or 436, according to the EU numbering scheme. In some embodiments, mutations are introduced into one, two, or three of positions 252, 254, and 256. In some embodiments, the mutations are M252Y, S254T, and T256E. In some embodiments, a modified Fc polypeptide further comprises the mutations M252Y, S254T, and T256E. In some embodiments, a modified Fc polypeptide comprises a substitution at one, two, or all three of positions T307, E380, and N434, according to the EU numbering scheme. In some embodiments, the mutations are T307Q and N434A. In some embodiments, a modified Fc polypeptide comprises mutations T307A, E380A, and N434A. In some embodiments, a modified Fc polypeptide comprises substitutions at positions T250 and M428, according to the EU numbering scheme. In some embodiments, the modified Fc polypeptide comprises mutations T250Q and/or M428L. In some embodiments, a modified Fc polypeptide comprises mutations at positions M428 and N434, according to the EU numbering scheme. In some embodiments, the modified Fc polypeptide comprises mutations M428L and N434S. In some embodiments, the modified Fc polypeptide comprises an N434S or N434A mutation.

V. TRANSFERRIN RECEPTOR-BINDING FC POLYPEPTIDES

[0777] This section describes generation of modified Fc polypeptides in accordance with the invention that bind to transferrin receptor (TfR) and are capable of being transported across the blood-brain barrier (BBB).

TfR-binding Fc polypeptides comprising mutations in the CH3 domain

[0778] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises substitutions in a CH3 domain. In some embodiments, a modified Fc polypeptide comprises a human Ig CH3 domain, such as an IgG CH3 domain, that is modified for TfR- binding activity. The CH3 domain can be of any IgG subtype, i.e., from IgGl, IgG2, IgG3, or IgG4. In the context of IgG antibodies, a CH3 domain refers to the segment of amino acids from about position 341 to about position 447 as numbered according to the EU numbering scheme.

[0779] In some embodiments, a modified Fc polypeptide that specifically binds to TfR binds to the apical domain of TfR and may bind to TfR without blocking or otherwise inhibiting binding of transferrin to TfR. In some embodiments, binding of transferrin to TfR is not substantially inhibited. In some embodiments, binding of transferrin to TfR is inhibited by less than about 50% (e.g., less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%). In some embodiments, binding of transferrin to TfR is inhibited by less than about 20% (e.g., less than about 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%).

[0780] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises at least two, three, four, five, six, seven, eight, or nine substitutions at positions 384, 386, 387, 388, 389, 390, 413, 416, and 421, according to the EU numbering scheme. Illustrative substitutions that may be introduced at these positions are shown in Tables 12 and 13. In some embodiments, the amino acid at position 388 and/or 421 is an aromatic amino acid, e.g., Tip, Phe, or Tyr. In some embodiments, the amino acid at position 388 is Tip. In some embodiments, the aromatic amino acid at position 421 is Tip or Phe.

[0781] In some embodiments, at least one position as follows is substituted: Leu, Tyr, Met, or Val at position 384; Leu, Thr, His, or Pro at position 386; Val, Pro, or an acidic amino acid at position 387; an aromatic amino acid, e.g. Tip at position 388; Val, Ser, or Ala at position 389; an acidic amino acid, Ala, Ser, Leu, Thr, or Pro at position 413; Thr or an acidic amino acid at position 416; or Trp, Tyr, His, or Phe at position 421. In some embodiments, the modified Fc polypeptide may comprise a conservative substitution, e.g., an amino acid in the same charge grouping, hydrophobicity grouping, side chain ring structure grouping (e.g., aromatic amino acids), or size grouping, and/or polar or non-polar grouping, of a specified amino acid at one or more of the positions in the set. Thus, for example, He may be present at position 384, 386, and/or position 413. In some embodiments, the acidic amino acid at position one, two, or each of positions 387, 413, and 416 is Glu. In other embodiments, the acidic amino acid at one, two or each of positions 387, 413, and 416 is Asp. In some embodiments, two, three, four, five, six, seven, or all eight of positions 384, 386, 387, 388, 389, 413, 416, and 421 have an amino acid substitution as specified in this paragraph. [0782] In some embodiments, an Fc polypeptide that is modified as described in the preceding two paragraphs comprises a native Asn at position 390. In some embodiments, the modified Fc polypeptide comprises Gly, His, Gin, Leu, Lys, Val, Phe, Ser, Ala, or Asp at position 390. In some embodiments, the modified Fc polypeptide further comprises one, two, three, or four substitutions at positions comprising 380, 391, 392, and 415, according to the EU numbering scheme. In some embodiments, Trp, Tyr, Leu, or Gin may be present at position 380. In some embodiments, Ser, Thr, Gin, or Phe may be present at position 391. In some embodiments, Gin, Phe, or His may be present at position 392. In some embodiments, Glu may be present at position 415.

[0783] In certain embodiments, the modified Fc polypeptide comprises two, three, four, five, six, seven, eight, nine, ten, or eleven positions selected from the following: Trp, Leu, or Glu at position 380; Tyr or Phe at position 384; Thr at position 386; Glu at position 387; Trp at position 388; Ser, Ala, Val, or Asn at position 389; Ser or Asn at position 390; Thr or Ser at position 413; Glu or Ser at position 415; Glu at position 416; and/or Phe at position 421. In some embodiments, the modified Fc polypeptide comprises all eleven positions as follows: Trp, Leu, or Glu at position 380; Tyr or Phe at position 384; Thr at position 386; Glu at position 387; Trp at position 388; Ser, Ala, Val, or Asn at position 389; Ser or Asn at position 390; Thr or Ser at position 413; Glu or Ser at position 415; Glu at position 416; and/or Phe at position 421.

[0784] In certain embodiments, the modified Fc polypeptide comprises Leu or Met at position 384; Leu, His, or Pro at position 386; Val at position 387; Trp at position 388; Val or Ala at position 389; Pro at position 413; Thr at position 416; and/or Trp at position 421. In some embodiments, the modified Fc polypeptide further comprises Ser, Thr, Gin, or Phe at position 391. In some embodiments, the modified Fc polypeptide further comprises Trp, Tyr, Leu, or Gin at position 380 and/or Gin, Phe, or His at position 392. In some embodiments, Trp is present at position 380 and/or Gin is present at position 392. In some embodiments, the modified Fc polypeptide does not have a Trp at position 380.

[0785] In other embodiments, the modified Fc polypeptide comprises Tyr at position 384; Thr at position 386; Glu or Val and position 387; Trp at position 388; Ser at position 389; Ser or Thr at position 413; Glu at position 416; and/or Phe at position 421. In some embodiments, the modified Fc polypeptide comprises a native Asn at position 390. In certain embodiments, the modified Fc polypeptide further comprises Trp, Tyr, Leu, or Gin at position 380; and/or Glu at position 415. In some embodiments, the modified Fc polypeptide further comprises Trp at position 380 and/or Glu at position 415.

[0786] In additional embodiments, the modified Fc polypeptide further comprises one, two, or three substitutions at positions comprising 414, 424, and 426, according to the EU numbering scheme. In some embodiments, position 414 is Lys, Arg, Gly, or Pro; position 424 is Ser, Thr, Glu, or Lys; and/or position 426 is Ser, Trp, or Gly.

[0787] In some embodiments, the modified Fc polypeptide comprises one or more of the following substitutions: Trp at position 380; Thr at position 386; Trp at position 388; Val at position 389; Thr or Ser at position 413; Glu at position 415; and/or Phe at position 421, according to the EU numbering scheme.

[0788] In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75%) identity, at least 80%> identity, at least 85%> identity, at least 90% identity, or at least 95% identity to amino acids 111-217 of any one of SEQ ID NOs:336-429 (e.g., SEQ ID NOs:336- 372). In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75%) identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to any one of SEQ ID NOs:336-429 (e.g., SEQ ID NOs:336-372). In some embodiments, the modified Fc polypeptide comprises the amino acids at EU index positions 384-390 and/or 413-421 of any one of SEQ ID NOs:336-429 (e.g., SEQ ID NOs:336-372). In some embodiments, the modified Fc polypeptide comprises the amino acids at EU index positions 380-390 and/or 413-421 of any one of SEQ ID NOs:336-429 (e.g., SEQ ID NOs:336- 372). In some embodiments, the modified Fc polypeptide comprises the amino acids at EU index positions 380-392 and/or 413-426 of any one of SEQ ID NOs:336-429 (e.g., SEQ ID NOs:336-372).

[0789] In some embodiments, the modified Fc polypeptide has at least 75% identity, at least 80%) identity, at least 85% identity, at least 90% identity, or at least 95% identity to any one of SEQ ID NOs:336-429 (e.g., SEQ ID NOs:336-372), and further comprises at least five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen of the positions, numbered according to the EU index, as follows: Trp, Tyr, Leu, Gin, or Glu at position 380; Leu, Tyr, Met, or Val at position 384; Leu, Thr, His, or Pro at position 386; Val, Pro, or an acidic amino acid at position 387; an aromatic amino acid, e.g. Trp, at position 388; Val, Ser, or Ala at position 389; Ser or Asn at position 390; Ser, Thr, Gin, or Phe at position 391; Gin, Phe, or His at position 392; an acidic amino acid, Ala, Ser, Leu, Thr, or Pro at position 413; Lys, Arg, Gly or Pro at position 414; Glu or Ser at position 415; Thr or an acidic amino acid at position 416; Trp, Tyr, His or Phe at position 421 ; Ser, Thr, Glu or Lys at position 424; and Ser, Trp, or Gly at position 426.

[0790] In some embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:336-372. In other embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:336-372, but in which one, two, or three amino acids are substituted.

[0791] In some embodiments, the modified Fc polypeptide comprises additional mutations such as the mutations described in Section VI below, including, but not limited to, a knob mutation (e.g., T366W as numbered with reference to EU numbering), hole mutations (e.g., T366S, L368A, and Y407V as numbered with reference to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered with reference to EU numbering), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered with reference to EU numbering). By way of illustration, SEQ ID NOs:467-538 provide non-limiting examples of modified Fc polypeptides with mutations in the CH3 domain (e.g., clones CH3C.35.20.1, CH3C.35.23.2, CH3C.35.23.3, CH3C.35.23.4, CH3C.35.21.17.2, and CH3C.35.23) comprising one or more of these additional mutations.

[0792] In some embodiments, the modified Fc polypeptide comprises a knob mutation (e.g., T366W as numbered with reference to EU numbering) and has at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:467, 479, 491, 503, 515, and 527. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:467, 479, 491, 503, 515, and 527.

[0793] In some embodiments, the modified Fc polypeptide comprises a knob mutation (e.g., T366W as numbered with reference to EU numbering) and mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered with reference to EU numbering), and has at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:468, 469, 480, 481, 492, 493, 504, 505, 516, 517, 528, and 529. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:468, 469, 480, 481, 492, 493, 504, 505, 516, 517, 528, and 529. [0794] In some embodiments, the modified Fc polypeptide comprises a knob mutation (e.g., T366W as numbered with reference to EU numbering) and mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered with reference to EU numbering), and has at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:470, 482, 494, 506, 518, and 530. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:470, 482, 494, 506, 518, and 530.

[0795] In some embodiments, the modified Fc polypeptide comprises a knob mutation (e.g., T366W as numbered with reference to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered with reference to EU numbering), and mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered with reference to EU numbering), and has at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:471, 472, 483, 484, 495, 496, 507, 508, 519, 520, 531, and 532. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:471, 472, 483, 484, 495, 496, 507, 508, 519, 520, 531, and 532.

[0796] In some embodiments, the modified Fc polypeptide comprises hole mutations (e.g., T366S, L368A, and Y407V as numbered with reference to EU numbering) and has at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:473, 485, 497, 509, 521, and 533. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:473, 485, 497, 509, 521, and 533.

[0797] In some embodiments, the modified Fc polypeptide comprises hole mutations (e.g., T366S, L368A, and Y407V as numbered with reference to EU numbering) and mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered with reference to EU numbering), and has at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:474, 475, 486, 487, 498, 499, 510, 51 1, 522, 523, 534, and 535. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:474, 475, 486, 487, 498, 499, 510, 51 1, 522, 523, 534, and 535.

[0798] In some embodiments, the modified Fc polypeptide comprises hole mutations (e.g., T366S, L368A, and Y407V as numbered with reference to EU numbering) and mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered with reference to EU numbering), and has at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:476, 488, 500, 512, 524, and 536. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:476, 488, 500, 512, 524, and 536.

[0799] In some embodiments, the modified Fc polypeptide comprises hole mutations (e.g., T366S, L368A, and Y407V as numbered with reference to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered with reference to EU numbering), and mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered with reference to EU numbering), and has at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:477, 478, 489, 490, 501, 502, 513, 514, 525, 526, 537, and 538. In some embodiments, the modified Fc polypeptide comprises the sequence of any one of SEQ ID NOs:477, 478, 489, 490, 501, 502, 513, 514, 525, 526, 537, and 538.

[0800] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises at least two, three, four, five, six, seven, or eight substitutions at positions 345, 346, 347, 349, 437, 438, 439, and 440, according to the EU numbering scheme. Illustrative modified Fc polypeptides are provided in SEQ ID NOs:430-434. In some embodiments, the modified Fc polypeptide comprises Gly at position 437; Phe at position 438; and/or Asp at position 440. In some embodiments, Glu is present at position 440. In certain embodiments, the modified Fc polypeptide comprises at least one substitution at a position as follows: Phe or He at position 345; Asp, Glu, Gly, Ala, or Lys at position 346; Tyr, Met, Leu, He, or Asp at position 347; Thr or Ala at position 349; Gly at position 437; Phe at position 438; His Tyr, Ser, or Phe at position 439; or Asp at position 440. In some embodiments, two, three, four, five, six, seven, or all eight of positions 345, 346, 347, 349, 437, 438, 439, and 440 and have a substitution as specified in this paragraph. In some embodiments, the modified Fc polypeptide may comprise a conservative substitution, e.g., an amino acid in the same charge grouping, hydrophobicity grouping, side chain ring structure grouping (e.g., aromatic amino acids), or size grouping, and/or polar or non-polar grouping, of a specified amino acid at one or more of the positions in the set.

[0801] In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75%) identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 111-217 of any one of SEQ ID NOs:430-434. In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to SEQ ID NOs:430-434. In some embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:430-434. In other embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:430-434, but in which one, two, or three amino acids are substituted.

TfR-binding Fc polypeptides comprising mutations in the CH2 domain

[0802] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises substitutions in a CH2 domain. In some embodiments, a modified Fc polypeptide comprises a human Ig CH2 domain, such as an IgG CH2 domain, that is modified for TfR- binding activity. The CH2 domain can be of any IgG subtype, i.e., from IgGl, IgG2, IgG3, or IgG4. In the context of IgG antibodies, a CH2 domain refers to the segment of amino acids from about position 231 to about position 340 as numbered according to the EU numbering scheme.

[0803] In some embodiments, a modified Fc polypeptide that specifically binds to TfR binds to the apical domain of TfR and may bind to TfR without blocking or otherwise inhibiting binding of transferrin to TfR. In some embodiments, binding of transferrin to TfR is not substantially inhibited. In some embodiments, binding of transferrin to TfR is inhibited by less than about 50% (e.g., less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%). In some embodiments, binding of transferrin to TfR is inhibited by less than about 20% (e.g., less than about 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%).

[0804] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises at least two, three, four, five, six, seven, eight, or nine substitutions at positions 274, 276, 283, 285, 286, 287, 288, and 290, according to the EU numbering scheme. Illustrative modified Fc polypeptides are provided in SEQ ID NOs:435-439. In some embodiments, the modified Fc polypeptide comprises Glu at position 287 and/or Trp at position 288. In some embodiments, the modified Fc polypeptide comprises at least one substitution at a position as follows: Glu, Gly, Gin, Ser, Ala, Asn, Tyr, or Trp at position 274; He, Val, Asp, Glu, Thr, Ala, or Tyr at position 276; Asp, Pro, Met, Leu, Ala, Asn, or Phe at position 283; Arg, Ser, Ala, or Gly at position 285; Tyr, Trp, Arg, or Val at position 286; Glu at position 287; Trp or Tyr at position 288; Gin, Tyr, His, He, Phe, Val, or Asp at position 289; or Leu, Trp, Arg, Asn, Tyr, or Val at position 290. In some embodiments, two, three, four, five, six, seven, eight, or all nine of positions 274, 276, 283, 285, 286, 287, 288, and 290 have a substitution as specified in this paragraph. In some embodiments, the modified Fc polypeptide may comprise a conservative substitution, e.g., an amino acid in the same charge grouping, hydrophobicity grouping, side chain ring structure grouping (e.g., aromatic amino acids), or size grouping, and/or polar or non-polar grouping, of a specified amino acid at one or more of the positions in the set.

[0805] In some embodiments, the modified Fc polypeptide comprises Glu, Gly, Gin, Ser, Ala, Asn, or Tyr at position 274; He, Val, Asp, Glu, Thr, Ala, or Tyr at position 276 Asp, Pro, Met, Leu, Ala, or Asn at position 283; Arg, Ser, or Ala at position 285; Tyr, Trp, Arg, or Val at position 286; Glu at position 287; Trp at position 288; Gin, Tyr, His, He, Phe, or Val at position 289; and/or Leu, Trp, Arg, Asn, or Tyr at position 290. In some embodiments, the modified Fc polypeptide comprises Arg at position 285; Tyr or Trp at position 286; Glu at position 287; Trp at position 288; and/or Arg or Trp at position 290.

[0806] In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 1-110 of any one of SEQ ID NOs:435-439. In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to SEQ ID NOs:435-439. In some embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:435-439. In other embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:435-439, but in which one, two, or three amino acids are substituted.

[0807] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises at least two, three, four, five, six, seven, eight, nine, or ten substitutions at positions 266, 267, 268, 269, 270, 271, 295, 297, 298, and 299, according to the EU numbering scheme. Illustrative modified Fc polypeptides are provided in SEQ ID NOs:440-444. In some embodiments, the modified Fc polypeptide comprises Pro at position 270, Glu at position 295, and/or Tyr at position 297. In some embodiments, the modified Fc polypeptide comprises at least one substitution at a position as follows: Pro, Phe, Ala, Met, or Asp at position 266; Gin, Pro, Arg, Lys, Ala, He, Leu, Glu, Asp, or Tyr at position 267; Thr, Ser, Gly, Met, Val, Phe, Trp, or Leu at position 268; Pro, Val, Ala, Thr, or Asp at position 269; Pro, Val, or Phe at position 270; Trp, Gin, Thr, or Glu at position 271; Glu, Val, Thr, Leu, or Tip at position 295; Tyr, His, Val, or Asp at position 297; Thr, His, Gin, Arg, Asn, or Val at position 298; or Tyr, Asn, Asp, Ser, or Pro at position 299. In some embodiments, two, three, four, five, six, seven, eight, nine, or all ten of positions 266, 267, 268, 269, 270, 271, 295, 297, 298, and 299 have a substitution as specified in this paragraph. In some embodiments, a modified Fc polypeptide may comprise a conservative substitution, e.g., an amino acid in the same charge grouping, hydrophobicity grouping, side chain ring structure grouping (e.g., aromatic amino acids), or size grouping, and/or polar or non-polar grouping, of a specified amino acid at one or more of the positions in the set.

[0808] In some embodiments, the modified Fc polypeptide comprises Pro, Phe, or Ala at position 266; Gin, Pro, Arg, Lys, Ala, or He at position 267; Thr, Ser, Gly, Met, Val, Phe, or Trp at position 268; Pro, Val, or Ala at position 269; Pro at position 270; Trp or Gin at position 271; Glu at position 295; Tyr at position 297; Thr, His, or Gin at position 298; and/or Tyr, Asn, Asp, or Ser at position 299.

[0809] In some embodiments, the modified Fc polypeptide comprises Met at position 266; Leu or Glu at position 267; Trp at position 268; Pro at position 269; Val at position 270; Thr at position 271; Val or Thr at position 295; His at position 197; His, Arg, or Asn at position 198; and/or Pro at position 299.

[0810] In some embodiments, the modified Fc polypeptide comprises Asp at position 266; Asp at position 267; Leu at position 268; Thr at position 269; Phe at position 270; Gin at position 271; Val or Leu at position 295; Val at position 297; Thr at position 298; and/or Pro at position 299.

[0811] In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 1-110 of any one of SEQ ID NOs:440-444. In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to SEQ ID NOs:440-444. In some embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:440-444. In other embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:440-444, but in which one, two, or three amino acids are substituted. [0812] In some embodiments, a modified Fc polypeptide that specifically binds to TfR comprises at least two, three, four, five, six, seven, eight, nine, or ten substitutions at positions 268, 269, 270, 271, 272, 292, 293, 294, and 300, according to the EU numbering scheme. Illustrative modified Fc polypeptides are provided in SEQ ID NOs:445-449. In some embodiments, the modified Fc polypeptide comprises at least one substitution at a position as follows: Val or Asp at position 268; Pro, Met, or Asp at position 269; Pro or Trp at position 270; Arg, Trp, Glu, or Thr at position 271; Met, Tyr, or Trp at position 272; Leu or Trp at position 292; Thr, Val, He, or Lys at position 293; Ser, Lys, Ala, or Leu at position 294; His, Leu, or Pro at position 296; or Val or Trp at position 300. In some embodiments, two, three, four, five, six, seven, eight, nine, or all ten of positions 268, 269, 270, 271, 272, 292, 293, 294, and 300 have a substitution as specified in this paragraph. In some embodiments, the modified Fc polypeptide may comprise a conservative substitution, e.g., an amino acid in the same charge grouping, hydrophobicity grouping, side chain ring structure grouping (e.g., aromatic amino acids), or size grouping, and/or polar or non-polar grouping, of a specified amino acid at one or more of the positions in the set.

[0813] In some embodiments, the modified Fc polypeptide comprises Val at position 268; Pro at position 269; Pro at position 270; Arg or Trp at position 271; Met at position 272; Leu at position 292; Thr at position 293; Ser at position 294; His at position 296; and/or Val at position 300.

[0814] In some embodiments, the modified Fc polypeptide comprises Asp at position 268; Met or Asp at position 269; Trp at position 270; Glu or Thr at position 271; Tyr or Trp at position 272; Trp at position 292; Val, He, or Lys at position 293; Lys, Ala, or Leu at position 294; Leu or Pro at position 296; and/or Trp at position 300.

[0815] In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 1-110 of any one of SEQ ID NOs:445-449. In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to SEQ ID NOs:445-449. In some embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:445-449. In other embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:445-449, but in which one, two, or three amino acids are substituted. [0816] In some embodiments, a modified Fc polypeptide that specifically binds to TfR has at least two, three, four, five, six, seven, eight, nine, or ten substitutions at positions 272, 274, 276, 322, 324, 326, 329, 330, and 331, according to the EU numbering scheme. An illustrative modified polypeptide comprises Trp at position 330. In some embodiments, the modified Fc polypeptide comprises at least one substitution at a position as follows: Trp, Val, He, or Ala at position 272; Trp or Gly at position 274; Tyr, Arg, or Glu at position 276; Ser, Arg, or Gin at position 322; Val, Ser, or Phe at position 324; He, Ser, or Trp at position 326; Trp, Thr, Ser, Arg, or Asp at position 329; Trp at position 330; or Ser, Lys, Arg, or Val at position 331. In some embodiments, two, three, four, five, six, seven, eight, or all nine of positions 272, 274, 276, 322, 324, 326, 329, 330, and 331 have a substitution as specified in this paragraph. In some embodiments, the modified Fc polypeptide may comprise a conservative substitution, e.g., an amino acid in the same charge grouping, hydrophobicity grouping, side chain ring structure grouping (e.g., aromatic amino acids), or size grouping, and/or polar or non-polar grouping, of a specified amino acid at one or more of the positions in the set.

[0817] In some embodiments, the modified Fc polypeptide comprises two, three, four, five, six, seven, eight, or nine positions selected from the following: position 272 is Trp, Val, He, or Ala; position 274 is Trp or Gly; position 276 is Tyr, Arg, or Glu; position 322 is Ser, Arg, or Gin; position 324 is Val, Ser, or Phe; position 326 is He, Ser, or Trp; position 329 is Trp, Thr, Ser, Arg, or Asp; position 330 is Trp; and position 331 is Ser, Lys, Arg, or Val. In some embodiments, the modified Fc polypeptide comprises Val or He at position 272; Gly at position 274; Arg at position 276; Arg at position 322; Ser at position 324; Ser at position 326; Thr, Ser, or Arg at position 329; Trp at position 330; and/or Lys or Arg at position 331.

[0818] In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 1-110 of any one of SEQ ID NOs:450-454. In some embodiments, the modified Fc polypeptide has at least 70% identity, at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, or at least 95% identity to SEQ ID NOs:450-454. In some embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:450-454. In other embodiments, the modified Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:450-454, but in which one, two, or three amino acids are substituted. VI. ADDITIONAL FC POLYPEPTIDE MUTATIONS

[0819] In some aspects, an anti-alpha-synuclein antibody of the invention comprises first and optionally second Fc polypeptides that may each comprise independently selected modifications or may be a wild-type Fc polypeptide, e.g., a human IgGl Fc polypeptide. In some embodiments, one or both Fc polypeptides contains one or more modifications that confer binding to a blood-brain barrier (BBB) receptor, e.g., transferrin receptor (TfR). Non-limiting examples of other mutations that can be introduced into one or both Fc polypeptides include, e.g., mutations to increase serum stability, to modulate effector function, to influence glycosylation, to reduce immunogenicity in humans, and/or to provide for knob and hole heterodimerization of the Fc polypeptides.

[0820] In some embodiments, one or more Fc polypeptides (e.g., a first and optionally a second Fc polypeptide) has an amino acid sequence identity of at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%), 95%), 96%), 97%), 98%, or 99% to a corresponding wild-type Fc polypeptide (e.g., a human IgGl, IgG2, IgG3, or IgG4 Fc polypeptide).

[0821] In some embodiments, the Fc polypeptides include knob and hole mutations to promote heterodimer formation and hinder homodimer formation. Generally, the modifications introduce a protuberance ("knob") at the interface of a first polypeptide and a corresponding cavity ("hole") in the interface of a second polypeptide, such that the protuberance can be positioned in the cavity so as to promote heterodimer formation and thus hinder homodimer formation. Protuberances are constructed by replacing small amino acid side chains from the interface of the first polypeptide with larger side chains (e.g., tyrosine or tryptophan). Compensatory cavities of identical or similar size to the protuberances are created in the interface of the second polypeptide by replacing large amino acid side chains with smaller ones (e.g., alanine or threonine). In some embodiments, such additional mutations are at a position in the Fc polypeptide that does not have a negative effect on binding of the polypeptide to a BBB receptor, e.g., TfR.

[0822] In one illustrative embodiment of a knob and hole approach for dimerization, position 366 (numbered according to the EU numbering scheme) of one of the Fc polypeptides comprises a tryptophan in place of a native threonine. The other Fc polypeptide in the dimer has a valine at position 407 (numbered according to the EU numbering scheme) in place of the native tyrosine. The other Fc polypeptide may further comprise a substitution in which the native threonine at position 366 (numbered according to the EU numbering scheme) is substituted with a serine and a native leucine at position 368 (numbered according to the EU numbering scheme) is substituted with an alanine. Thus, one of the Fc polypeptides of has the T366W knob mutation and the other Fc polypeptide has the Y407V mutation, which is typically accompanied by the T366S and L368A hole mutations.

[0823] In some embodiments, modifications to enhance serum half-life may be introduced. For example, in some embodiments, one or both Fc polypeptides may comprise a tyrosine at position 252, a threonine at position 254, and a glutamic acid at position 256, as numbered according to the EU numbering scheme. Thus, one or both Fc polypeptides may have M252Y, S254T, and T256E substitutions. Alternatively, one or both Fc polypeptides may have M428L and N434S substitutions. Alternatively, one or both Fc polypeptides may have an N434S or N434A substitution.

[0824] In some embodiments, one or both Fc polypeptides may comprise modifications that reduce effector function, i.e., having a reduced ability to induce certain biological functions upon binding to an Fc receptor expressed on an effector cell that mediates the effector function. Examples of antibody effector functions include, but are not limited to, Clq binding and complement dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell- mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), down-regulation of cell surface receptors {e.g., B cell receptor), and B-cell activation. Effector functions may vary with the antibody class. For example, native human IgGl and IgG3 antibodies can elicit ADCC and CDC activities upon binding to an appropriate Fc receptor present on an immune system cell; and native human IgGl, IgG2, IgG3, and IgG4 Fc can elicit ADCP functions upon binding to the appropriate Fc receptor present on an immune cell.

[0825] In some embodiments, one or both Fc polypeptides may also be engineered to contain other modifications for heterodimerization, e.g., electrostatic engineering of contact residues within a CH3-CH3 interface that are naturally charged or hydrophobic patch modifications.

[0826] In some embodiments, one or both Fc polypeptides may include additional modifications that modulate effector function.

[0827] In some embodiments, one or both Fc polypeptides may comprise modifications that reduce or eliminate effector function. Illustrative Fc polypeptide mutations that reduce effector function include, but are not limited to, substitutions in a CH2 domain, e.g., at positions 234 and 235, according to the EU numbering scheme. For example, in some embodiments, one or both Fc polypeptides can comprise alanine residues at positions 234 and 235. Thus, one or both Fc polypeptides may have L234A and L235A (LALA) substitutions.

[0828] Additional Fc polypeptide mutations that modulate an effector function include, but are not limited to, one or more substitutions at positions 238, 265, 269, 270, 297, 327 and 329, according to the EU numbering scheme. Illustrative substitutions include the following: position 329 may have a mutation in which proline is substituted with a glycine or arginine or an amino acid residue large enough to destroy the Fc/Fcy receptor interface that is formed between proline 329 of the Fc and tryptophan residues Tip 87 and Tip 110 of FcyRIII. Additional illustrative substitutions include S228P, E233P, L235E, N297A, N297D, and P331 S, according to the EU numbering scheme. Multiple substitutions may also be present, e.g., L234A and L235A of a human IgGl Fc region; L234A, L235A, and P329G of a human IgGl Fc region; S228P and L235E of a human IgG4 Fc region; L234A and G237A of a human IgGl Fc region; L234A, L235A, and G237A of a human IgGl Fc region; V234A and G237A of a human IgG2 Fc region; L235A, G237A, and E318A of a human IgG4 Fc region; and S228P and L236E of a human IgG4 Fc region, according to the EU numbering scheme. In some embodiments, one or both Fc polypeptides may have one or more amino acid substitutions that modulate ADCC, e.g., substitutions at positions 298, 333, and/or 334, according to the EU numbering scheme.

Illustrative Fc polypeptides comprising additional mutations

[0829] By way of non-limiting example, one or both Fc polypeptides present in an anti- alpha-synuclein antibody of the invention may comprise additional mutations including a knob mutation (e.g., T366W as numbered according to the EU numbering scheme), hole mutations (e.g., T366S, L368A, and Y407V as numbered according to the EU numbering scheme), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered according to the EU numbering scheme), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered according to the EU numbering scheme).

[0830] In some embodiments, an Fc polypeptide may have a knob mutation (e.g., T366W as numbered according to the EU numbering scheme) and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430- 455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have a knob mutation. [0831] In some embodiments, an Fc polypeptide may have a knob mutation (e.g., T366W as numbered according to the EU numbering scheme), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered according to the EU numbering scheme), and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430-455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have a knob mutation and mutations that modulate effector function.

[0832] In some embodiments, an Fc polypeptide may have a knob mutation (e.g., T366W as numbered according to the EU numbering scheme), mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered according to the EU numbering scheme), and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430-455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have a knob mutation and mutations that increase serum stability.

[0833] In some embodiments, an Fc polypeptide may have a knob mutation (e.g., T366W as numbered according to the EU numbering scheme), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered according to the EU numbering scheme), mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered according to the EU numbering scheme), and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430-455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have a knob mutation, mutations that modulate effector function, and mutations that increase serum stability.

[0834] In some embodiments, an Fc polypeptide may have hole mutations (e.g., T366S, L368A, and Y407V as numbered according to the EU numbering scheme) and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430-455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have hole mutations.

[0835] In some embodiments, an Fc polypeptide may have hole mutations (e.g., T366S, L368A, and Y407V as numbered according to the EU numbering scheme), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered according to the EU numbering scheme), and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430- 455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have hole mutations and mutations that modulate effector function.

[0836] In some embodiments, an Fc polypeptide may have hole mutations (e.g., T366S, L368A, and Y407V as numbered according to the EU numbering scheme), mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered according to the EU numbering scheme), and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430-455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have hole mutations and mutations that increase serum stability.

[0837] In some embodiments, an Fc polypeptide may have hole mutations (e.g., T366S, L368A, and Y407V as numbered according to the EU numbering scheme), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) as numbered according to the EU numbering scheme), mutations that increase serum stability (e.g., M252Y, S254T, and T256E as numbered according to the EU numbering scheme), and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:336-421 and 430-455. In some embodiments, an Fc polypeptide having the sequence of any one of SEQ ID NOs:336-421 and 430-455 may be modified to have hole mutations, mutations that modulate effector function, and mutations that increase serum stability.

VII. ANTI- ALPHA- S YNUCLEIN ANTIBODIES HAVING MODIFIED FC POLYPEPTIDES

[0838] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:221, 229, and 238 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:617, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 1 16, and 142, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for the sequence of SEQ ID NO: 544, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 1 16, and 142, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for the sequence of SEQ ID NO:561. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:617, a first heavy chain having the sequence of SEQ ID NO: 544, and a second heavy chain having the sequence of SEQ ID NO:561.

[0839] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:221, 229, and 238 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:617, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:332 and 543-547 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:334 and 560-564 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti- alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:617, a first heavy chain having the sequence of any one of SEQ ID NOs:332 and 543-547, and a second heavy chain having the sequence of any one of SEQ ID NOs:334 and 560-564.

[0840] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:221, 229, and 238 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:617, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:548-553 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:554-559 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti-alpha- synuclein antibody comprises a light chain having the sequence of SEQ ID NO:617, a first heavy chain having the sequence of any one of SEQ ID NOs:548-553, and a second heavy chain having the sequence of any one of SEQ ID NOs:554-559.

[0841] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:221, 229, and 238 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:617, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:332 and 543-547 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:548-553 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti-alpha- synuclein antibody comprises a light chain having the sequence of SEQ ID NO:617, a first heavy chain having the sequence of any one of SEQ ID NOs:332 and 543-547, and a second heavy chain having the sequence of any one of SEQ ID NOs:548-553.

[0842] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:221, 229, and 238 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:617, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:554-559 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 116, and 142, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:334 and 560-564 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:617, a first heavy chain having the sequence of any one of SEQ ID NOs:554-559, and a second heavy chain having the sequence of any one of SEQ ID NOs:334 and 560-564.

[0843] In further embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:221, 229, and 238 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:617, and a heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:79, 1 16, and 142 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:587-592. In some embodiments, the heavy chain comprises a modified Fc polypeptide having a knob mutation (e.g., T366W according to EU numbering), hole mutations (e.g., T366S, L368A, and Y407V according to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) according to EU numbering), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E according to EU numbering).

[0844] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:587-592 may be modified to contain the knob mutation T366W, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0845] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:587-592 may be modified to contain hole mutations T366S, L368A, and Y407V, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0846] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:304, 223, and 308 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:618, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for the sequence of SEQ ID NO:566, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for the sequence of SEQ ID NO: 583. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:618, a first heavy chain having the sequence of SEQ ID NO:566, and a second heavy chain having the sequence of SEQ ID NO: 583.

[0847] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:304, 223, and 308 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:618, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:333 and 565-569 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235 A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:335 and 582-586 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:618, a first heavy chain having the sequence of any one of SEQ ID NOs:333 and 565-569, and a second heavy chain having the sequence of any one of SEQ ID NOs:335 and 582-586.

[0848] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:304, 223, and 308 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:618, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:570-575 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:576-581 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti-alpha- synuclein antibody comprises a light chain having the sequence of SEQ ID NO:618, a first heavy chain having the sequence of any one of SEQ ID NOs: 570-575, and a second heavy chain having the sequence of any one of SEQ ID NOs:576-581.

[0849] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:304, 223, and 308 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:618, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:333 and 565-569 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235 A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:570-575 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti- alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:618, a first heavy chain having the sequence of any one of SEQ ID NOs:333 and 565-569, and a second heavy chain having the sequence of any one of SEQ ID NOs: 570-575.

[0850] In some embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:304, 223, and 308 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:618, a first heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, knob (T366W according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:576-581 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence, and a second heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291, hole (T366S, L368A, and Y407V according to EU numbering), LALA (L234A and L235A according to EU numbering), P329G, and/or YTE (M252Y, S254T, and T256E according to EU numbering) mutations as specified for each sequence of any one of SEQ ID NOs:335 and 582-586 and at least 85% identity, at least 90% identity, or at least 95% identity to the respective sequence. In some embodiments, an anti- alpha-synuclein antibody comprises a light chain having the sequence of SEQ ID NO:618, a first heavy chain having the sequence of any one of SEQ ID NOs:576-581, and a second heavy chain having the sequence of any one of SEQ ID NOs:335 and 582-586.

[0851] In further embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:304, 223, and 308 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:618, and a heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:277, 283, and 291 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:593-598. In some embodiments, the heavy chain comprises a modified Fc polypeptide having a knob mutation (e.g., T366W according to EU numbering), hole mutations (e.g., T366S, L368A, and Y407V according to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) according to EU numbering), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E according to EU numbering).

[0852] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:593-598 may be modified to contain the knob mutation T366W, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0853] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:593-598 may be modified to contain hole mutations T366S, L368A, and Y407V, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0854] In further embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:206, 224, and 232 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO:619, and a heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:278, 284, and 290 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs: 599-604. In some embodiments, the heavy chain comprises a modified Fc polypeptide having a knob mutation (e.g., T366W according to EU numbering), hole mutations (e.g., T366S, L368A, and Y407V according to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) according to EU numbering), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E according to EU numbering).

[0855] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:599-604 may be modified to contain the knob mutation T366W, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0856] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:599-604 may be modified to contain hole mutations T366S, L368A, and Y407V, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0857] In further embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:218, 228, and 237 and at least 85%) identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO: 620, and a heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:65, 88, and 126 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:605-610. In some embodiments, the heavy chain comprises a modified Fc polypeptide having a knob mutation (e.g., T366W according to EU numbering), hole mutations (e.g., T366S, L368A, and Y407V according to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) according to EU numbering), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E according to EU numbering).

[0858] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:605-610 may be modified to contain the knob mutation T366W, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E. [0859] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:605-610 may be modified to contain hole mutations T366S, L368A, and Y407V, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0860] In further embodiments, an anti-alpha-synuclein antibody comprises a light chain having the light chain CDRl-3 sequences set forth in SEQ ID NOs:219, 230, and 237 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of SEQ ID NO: 621, and a heavy chain having the heavy chain CDRl-3 sequences set forth in SEQ ID NOs:65, 1 18, and 144 and at least 85% identity, at least 90% identity, or at least 95% identity to the sequence of any one of SEQ ID NOs:61 1-616. In some embodiments, the heavy chain comprises a modified Fc polypeptide having a knob mutation (e.g., T366W according to EU numbering), hole mutations (e.g., T366S, L368A, and Y407V according to EU numbering), mutations that modulate effector function (e.g., L234A, L235A, and/or P329G (e.g., L234A and L235A) according to EU numbering), and/or mutations that increase serum stability (e.g., M252Y, S254T, and T256E according to EU numbering).

[0861] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:61 1-616 may be modified to contain the knob mutation T366W, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

[0862] In particular embodiments, a heavy chain having the sequence of any one of SEQ ID NOs:611-616 may be modified to contain hole mutations T366S, L368A, and Y407V, mutations that modulate effector function L234A, L235A, and/or P329G (e.g., L234A and L235A), and/or mutations that increase serum stability M252Y, S254T, and T256E.

VIII. THERAPEUTIC METHODS USING ANTI-ALPHA-SYNUCLEIN ANTIBODIES

[0863] In another aspect, methods for the use of anti-alpha-synuclein antibodies as described herein are provided. In some embodiments, an anti-alpha-synuclein antibody as described in Section III above is used in the practice of the methods described herein.

[0864] In some embodiments, methods of neutralizing human alpha-synuclein protein are provided. In some embodiments, the method comprises neutralizing human alpha-synuclein protein in a subject, e.g., in a brain of a subject. In some embodiments, the method comprises administering to the subject an anti-alpha-synuclein antibody or antigen-binding portion thereof as described herein, a pharmaceutical composition comprising an anti-alpha-synuclein antibody as described herein, a multispecific (e.g., bispecific) antibody comprising an anti- alpha-synuclein antibody as described herein, an antibody that competes for binding with an isolated antibody as described herein, or an antibody secreted by a hybridoma cell line as described herein. In some embodiments, the subject is an individual having a neurodegenerative disease (e.g., a synucleinopathy).

[0865] In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting alpha-synuclein oligomerization and/or aggregation in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha- synuclein protein comprises preventing, reducing, or inhibiting alpha-synuclein oligomerization in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting alpha- synuclein aggregation in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting alpha-synuclein oligomerization and aggregation in a subject, e.g., in a brain of a subject.

[0866] In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting the spreading of alpha-synuclein protein oligomers and/or fibrils in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting the spreading of alpha- synuclein protein oligomers in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting the spreading of alpha-synuclein protein fibrils in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting the spreading of alpha-synuclein protein oligomers and fibrils in a subject, e.g., in a brain of a subject.

[0867] In some embodiments, the anti-alpha-synuclein antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody neutralizes monomeric human alpha- synuclein, oligomeric human alpha-synuclein, soluble human alpha-synuclein, and/or human alpha-synuclein fibrils. In some embodiments, the anti-alpha-synuclein antibody or multispecific antibody is a pan-alpha-synuclein antibody or multispecific antibody.

[0868] In some embodiments, methods of treating a neurodegenerative disease (e.g., a synucleinopathy or tauopathy) are provided. In some embodiments, the method comprises administering to a subject having a neurodegenerative disease an anti-alpha-synuclein antibody or antigen-binding portion thereof as described herein, a pharmaceutical composition comprising an anti-alpha-synuclein antibody as described herein, a multispecific (e.g., bispecific) antibody comprising an anti-alpha-synuclein antibody as described herein, an antibody that competes for binding with an isolated antibody as described herein, or an antibody secreted by a hybridoma cell line as described herein.

[0869] In some embodiments, the neurodegenerative disease is characterized by the presence of Lewy bodies and/or Lewy neurites. In some embodiments, the presence of alpha-synuclein accumulation or aggregation (e.g., alpha-synuclein oligomers and/or fibrils), Lewy bodies and/or Lewy neurites are identified in a subject (e.g., before a subject received treatment according to the methods described herein).

[0870] In some embodiments, the neurodegenerative disease is selected from the group consisting of Parkinson' s disease, dementia with Lewy bodies, multiple system atrophy, Huntington' s disease, Alzheimer' s disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down' s syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden-Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle-predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick's disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia.

[0871] In some embodiments, a synucleinopathy is treated. In some embodiments, the synucleinopathy is selected from the group consisting of Parkinson' s disease, dementia with Lewy bodies, and multiple system atrophy.

[0872] In some embodiments, treating the subject delays the onset of symptoms of the disease, causes the reversal of disease symptoms, delays or slows the progression of disease symptoms, lessons the severity of disease symptoms, increases disease survival, or a combination thereof. As non-limiting examples, symptoms of a neurodegenerative disease include memory deficits, cognitive deficits, motor deficits, sensory deficits, and speech deficits.

[0873] In some embodiments, the subject to be treated is a human, e.g., a human adult or a human child.

[0874] In some embodiments, the anti-alpha-synuclein antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody is administered to a subject (e.g., a human subject) prior to the broad spreading of alpha-synuclein protein (e.g., human alpha- synuclein protein) in the brain of the subject. In some embodiments, the anti-alpha-synuclein antibody, pharmaceutical composition, or multispecific antibody is administered to a subject prior to the broad spreading of alpha-synuclein protein oligomers and/or fibrils in the brain of the subject. In some embodiments, the anti-alpha-synuclein antibody, pharmaceutical composition or multispecific antibody is administered to the subject (e.g., to treat a disease such as a neurodegenerative disease in the subject) prior to the broad spreading of alpha- synuclein protein (e.g., alpha-synuclein protein oligomers and/or fibrils) in the brain of the subject, thereby increasing the ability of the isolated antibody, pharmaceutical composition, or bispecific antibody to treat the disease. In some embodiments, treating a subject with the anti- alpha-synuclein antibody, pharmaceutical composition, or multispecific antibody prior to broad spreading of alpha-synuclein protein prevents or delays the onset of symptoms of the disease, causes the reversal of disease symptoms, delays or slows the progression of disease symptoms, lessons the severity of disease symptoms, increases disease survival, or a combination thereof. As non-limiting examples, symptoms of a neurodegenerative disease include memory deficits, cognitive deficits, motor deficits, sensory deficits, and speech deficits.

[0875] In some embodiments, the subject is treated (i.e., the anti-alpha-synuclein antibody, pharmaceutical composition, or multispecific antibody is administered to the subject) during an early stage of the disease (e.g., neurodegenerative disease). One of ordinary skill in the art will appreciate that various diseases progress at different rates and exhibit different combinations of signs and symptoms at each stage. As such, the stages of the various diseases are classified according to different systems. In some diseases, such as Parkinson's disease, the early stages can be presymptomatic. Accordingly, in some embodiments, the term "early stage" as used herein refers to when the subject has not yet exhibited any signs or symptoms of the disease to be treated. In other diseases, such as amyotrophic lateral sclerosis (ALS), early stages of the disease are characterized by symptoms that are limited to a single body region or mild symptoms that affect more than one region. Accordingly, in some embodiments, the term "early stage" as used herein refers to when the subject has one or more symptoms of the disease to be treated that affect a localized region of the body, or when the subject has one or more mild symptoms of the disease to be treated that are not localized to one region of the body. Depending on the particular disease to be treated, the skilled practitioner will be able to determine whether the subject is in an early stage of the disease.

[0876] In some embodiments, the subject has not yet been treated for the disease. In some embodiments, the subject has not been treated with a dopaminergic replacement therapy. In some embodiments, the subject has not been treated with a dopamine agonist therapy.

[0877] In some embodiments, the amount of the isolated antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody that is necessary to neutralize the alpha-synuclein protein (e.g., human alpha-synuclein protein) in the brain of the subject (e.g., the amount that is necessary to prevent or reduce the spreading of alpha-synuclein protein in the brain of the subject) is determined based on an amount of total alpha-synuclein protein in a sample, an amount of alpha-synuclein protein in a sample that is bound to the isolated antibody or multispecific antibody, and/or the amount of free alpha-synuclein protein in a sample.

[0878] In some embodiments, the amount of the isolated antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody that is administered to the subject (e.g., in order to prevent or treat a disease such as a neurodegenerative disease) is determined based on an amount of total alpha-synuclein protein in a sample, an amount of alpha-synuclein protein in a sample that is bound to the isolated antibody or multispecific antibody, and/or the amount of free alpha-synuclein protein in a sample.

[0879] In some embodiments, the sample is obtained from the subject. Suitable samples include, but are not limited to, interstitial brain fluid samples, cerebrospinal fluid (CSF) samples, blood samples, or any combination thereof. When the sample(s) are obtained from the subject, the sample(s) can be obtained before treatment and/or during treatment. In some embodiments, the amount of total, bound, and/or free alpha-synuclein antibody in the sample(s) can be used to adjust treatment (e.g., increase or decrease the amount of the isolated antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody that is administered to the subject, or to select a different isolated antibody, pharmaceutical composition, or multispecific antibody for administration to the subject).

[0880] In some embodiments, an anti-alpha-synuclein antibody that is specific for, or has a higher binding affinity for, atoxic species of alpha-synuclein (e.g., oligomeric alpha-synuclein) is used to determine the amount of bound alpha-synuclein protein. An example of measuring the amounts of bound and free alpha-synuclein protein are provided herein, e.g., in Example 6 below.

[0881] In some embodiments, an amount of the isolated antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody is administered to a subject (e.g., an animal or a human subject) such that at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%), 50%), 55%), 60%), 65%, or 70% of alpha-synuclein protein (e.g., in sample obtained from the subject, or in the brain of the subject) is bound by the isolated antibody or multispecific antibody. Preferably, an amount of the isolated antibody, pharmaceutical composition, or multispecific antibody is administered to a subject such that at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% of alpha- synuclein protein is bound by the isolated antibody or multispecific antibody. Even more preferably, an amount of the isolated antibody, pharmaceutical composition, or multispecific antibody is administered to a subject such that at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%), 98%), or 99% of alpha-synuclein protein is bound by the isolated antibody or multispecific antibody.

[0882] Typically, higher percentages of bound alpha-synuclein protein (i.e., bound by an isolated antibody or multispecific (e.g., bispecific) antibody described herein) are preferred, as a higher percentage of alpha-synuclein protein is associated with a reduction in alpha-synuclein spreading, oligomerization, or aggregation. In some embodiments, the percentage of bound alpha-synuclein protein that is necessary to inhibit alpha-synuclein spreading, oligomerization, or aggregation is used to determine how much isolated antibody, pharmaceutical composition, or multispecific antibody is administered to a subject (e.g., in order to treat the subject).

[0883] Animal models can be used to determine an appropriate amount of anti-alpha- synuclein antibody to be administered to a human subject (e.g., in order to successfully inhibit alpha-synuclein spreading, oligomerization, or aggregation). As a non-limiting example, since the levels of alpha-synuclein protein in mouse interstitial fluid (ISF), CSF, and blood (e.g., plasma) are similar to the corresponding levels of alpha-synuclein protein in humans, a mouse model can be used to determine how much anti-alpha-synuclein protein needs to be administered to a mouse in order to achieve a desired percentage of bound alpha-synuclein, a desired reduction in alpha-synuclein spreading, oligomerization, or aggregation, or a desired therapeutic effect (e.g., a reduction in signs or symptoms of a disease). The dose can then be extrapolated from the mouse model to arrive at an appropriate dose in humans.

[0884] As a non-limiting example, a transgenic animal in which the animal overexpresses alpha-synuclein can be used as model of alpha-synuclein spreading. Such models are useful for assessing the amount of an anti-alpha-synuclein antibody that is necessary to inhibit alpha- synuclein spreading and/or establishing a relationship between a particular antibody dose and the resulting percentage of bound alpha-synuclein. In some embodiments, the amount of anti- alpha-synuclein antibody that needs to be administered to a mouse in to achieve about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% bound alpha-synuclein (e.g., in an ISF, CSF, or blood sample) can be determined, and then the dose can be extrapolated to humans. In some embodiments, the amount of anti-alpha-synuclein antibody that needs to be administered to a mouse in to decrease alpha-synuclein spreading by about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% can be determined, and then the dose can be extrapolated to humans.

[0885] In some embodiments, the amount of the isolated antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody that is necessary to neutralize the alpha-synuclein protein (e.g., human alpha-synuclein protein) in the brain of the subject (e.g., the amount that is necessary to prevent or reduce the spreading of alpha-synuclein protein in the brain of the subject) is determined based on a level of disease progression (e.g., neurodegenerative disease progression) in the subject. In some embodiments, the amount of the isolated antibody, pharmaceutical composition, or multispecific antibody that is necessary to neutralize the alpha-synuclein protein in the brain of the subject is determined based on a level of alpha-synuclein protein spreading (e.g., in the brain of the subject).

[0886] In some embodiments, the amount of the isolated antibody, pharmaceutical composition, or multispecific (e.g., bispecific) antibody that is administered to the subject (e.g., in order to treat a disease such as a neurodegenerative disease) is determined based on a level of disease progression (e.g., neurodegenerative disease progression) in the subject. In some embodiments, the amount of the isolated antibody, pharmaceutical composition, or multispecific antibody that is administered to the subject is determined based on a level of alpha-synuclein protein spreading (e.g., in the brain of the subject).

[0887] Various methods can be used to assess alpha-synuclein spreading. In some embodiments, immunohistochemistry (IHC) is used. For IHC methods, an antibody that is specific for phospho-129 alpha-synuclein can be used to detect the presence or extent of spreading of alpha-synuclein (e.g., in a brain of a subject such as an animal subject).

[0888] In some embodiments, the amount of alpha-synuclein protein is determined in a sample (e.g., a CSF sample, ISF, sample, or blood sample) in order to determine disease progression in a subject. Typically, higher amounts of alpha-synuclein protein are associated with more advanced disease progression. As a non-limiting example, ELISA can be used to measure the amount of alpha-synuclein protein that is present in a sample, e.g., using antibodies that are specific for phospho-129 alpha-synuclein.

[0889] In some embodiments, an imaging method is used to determine the level of disease progression (e.g., neurodegenerative disease progression) in the subject and/or to determine the amount of alpha-synuclein protein (e.g., human alpha-synuclein protein) spreading (e.g., in the brain of the subject). In some embodiments, an imaging method is used to determine the level of disease progression in the subject. In some embodiments, an imaging method is used to determine the amount of alpha-synuclein protein spreading. In some embodiments, an imaging method is used to determine the level of disease progression in the subject and the amount of alpha-synuclein protein spreading. In some embodiments, two or more different imaging methods are used. Non-limiting examples of suitable imaging methods include positron emission tomography (PET) imaging, dopamine transporter (DaT) imaging, magnetic resonance imaging (MRI), and computed tomography (CT) imaging.

[0890] Assessing the extent of alpha-synuclein spreading and/or disease progression in a subject is not only useful for preclinical studies and clinical trials, but can also be used for patient selection. In some embodiments, the extent of alpha-synuclein spreading and/or disease progression is used to select an appropriate therapy for a subj ect and/or determine whether a particular therapy should be used or discontinued (e.g., by determining antibody activity or response to treatment). In some embodiments, the extent of alpha-synuclein spreading and/or disease progression is used to determine whether a particular subject is suitable for a clinical trial. [0891] In some embodiments, dopamine transporter (DaT) imaging can be used to determine disease progression. DaT imaging comprises administering (e.g., intravenously administering) ¹²³I-ioflupane (N-ro-fluoropropyl-2P-carbomethoxy-3P-(4-iodophenyl) nortropane), which is a single photon emission computer tomography (SPECT) molecular imaging agent, to a subject. ²³I-ioflupane is a cocaine analog that has high affinity and good selectivity for DaT. In particular neurodegenerative diseases such as Parkinson' s disease (PD), multiple system atrophy (MSA), and progressive supranuclear palsy (PSP), disease progression is associated with presynaptic dopaminergic degeneration and decreased DaT binding. In addition, ²³I- ioflupane-SPECT imaging can be used to differentiate dementia with Lewy bodies from other forms of dementia.

[0892] PET imaging can also be used to determine disease progression in a subject and/or the amount of alpha-synuclein spreading. In some embodiments, a PET probe that is specific for phospho-129 is used. In other embodiments, a diabody of an anti-alpha-synuclein antibody described herein (e.g., a diabody having a short half-life) is labeled with an appropriate tracer and used as a PET probe.

[0893] In some embodiments, the method further comprises administering to the subject one or more other therapeutic agents. In some embodiments, the method further comprises administering to the subject an agent, e.g., an antibody, that binds to amyloid beta peptides or prevents the aggregation of amyloid beta peptides. In some embodiments, the method further comprises administering to the subject an antibody against amyloid beta (Αβ), including but not limited to aducanumab, bapineuzumab, solanezumab, and gantenerumab. In some embodiments, the method further comprises administering to the subject a Beta-site Amyloid precursor protein Cleaving Enzyme 1 (BACE1) inhibitor, including but not limited to verubecestat. In some embodiments, the method further comprises administering to the subject a neuroprotective agent. In some embodiments, the neuroprotective agent is an anticholinergic agent, a dopaminergic agent, a glutamatergic agent, a histone deacetylase (HDAC) inhibitor, a cannabinoid, a caspase inhibitor, melatonin, an anti -inflammatory agent, a hormone (e.g., estrogen or progesterone), or a vitamin. In some embodiments, the method further comprises administering to the subject an agent for use in treating a cognitive or behavioral symptom of a neurodegenerative disease (e.g., an antidepressant, a dopamine agonist, or an anti-psychotic).

[0894] In some embodiments, an anti-alpha-synuclein antibody is administered to a subject at a therapeutically effective amount or dose. A daily dose range of about 0.01 mg/kg to about 500 mg/kg, or about 0.1 mg/kg to about 200 mg/kg, or about 1 mg/kg to about 100 mg/kg, or about 10 mg/kg to about 50 mg/kg, can be used. The dosages, however, may be varied according to several factors, including the chosen route of administration, the formulation of the composition, patient response, the severity of the condition, the subject's weight, and the judgment of the prescribing physician. The dosage can be increased or decreased over time, as required by an individual patient. In certain instances, a patient initially is given a low dose, which is then increased to an efficacious dosage tolerable to the patient. Determination of an effective amount is well within the capability of those skilled in the art.

[0895] The route of administration of an anti-alpha-synuclein antibody (or antigen-binding fragment, multispecific (e.g., bispecific) antibody, or pharmaceutical composition) as described herein can be oral, intraperitoneal, transdermal, subcutaneous, intravenous, intramuscular, intrathecal, inhalational, topical, intralesional, rectal, intrabronchial, nasal, transmucosal, intestinal, ocular or otic delivery, or any other methods known in the art. In some embodiments, the antibody is administered orally, intravenously, or intraperitoneally.

[0896] Co-administered agents (e.g., the anti-alpha-synuclein antibody and another therapeutic agent) can be administered together or separately, simultaneously or at different times. When administered, the therapeutic agents independently can be administered once, twice, three, four times daily or more or less often, as needed. In some embodiments, the administered therapeutic agents are administered once daily. In some embodiments, the administered therapeutic agents are administered at the same time or times, for instance as an admixture. In some embodiments, one or more of the therapeutic agents is administered in a sustained-release formulation.

[0897] In some embodiments, the anti-alpha-synuclein antibody and another therapeutic agent are administered concurrently. In some embodiments, the anti-alpha-synuclein antibody and another therapeutic agent are administered sequentially. For example, in some embodiments an anti-alpha-synuclein antibody is administered first, for example for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 days or more prior to administering another therapeutic agent. In some embodiments, the other therapeutic agent is administered first, for example, for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 days or more prior to administering an anti-alpha-synuclein antibody. [0898] In some embodiments, the anti-alpha-synuclein antibody (and optionally another therapeutic agent) is administered to the subject over an extended period of time, e.g., for at least 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350 days or longer.

IX. PHARMACEUTICAL COMPOSITIONS AND KITS

[0899] In another aspect, pharmaceutical compositions and kits comprising an antibody that specifically binds to a human alpha-synuclein protein are provided. In some embodiments, the pharmaceutical compositions and kits are for use in neutralizing human alpha-synuclein protein, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha- synuclein protein comprises preventing, reducing, or inhibiting alpha-synuclein oligomerization and/or aggregation in a subject, e.g., in a brain of a subject. In some embodiments, neutralizing the human alpha-synuclein protein comprises preventing, reducing, or inhibiting the spreading of alpha-synuclein protein oligomers and/or fibrils in a subject, e.g., in a brain of a subject. In some embodiments, the pharmaceutical compositions and kits are for use in treating a neurodegenerative disease (e.g., a synucleinopathy or tauopathy).

Pharmaceutical compositions

[0900] In some embodiments, pharmaceutical compositions comprising an anti-alpha- synuclein antibody are provided. In some embodiments, the anti-alpha-synuclein antibody is an antibody (or antigen-binding fragment or multispecific (e.g., bispecific) antibody) as described in Section III above.

[0901] In some embodiments, a pharmaceutical composition comprises an anti-alpha- synuclein antibody as described herein and further comprises one or more pharmaceutically acceptable carriers and/or excipients. A pharmaceutically acceptable carrier includes any solvents, dispersion media, or coatings that are physiologically compatible and that preferably does not interfere with or otherwise inhibit the activity of the active agent. Various pharmaceutically acceptable excipients are well-known in the art.

[0902] In some embodiments, the carrier is suitable for intravenous, intramuscular, oral, intraperitoneal, intrathecal, transdermal, topical, or subcutaneous administration. Pharmaceutically acceptable carriers can contain one or more physiologically acceptable compound(s) that act, for example, to stabilize the composition or to increase or decrease the absorption of the active agent(s). Physiologically acceptable compounds can include, for example, carbohydrates, such as glucose, sucrose, or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, compositions that reduce the clearance or hydrolysis of the active agents, or excipients or other stabilizers and/or buffers. Other pharmaceutically acceptable carriers and their formulations are well-known in the art.

[0903] The pharmaceutical compositions described herein can be manufactured in a manner that is known to those of skill in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating, entrapping or lyophilizing processes. The following methods and excipients are merely exemplary and are in no way limiting.

[0904] For oral administration, an anti-alpha-synuclein antibody can be formulated by combining it with pharmaceutically acceptable carriers that are well known in the art. Such carriers enable the compounds to be formulated as tablets, pills, dragees, capsules, emulsions, lipophilic and hydrophilic suspensions, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by mixing the compounds with a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, such as a cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0905] An anti-alpha-synuclein antibody can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. For injection, the compound or compounds can be formulated into preparations by dissolving, suspending or emulsifying them in an aqueous or nonaqueous solvent, such as vegetable or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives. In some embodiments, compounds can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' s solution, Ringer's solution, or physiological saline buffer. Formulations for injection can be presented in unit dosage form, e.g., in ampules or in multi-dose containers, with an added preservative. The compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

[0906] In some embodiments, an anti-alpha-synuclein antibody is prepared for delivery in a sustained-release, controlled release, extended-release, timed-release or delayed-release formulation, for example, in semi-permeable matrices of solid hydrophobic polymers containing the active agent. Various types of sustained-release materials have been established and are well known by those skilled in the art. Current extended-release formulations include film-coated tablets, multiparticulate or pellet systems, matrix technologies using hydrophilic or lipophilic materials and wax-based tablets with pore-forming excipients. Sustained-release delivery systems can, depending on their design, release the compounds over the course of hours or days, for instance, over 4, 6, 8, 10, 12, 16, 20, 24 hours or more. Usually, sustained release formulations can be prepared using naturally occurring or synthetic polymers, for instance, polymeric vinyl pyrrolidones, such as polyvinyl pyrrolidone (PVP); carboxyvinyl hydrophilic polymers; hydrophobic and/or hydrophilic hydrocolloids, such as methylcellulose, ethylcellulose, hydroxypropylcellulose, and hydroxypropylmethylcellulose; and carboxypolymethylene.

[0907] Typically, a pharmaceutical composition for use in in vivo administration is sterile. Sterilization can be accomplished according to methods known in the art, e.g., heat sterilization, steam sterilization, sterile filtration, or irradiation.

[0908] Dosages and desired drug concentration of pharmaceutical compositions of the disclosure may vary depending on the particular use envisioned. The determination of the appropriate dosage or route of administration is well within the skill of one in the art. Suitable dosages are also described in Section IV above.

Kits

[0909] In some embodiments, kits comprising an anti-alpha-synuclein antibody (or antigen- binding fragment or multispecific {e.g., bispecific) antibody) as described herein {e.g., as described in Section III above) are provided.

[0910] In some embodiments, the kit further comprises one or more additional therapeutic agents. For example, in some embodiments, the kit comprises an anti-alpha-synuclein antibody as described herein and further comprises one or more additional therapeutic agents for use in the treatment of a neurodegenerative disease. In some embodiments, the therapeutic agent is an agent for use in treating a cognitive or behavioral symptom of a neurodegenerative disease (e.g., an antidepressant, a dopamine agonist, or an anti-psychotic). In some embodiments, the therapeutic agent is a neuroprotective agent (e.g., an anticholinergic agent, a dopaminergic agent, a glutamatergic agent, a histone deacetylase (HDAC) inhibitor, a cannabinoid, a caspase inhibitor, melatonin, an anti-inflammatory agent, a hormone (e.g., estrogen or progesterone), or a vitamin).

[0911] In some embodiments, the kit further comprises instructional materials containing directions (i.e., protocols) for the practice of the methods described herein (e.g., instructions for using the kit for treating a neurodegenerative disease (e.g., a synucleinopathy or tauopathy)). While the instructional materials typically comprise written or printed materials they are not limited to such. Any medium capable of storing such instructions and communicating them to an end user is contemplated by this invention. Such media include, but are not limited to electronic storage media (e.g., magnetic discs, tapes, cartridges, chips), optical media (e.g., CD-ROM), and the like. Such media may include addresses to internet sites that provide such instructional materials.

X. EXAMPLES

[0912] The present invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes only, and are not intended to limit the invention in any manner.

Example 1. Preparation of an alpha-synuclein immunogen composition for anti-alpha- synuclein antibody discovery and screening

[0913] This example describes the design and preparation of an immunogenic composition for the discovery of alpha-synuclein antibodies. Alpha-synuclein is suspected to play a central role in neuronal cell death during neurodegenerative diseases, such as Parkinson' s disease. Alpha-synuclein exists in multiple specific forms, including species of monomer, truncated monomers, and various oligomers, and fibrils of monomers and truncated monomers. The monomelic species can be less immunogenic compared to oligomers and fibrils. The oligomer species, is more acidic, immunogenic and the C-terminal region is highly dynamic in the oligomeric state, thus providing a unique confirmation to generate alpha-synuclein antibodies that bind multiple species. [0914] Human alpha-synuclein protein (Anaspec AS55555) was dissolved at a concentration of 1 mg/mL in PBS and verified to be >95% monomeric by size exclusion chromatography. Alpha-synuclein was oligomerized according to Fink and coworkers, J. Biol Chem, 2007. Briefly, alpha-synuclein was concentrated to 2 mg/mL in PBS and mixed with a solution of 4- hydroxy-nonenal (4HNE, Cayman, 10 mg/mL in ethanol), at a final molar ratio of 1 :20 protein/4HNE and incubated overnight at 37 °C. The resulting mixture was analyzed and found to consist of approximately 80% oligomers by area under the curve on analytical size exclusion chromatography, with the remainder of alpha-synuclein being monomeric species in equilibrium. The composition was used without further purification as an immunogenic composition.

Example 2. Anti-alpha-synuclein antibody discovery and screening

[0915] This example describes the design, generation, and characterization of antibodies that specifically bind to monomers and oligomers of human alpha-synuclein protein. The antibodies also exhibit cross-species reactivity, in some instances, between human alpha- synuclein and mouse alpha-synuclein.

Preparation of biotinylated alpha-synuclein monomer and oligomers

[0916] Human alpha-synuclein protein (Anaspec AS55555) was dissolved at a concentration of 1 mg/mL in PBS and verified to be >95% monomeric by size exclusion chromatography. An aliquot of monomeric alpha-synuclein was biotinylated using EZ-link Sulfo- HS biotin (Pierce) according to the manufacturer's instructions and de-salted using an Amicon ultra spin- concentrator (3,000 molecular weight cut-off). Extent of biotinylation was determined through intact-mass determination using a Thermo Exactive EMR. Each alpha-synuclein molecule was biotinylated at a molar ratio of approximately 3-7 molecules of biotin/molecule of alpha- synuclein. Alpha-synuclein was oligomerized according to Fink and co-workers, J. Biol Chem, 2007. Briefly, biotinylated alpha-synuclein was concentrated to 2 mg/mL in PBS and mixed with a solution of 4-hydroxynonenal (4HNE, Cayman, 10 mg/mL in ethanol), at a final molar ratio of 1 :20 protein/4HNE and incubated overnight at 37 °C. Oligomers were purified from remaining monomeric biotinylated alpha-synuclein by size-exclusion chromatography and snap-frozen and stored at -80 °C until use. Immunization of mice

[0917] Alpha-synuclein knockout mice (obtained from Jackson Laboratory) were immunized with recombinant alpha-synuclein, either monomers or oligomers. Immunizations were performed via Hock or footpad weekly with 5-10 μg of antigen in Ribi adjuvant until serum antibody reacted against alpha-synuclein protein in an ELISA to a dilution of >10⁵ (4-6 weeks). Mice were given a final boost of 5μg of antigen without adjuvant via intraperitoneal injection and sacrificed three days later. Popliteal and inguinal lymph nodes were harvested, made into single-cell suspensions by passing through cell strainers, and finally flash frozen in liquid nitrogen.

Purification of RNA and generation of cDNA from splenocytes and lymphocytes

[0918] Cell pellets from frozen lymphocytes were thawed on ice and resuspended in 1 mL TRIzol^™ The solution was incubated at room temperature for 5 minutes, then 0.2 mL of chloroform was added and the tubes were shaken vigorously for 15 seconds. The tubes were spun at 20 x g for 15 minutes at 4 °C to separate the phases. The colorless aqueous phase was carefully removed, leaving the white interphase and the pink chloroform phase. RNA was precipitated by addition of 0.6 mL of isopropanol, mixing by inversion, and then spinning at 20 x g for 15 minutes at 4 °C. The supernatant was discarded and the pellet was washed with 75% ethanol and dissolved in DEPC-treated water. cDNA was generated from the total RNA using Superscript^™ III reverse transcriptase (obtained from Thermo Fisher) using the manufacturer's recommended protocol for priming with oligo-dT.

Generation of immunized phage display libraries

[0919] PCR reactions were carried out to separately amplify V genes for heavy and light chain variable regions using primers specific for the respective mouse germline genes. The PCR products were purified by agarose gel. VH-CH1 fragments were generated by a joining PCR reaction using the VH products and DNA for human CHI region along with end primers. VL-CL fragments were generated by a joining PCR reaction using the VL products and DNA for the human kappa region along with end primers. The respective fragments were purified by agarose gel. A final PCR reaction was carried out to join the VH-CHl and VL-CL fragments, and the resulting DNA encoding Fab was again purified by agarose gel, then digested with Sfil restriction enzyme. The digested Fab fragment was ligated into the Sfil- digested phagemid vector overnight at 16 °C, and the ligation product was purified by ethanol precipitation. Electrocompetent TGI E. coli cells (obtained from Lucigen^®) were electroporated using a BTX^® ECM^® 630 electroporation system and the manufacturer's suggested parameters. For each library, eight to ten electroporation reactions with 0.5 μg of ligation product per reaction were performed. Immediately after each electroporation reaction, the cuvettes were washed with 2 mL of recovery media (obtained from Lucigen^®) and the TGI cells were recovered at 37 °C with shaking for 1 hour. A small amount of bacteria were saved at this point and dilution-plated to determine the number of library transformants (~lxl0¹⁰ cfu). Selective media, 2YT containing 100 μg/mL carbenicillin was added to the recovered E. coli, and the TGI cells were grown to an OD₆oo of approximately 0.5 (mid-log growth phase). M13K07 helper phage were added at an MOI of approximately 10 and infected at 37 °C for 30 minutes without shaking, followed by 30 minutes with shaking, and then 50 μg/mL kanamycin was added. The cells were grown overnight at 30 °C with shaking. Cultures were harvested by centrifugation at 8,000 rpm at a temperature of 4 °C and the pellets were discarded. PEG/NaCl (obtained from Teknova) was added to the supernatants at a final concentration of 4% PEG. The phage were precipitated on ice for 1 hour, then spun at 8,000 rpm to pellet. The supernatants were removed, and the phage were dissolved in 40 mL PBS. The solutions were centrifuged at maximum speed for 10 minutes to remove any insoluble material, then phage were precipitated a second time from the supernatant by addition of PEG/NaCl and incubation on ice. The phage were again pelleted by centrifugation and suspended in 8 mL of PBS containing 15% glycerol, then aliquoted and frozen. Titering of phage yielded approximately 5xl0¹² cfu/mL phage concentration.

Phage panning

[0920] Phage aliquots (0.5 mL) were thawed and mixed with 0.5 mL of 10% BSA in PBS to block. The phage were added to an aliquot of 200 μΕ M280-streptavidin (M280SA) beads and incubated at room temperature with gentle rotation for 1 hour. Separate campaigns were performed with either biotinylated monomelic alpha-synuclein or biotinylated oligomeric alpha-synuclein, both prepared as described herein. Meanwhile, biotinylated alpha-synuclein (100 pmol) was added to a second aliquot of 200 μΕ M280SA beads and incubated at room temperature with gentle rotation for 30 minutes, then the beads were washed with PBS with 1%) BSA (PBS A) three times, using a magnet to separate the beads between each wash. The phage solution was then separated from the negative M280SA beads using the magnet, and added to the alpha-synuclein-loaded beads. After a one-hour incubation with gentle rotation, the beads were washed with PBS containing 0.1% BSA and 0.05% Tween^® 20 (4x total, two washes for 5 minutes). Bound phage were eluted from the beads using 0.1 M glycine (pH 2.7) for 30 minutes; the eluted phage solution was then neutralized with 1 M Tris (pH 7.5). The eluted phage were used to infect 10 mL TGI E. coli grown to mid-log phase (OD₆oo approximately 0.5) at 37 °C for 30 minutes without shaking, followed by 30 minutes with shaking. Selective media (2YTC) was added (40 mL volume), and the cultures were grown for 1-2 hours at 37 °C with shaking. At this point the cultures were at OD₆oo of 0.5 or lower, and were infected with M13K07 helper phage at an MOI of approximately 10. After infection, 50 μg/mL kanamycin was added and the cultures were grown overnight at 30 °C with shaking.

[0921] For the second panning round, phage were precipitated once with PEG/NaCl and concentrated 20-fold, then the same general protocol as above was followed (identical conditions for panning on either monomers or oligomers). For the washing step, the beads from each condition were split into two pools. One pool was subjected to standard washing conditions: 6x PBST washes, 2 for 10 minutes). The second pool was subjected to more stringent washing conditions: 8x PBST washes, 2 for 60 minutes. After the second panning round, infected TGI cells from each pool were spread onto 2YTCG plates (obtained from Teknova) for picking of single colonies for screening.

Generating Fabs via periplasmic expression

[0922] Single colonies from the second panning round output were picked and placed into 96-well deep-well plates containing 0.5 mL of 2YTC media (a total of 1,692 colonies). The plates were sealed and grown overnight at 37 °C with shaking. To induce periplasmic Fab expression, 50 iL of the overnight culture was transferred to new 96-well deep-well plates containing 950 iL per well of 2YTC media with auto-induction supplements (obtained from EMD Millipore). The plates were sealed and grown overnight at 37 °C. The plates were centrifuged at 4,000 rpm for 10 minutes to pellet the bacteria and the media was discarded. The pellets were suspended in 0.1 mL of PPB buffer (obtained from Teknova) and shaken at 10 °C and 1,000 rpm for one hour, after which 0.3 mL of water was added to each well. After an additional 30 minutes of shaking, the plates were spun at maximum speed for 10 minutes. The resulting supernatant contained soluble, crude Fab for screening.

Screening Fabs for binding to alpha-synuclein (FAB expressed on phage)

[0923] 384-well ELISA plates were coated overnight with 1 μg/mL streptavidin for capturing biotinylated antigen. The plates were washed 3x with PBST and blocked with PBS+1% BSA (PBSA). After one hour of blocking, biotin-alpha-synuclein monomer (0.5 μg/mL), biotin-alpha-synuclein oligomer (0.5 μg/mL), or block was added to the streptavidin wells. After washing wells 3x with PBST, each Fab periplasmic extract was subsequently added to wells containing either streptavidin alone, biotin-alpha-synuclein monomer, or biotin- alpha-synuclein oligomer. The plates were incubated at room temperature for one hour, then washed 3x with PBST. Secondary detection antibody, anti-kappa-HRP (obtained from Bethyl Laboratories), was added at 1 :20,000 in PBSA, and incubated at room temperature for 20 minutes. The plates were washed 3x with PBST and developed with TMB substrate (Thermo Fisher) and quenched with 2 N sulfuric acid. The signal was quantified on a BioTek® plate reader at A450. Wells with signals greater than 5-fold over background against alpha-synuclein monomer and/or oligomer, also with less than 2-fold over background against streptavidin, were considered positive (around 60-70% of clones). Bacteria colonies from the respective positive wells were submitted for rolling-circle-amplification and sequencing of the variable genes. The sequenced clones could be sorted into approximately 28 different gene family lineages (e.g., groups with closely related CDRs in terms of length and sequence).

[0924] Clones with unique sequences (241 total) were re-arrayed to new plates, and a similar ELISA was subsequently performed to determine whether any of the antibodies bound to fragments of alpha-synuclein using directly coated polypeptides of alpha-synuclein corresponding to sequence positions 1-60, 1 -95, 1-1 12, 61-140, 96-140, 51-67, and 96-1 10. Testing was also performed to determine whether any of the antibodies bound to beta-synuclein or gamma-synuclein in an analogous single-point ELISA. Results for unique hit sequences are found in Table 1.

Reformatting and expression of chimeric or murine IgGs

[0925] It was observed that most sequence families appeared in clones panned against both monomeric and oligomeric alpha-synuclein, supporting the screening results showing cross- reactivity to both alpha-synuclein forms. A panel of antibodies was selected based on the screening and sequence data for reformatting recombinant expression as human IgGl Fc polypeptides. Criteria for selection was to fully encompass all sequence families; for more diverse sequence families, at least 2-4 different antibodies were selected, based on those with the most favorable binding properties during screening or the greatest sequence differences. The heavy and light chain variable region sequences of these hits were cloned into mammalian expression vectors containing human IgGl and kappa constant regions, respectively. The antibodies were expressed in 293F cells and purified by Protein A chromatography using standard methods that will be known to one of ordinary skill in the art.

Engineering 1F06 antibody

[0926] Antibody 1F06 was found to have a cysteine at position 99 (numbered using the Kabat system) in CDR-H3. Since cysteines can cause development problems, it was explored whether this CDR residue could be mutated without compromising the high affinity of this antibody. Variants were made wherein the cysteine was replaced by either serine, alanine, or methionine. Testing of these variants by Biacore™, as described below, showed that the C99M variant had similar affinity and binding to alpha-synuclein as compared to 1F06, whereas C99A and C99S had significantly reduced binding (numbered using the Kabat system).

Biacore™ kinetics

[0927] Anti-human Fc antibody (obtained from GE Healthcare) was immobilized on the surface of a CM5 chip (obtained from GE Healthcare) through amine-coupling to reach about 6,000 to 8,000 response units (RU). The surface was activated by injection of a mixture of 1- ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS), both obtained from GE Healthcare, for 7 minutes. Anti-human Fc antibody was diluted in sodium acetate (pH 5.0) at 25 μg/mL and injected for 10 minutes at a flow rate of 5 μΕ/πήηυΐε, followed by injection of ethanolamine (obtained from GE Healthcare) for 7 minutes.

[0928] Purified anti alpha-synuclein antibody (10 μg/mL) was captured to reach 1,500 to 2,000 RU. A range of serially-diluted alpha-synuclein monomer (e.g., 3.4 nM to 300 nM) was injected at a flow rate of 30 μΐνπήηυΐε, using either single-cycle kinetics or full-cycle kinetics methods. Sensorgrams were fitted using a 1 : 1 Langmuir model to estimate k₀n and k₀ff. Table 2 provides kinetic parameters for antibodies tested using single-cycle kinetics, and Table 3 provides kinetic parameters for antibodies tested using full-cycle-kinetics. These data show that the selected antibodies had affinities ranging from around 0.92-33 nM. Notably, many of the single-digit nM affinity antibodies had very slow off-rates with k₀ff values less than 5.0x10^" ⁴ s^"1. It is also noted that the C99M variant of 1F06 had a similar or slightly better kinetics profile as compared to the parental 1F06 antibody based on single-cycle kinetics.

[0929] Kinetics for binding to alpha-synuclein oligomers, prepared as described above for immunization, were determined using the full-cycle kinetics method as described for the monomers. It is important to note that the solution concentration of oligomers was estimated based on the extinction coefficient of monomers and therefore may not accurately reflect the number of oligomeric molecules in solution. It is also important to note that the k₀n and KD values may vary with different preparation lots of oligomer; conversely the k₀ff value is not dependent on antigen concentration and should therefore remain consistent. Table 4 shows that the selected antibodies had k₀ff values for alpha-synuclein oligomers ranging from around 6.0xl0^"4 to 1.3xl0^"3 s^"1, and affinities for alpha-synuclein oligomers ranging from around 2.5- 38 nM.

ELISA binding to alpha-synuclein

[0930] Half-area 96-well ELISA plates were coated with various synucleins (i.e., monomeric human alpha-synuclein, oligomeric human alpha-synuclein, human beta-synuclein, human gamma-synuclein, and mouse alpha-synuclein) at 0.5 μg/mL overnight at 4 °C. The plates were washed 3x with PBST and blocked with PBS A for one hour at room temperature. The chimeric anti-alpha-synuclein IgGs were added across the plate at 3 -fold serial dilutions starting at 200 nM, and the plates were incubated for 1 hour at room temperature. After 3x washes with PBST, anti-kappa-HRP antibody was added at 1 :5,000 dilution and the plates were incubated at room temperature for 20 minutes. The plates were washed again 3x with PBST and developed with TMB substrate which was quenched with 2 N sulfuric acid. The A450 values were determined using a BioTek^® plate reader, and data was processed using Prism software. EC50 data is presented in Table 5. These data show that the antibodies tested are highly specific for alpha-synuclein over beta- and gamma-synuclein, and that most bind monomeric and oligomeric alpha-synuclein similarly. Furthermore, most of the antibodies strongly cross-react with mouse alpha-synuclein.

Epitope mapping

[0931] A peptide microarray spotted with synthetic 15-mers with an overlap of 10 amino acids (JPT, Berlin, Germany) covering the full-length human alpha-synuclein (SEQ ID NO: l; P 000336.1) was used for epitope mapping. Spots of human IgG and mouse IgG were included as positive controls. Monoclonal antibodies (either mouse variable-human IgGl chimeras, or mouse variable-mouse IgG2a) diluted to a concentration of 10 μg/mL in TBST with 3% BSA were incubated on glass slides for 2 hours at room temperature. After washing 5x with TBST, Alexa Fluor^® 568 coupled to goat anti-human IgG (H+L) or Alexa Fluor^® 647 coupled to goat anti -mouse IgG2b (obtained from Life technologies; 1 μg/mL in TBST with 3% BSA) was added for 1 hour protected from light. The slides were washed 5x with TBST and 5x with deionized water to remove excess salts, then dried under nitrogen. The slides were imaged with an Opera Phenix^™ slide scanner (obtained from Perkin Elmer^®, Waltham, MA) and analyzed using ImageJ/TrackEM software version 2.0.0. The sequences of 15-mers that bound to one or more clones are listed in Table 6. Epitopes for clones 1F06, 2D02, 2F09, 2F10, and 3C02 are listed in Table 7.

Pharmacokinetic studies

[0932] Wild-type 6-8 week old male C57B16 mice (n=3/group) are intravenously dosed with 10 mg/kg of antibody (e.g., 1F06, 2D02, 2F09, 2F10, or 3C02). In-life bleeds are taken at 30 minutes, 4 hours, 1 day, 3 days, 7 days, 10 days, and 14 days post-dose via submandibular bleeds. Blood is collected in K2EDTA plasma tubes and spun at 2000g for 10 minutes at 4 °C. Plasma is then isolated for subsequent pharmacokinetic analysis. A custom anti-mouse IgG assay is developed to quantify mouse anti-alpha-synuclein antibodies in plasma. First, alpha- synuclein protein (1 μg/mL) is immobilized on a plate. Plasma diluted to 1 :2,500 is then incubated with the bound alpha-synuclein protein followed by an HRP (horse radish- peroxidase) conjugated polyclonal antibody. A TMB (tetra methyl benzidine) substrate is used for the color development and its signal is directly proportional to the concentration of antibody bound to the alpha-synuclein protein. Dosing solutions are also analyzed on the same plate to confirm the correct dosage. An independent standard curve of each mouse anti-alpha-synuclein antibody is used (1 μg/mL to 0.0004 ug/mL) to quantify the IgG in plasma and a curve fit is done using a four-parameter logistic regression. This example demonstrates that antibodies of the present invention exhibit certain in vivo pharmacokinetic properties.

Immunodepletion of alpha-synuclein in Parkinson's disease patient CSF

[0933] Cerebrospinal fluid (CSF) previously obtained from Parkinson' s disease patients was obtained from PrecisionMed. Magnetic Protein A beads (SureBeads; Bio-Rad) were first blocked with 2% BSA in TBS for 1 hour at 24 °C and then washed three times in TBS-t. Prior to incubation with antibodies, Tween-20 (0.02%) and EDTA-free complete protease inhibitors (Roche) were added to CSF. 2 μg of antibody was added to 50 μΙ_, of CSF and incubated for 1 hour at 4 °C. 100 μΙ_, of blocked beads were then added to the CSF/antibody mixture and incubated for 18 hours at 4 °C. The unbound fraction was then collected and measured using a U-PLEX human alpha-synuclein Kit (MSD #K151WKK) according to the manufacturer's instructions. All alpha-synuclein antibodies were capable of immuno-depleting some alpha- synuclein from Parkinson's disease patient CSF compared to a negative control, and antibodies 2D02 and 2F 10 removed all detectable alpha-synuclein (FIG. 1). Given the high levels of antibody that were used in this experiment, this data indicates that there was a difference in the availability of the epitopes or species of alpha-synuclein bound by these antibodies, and is not a reflection of antibody affinity (which is known to be very similar between 2F09 and 2F 10).

Example 3. Efficacy study to measure reduction of alpha-synuclein levels in the interstitial fluid (ISF) of mice following antibody administration

[0934] This example demonstrates an efficacy study using the push-pull microdialysis method that was performed using wild-type mice to measure reduction of alpha-synuclein levels in the ISF following administration of an antibody described herein.

[0935] A unilateral guide cannula was stereotaxically implanted just above the striatum of wild-type mice expressing wild-type alpha-synuclein (obtained from Jackson Laboratories; N=30). Alternatively, transgenic mice overexpressing wild-type alpha-synuclein (available, for example, from Jackson Laboratories) can be used. After implantation of the guide cannula, mice were allowed to recover for at least five days before the insertion of the microdialysis probe (2 mm, 1,000 kDa cut off probe), marking time zero. An ISF sample was collected every two hours at a constant flow rate of 0.75 μL/minute. Alternatively a constant flow rate of 1 μL/minute can be used.

[0936] Baseline ISF was collected for the first 24 hours following microdialysis probe insertion. At the 24-hour time point, animals received either a single intraperitoneal (IP) injection of control IgG (e.g., anti-IgG2; 75 mg/kg; N=10), a single intraperitoneal (IP) injection of a benchmark antibody (e.g., GM37; 75 mg/kg; N=10), or a single IP injection of an antibody according to the present invention described herein (e.g., 1F06, 2D02, 2F09, 2F10, or 3C02; 75 mg/kg; N=10). Alternatively, at the 6-hour time point, animals can receive either a single intravenous (IV) injection of control IgG (e.g., anti-RSV; 15 mg/kg; N=10), a single intravenous (IV) injection of a benchmark antibody (e.g., GM37; 15 mg/kg; N=10), or a single IV injection of an antibody according to the present invention described herein (e.g., 1F06, 2D02, 2F09, 2F 10, or 3C02; 15 mg/kg; N=10). ISF samples were collected for an additional 24 hours post injection. Blood samples were also collected before microdialysis probe insertion and at the 12-hour time point. At the termination of the experiment, animals were taken down and the cerebellum, CSF, plasma, and left hemisphere of the brain were collected for analysis. The right hemisphere of the brain was immersed in 4% PFA for histological analysis to confirm placement of the guide cannula and microdialysis probe. [0937] FIG. 2 depicts an assay that was used to measure the amounts of bound and free alpha- synuclein protein. 2F09 human IgG (2 μg/mL) was used as the coating antibody. For the bound assay, 2D02 and 2F10 (2 μg/mL) were used as detection antibodies (incubation for 15 minutes). For the free assay, biotinylated 2D02 or biotinylated 2F10 (2 μg/mL) was used as the detection antibody (incubation for 15 minutes). Results are shown in FIGS. 3 A and 3B.

[0938] FIG. 4 shows the results of assays that demonstrated a decrease in bound alpha- synuclein and a corresponding increase in free alpha-synuclein in wild-type mouse ISF. For these assays, wild-type mouse ISF was mixed with varying amounts of therapeutic antibodies (e.g., 2D02 or 2F10) and samples were incubated for 1.5 hours at room temperature with shaking. ISF was diluted 2-fold and incubated on a 384w plate with capture antibody overnight. The following day, ELISA was performed.

[0939] For the bound assay, anti-mouse HRP was used. In some embodiments, a 1-hour incubation for the detection antibody is used in the bound assay, and a dilution of 1 :5,000 for the anti-mouse HRP antibody is used, with an incubation time of 30 minutes. For the free assay, biotinylated versions of the therapeutic antibodies (2 μg/mL) were incubated for 15 minutes, followed by SA-URP (1 : 10,000) incubation for 30 minutes. In some embodiments, the standard curve can be optimized by reducing the biotinylated detection antibody concentration to 1 μg/mL (15-minute incubation time), the anti-SA HRP concentration can be reduced to 1 :20,000 to reduce background, and the final wash step can be increased from three to four washes. Controls using artificial CSF only showed no signal at all antibody concentrations, indicating that the signal was specific for alpha-synuclein.

[0940] This example demonstrates that alpha-synuclein levels in the ISF can be measured in vivo, and that the extent of alpha-synuclein reduction in ISF over time can be measured after administration of anti-alpha-synuclein antibodies of the present invention. This example also can be used to determine the extent to which the anti-alpha-synuclein antibodies of the present invention can bind soluble alpha-synuclein, including in the ISF and CNS. Reducing the levels of alpha-synuclein in the ISF over time with the antibodies of the present invention can result in alpha-synuclein neutralization in the brain and CNS, thus providing a therapeutic benefit of the anti-alpha-synuclein antibodies of the present invention, namely in the treatment of patients having diseases driven by pathological species of alpha-synuclein {e.g., Parkinson's disease). Example 4. Efficacy study to measure pathological alpha-synuclein transmission and spreading in nontransgenic or transgenic mice following antibody administration

[0941] This example demonstrates an efficacy study that can be performed using wild-type or transgenic mice overexpressing wild-type alpha-synuclein (e.g., B6/C3H Tg M83 heterozygous line) (available from Jackson Laboratories)) to measure pathological alpha- synuclein transmission or the reduction of transmission in mice brain following administration of an antibody. This example can be modeled on in vivo experiments described in Luk, K. C, et al, J. Exp. Med. 209 (5): 975 (2012), Luk, K. C, et al, Science 338(6109):949-953 (2012), or Tran, H. T., et al, Cell Rep. 7(6): 2054-2065 (2014).

[0942] Mouse pre-formed fibrils (Volpicelli-Daley, L A. et al, Nat. Protoc. 9(9):2135-46 (2014)) are unilaterally injected into the cortex and striatum of adult mice (N=30). Mice are allowed to recover and after 7 days post injection, mice receive either a single intraperitoneal (IP) injection of control IgG {e.g., anti-RSV; 50 mg/kg; N=10), a single IP injection of another benchmark antibody {e.g., GM37; 50 mg/kg; N=10), or a single IP injection of an antibody according to the present invention described herein (e.g., 1F06, 2D02, 2F09, 2F10, or 3C02; 50 mg/kg; N=10). Mice receive weekly IP injections of antibody for up to 3 months, after which the animals are sacrificed and brain tissue {e.g., at the striatum, cortex, or other sections) is studied using IHC. Pathologic alpha-synuclein is visible as phospho-alpha-synuclein {i.e., at Serl29), using a rabbit monoclonal antibody against pSerl29 (ref ab51253; Abeam) or a rabbit polyclonal antibody (pAb) against pSerl29 (ref ab59264; Abeam). The extent of alpha- synuclein transmission and spreading is measured from the striatum and cerebral cortex to the olfactory bulb, amygdala, contralateral hemisphere, substantia nigra, or other brain region. Specific neuronal pathology is measured as positive intraneuronal and neuritic inclusions in neurons, or glial pathology is alternatively measured in astrocytes.

[0943] This example demonstrates that levels of pathological alpha-synuclein in the brain of mice can be measured, and that the extent of alpha-synuclein transmission or spreading over time can be measured after administration of anti-alpha-synuclein antibodies of the present invention. This example also can be used to determine the extent to which the anti-alpha- synuclein antibodies of the present invention can bind soluble alpha-synuclein and alpha- synuclein protein fibrils, including in the brain and CNS. Reducing the levels of alpha- synuclein spreading over time with the antibodies of the present invention can result in alpha- synuclein neutralization in the brain and CNS, and thus provide a therapeutic benefit of the anti-alpha-synuclein antibodies of the present invention, namely in the treatment of patients having diseases driven by pathological species of alpha-synuclein (e.g., Parkinson's disease).

Example 5. Generation and screening of anti-alpha-synuclein antibody-producing hybridomas

[0944] This example describes the generation and screening of hybridoma cell lines that secrete anti-alpha-synuclein antibodies. This example also describes the characterization of the anti-alpha-synuclein antibodies secreted by the hybridoma cell lines. In some instances, the antibodies bind to oligomers of human alpha-synuclein with a higher affinity than that of human alpha-synuclein monomers. The antibodies also exhibit cross-species reactivity, in some instances, between human alpha-synuclein and mouse alpha-synuclein.

Immunization of mice

[0945] Alpha-synuclein knockout mice (obtained from Jackson Laboratory) were immunized with recombinant alpha-synuclein, either monomers or oligomers. Immunizations were performed via Hock or footpad weekly with 5-10 μg of antigen in Ribi adjuvant. After 6-8 injections the serum was screened for titer by protein ELISA. Once the titers were >10⁵ mice were selected for a final boost. Mice were given a final boost of 5μg of antigen without adjuvant via footpad and sacrificed three days later. Popliteal and inguinal lymph nodes were harvested, made into single-cell suspensions by passing through cell strainers, and used for fusion.

Generation of hybridoma library

[0946] B cells harvested from lymph nodes were processed and counted, then mixed with P3X63Ag8 cells at a ratio of 1 : 1 and fused using a BTX Hybrimune Electrofusion apparatus. The fused hybridomas were plated in 60-96 well plates with 100 μΐ ννεΐΐ of hypoxanthine- aminopterin-thymidine (HAT) selection media. The plates were fed with hypoxanthine thymidine (HT) after a week. After two weeks 50 μΕΛνεΙΙ of supernatant was collected and screened for antigen-specific binding.

Screening of hybridoma supernatants by ELISA

[0947] For primary screening, 96-well Nunc plates were coated with 1 μg/mL of monomeric alpha-synuclein in PBS 50 μΐ ννεΐΐ overnight. The next day, the plates were washed three times with PBST and 50 μΐ ννεΐΐ of hybridoma supernatants were transferred from fusion plates to screening plates. The plates were incubated for 1 hour at room temperature. The plates were washed three times with PBST. Secondary detection antibody goat X mouse HRP (obtained from Southern Biotech) was added 50 μΙ,ΛνεΙΙ 1 :2000 diluted in PBS. Plates were incubated for one hour at room temperature. After one hour the plates were washed three times with PBST. Plates were developed with 50 μΐ ννεΐΐ of TMB substrate (obtained from Thermo Fisher) and quenched with 50 μΐ ννεΐΐ of 1 N sulfuric acid. The signal was quantified on a BioTek^® plate reader at A450. Wells with an OD three times the background were considered positive and carried forward for secondary screening.

[0948] All the positives (i.e., about 700 clones) from the primary screen were carried forward and screened against human alpha-synuclein protein at two dilutions (i.e., neat and 1 :5). The strong binders in this assay were further screened using a range of titrations (i.e., neat, 1 : 10, 1 : 100, and 1 : 1000) against human alpha-synuclein. The clones that showed binding across the range of titrations were carried forward to be sub- cloned in semi-solid media and picked by Clonepix. The monoclonal clones were screened using ELISA in a similar manner as mentioned above.

Hybridoma sequencing protocol

[0949] Total RNA was extracted from approximately 5x 10⁶ hybridoma cells (i.e., that produced each antibody) using a Qiagen RNeasy Mini Kit. The first-strand cDNA was synthesized using a SMART RACE cDNA Amplification Kit (obtained from BD Biosciences Clontech) following the supplier's protocol. The variable region cDNAs for the heavy (VH) and light (VL) chains were amplified by polymerase chain reaction (PCR) using 3' primers that annealed, respectively, to the mouse gamma and kappa chain C regions (SEQ ID NOs:324-328 listed below), and a 5' universal primer that was provided in the SMART RACE cDNA Amplification Kit.

[0950] For VH PCR, the 3' primers were as follows: muIgGl GGACAGGGATCCAGAGTTCC (SEQ ID NO:324), muIgG2 AGCTGGGAAGGTGTGCACAC (SEQ ID NO:325), and muIgG3 CAGGGGCCAGTGGATAGAC (SEQ ID NO:326). For VL PCR, the 3' primers were as follows: muCkappa.1 GACATTGATGTCTTTGGGGT (SEQ ID NO:327) and muCkappa.2 TTCACTGCCATCAATCTTCC (SEQ ID NO:328). [0951] The PCR products were separated by agarose gel electrophoresis. The DNA fragments coding for VH and VL were purified using a QIA quick Gel Extraction Kit (obtained from Qiagen), and subcloned into a pCRII-TOPO vector using a TOPO TA cloning kit (obtained from Invitrogen). Each successful clone was sequenced by Sanger sequencing and at least 8 clones were sequenced for each sample.

Antibody purification from hybridoma supernatants

[0952] Hybridoma cells producing each monoclonal antibody were amplified in ClonaCell hybridoma medium A (obtained from StemCell Technologies, Inc.) to a density of 10⁶ cells/mL. Cells were collected by brief centrifugation and washed 3 times with PBS to remove serum. The cells were suspended in Gibico Hybridoma- SFM (obtained from Thermo Fisher Scientific, Inc.) at a density of 10⁶ cells/mL, and cultured with shaking at 200 rpm for 5 days at 37 °C in an atmosphere of 5% CO2. The cell culture medium containing the monoclonal antibody was collected by centrifugation and filtered through a 0.22 μπι filter unit to remove cell debris.

[0953] Antibody was purified by affinity chromatography using Protein A Sepharose from GE Healthcare Life Science. Typically, 50 mL of cell culture supernatant was loaded onto 1 mL of bead slurry for antibody capture. The slurry was washed thoroughly with PBS to remove unbound components, and the bead-bound antibody was eluted with about 5-6X bead volume of elution buffer (i.e., 50 mM sodium citrate, 150 mM sodium chloride, pH 3.0). The elutes were neutralized with 1/5 volume of 1 M arginine pH 5.0 solution to stabilize the antibody. Protein concentration was determined by measuring absorbance at 280 nm (A280), and the purity was tested by gel electrophoresis.

Epitope mapping using peptide ELISA

[0954] Alpha-synuclein peptides were synthesized to represent different fragments of full- length human alpha-synuclein (SEQ ID NO: l), including fragments consisting of amino acids 1-60, 1-90, 1-122, 95-140, and 61-140 of SEQ ID NO: l . Peptides were dissolved to a concentration of 0.5 μg/mL in PBS. 96-well ELISA plates were coated with 50 uL/well of each peptide solution separately and incubated at 4 °C overnight. After 3 washes with 300 μΐ ννεΐΐ of PBST, 150 uL of 5% BSA dissolved in PBS were applied to each well and incubated for 30 minutes at room temperature for blocking, followed by another round of PBST washing. 50 μΐ. of purified antibody solution in PBS, starting at a concentration of 10 μg/mL with a serial dilution factor of four, were applied to each well of the plates. The plates were incubated at room temperature for 1 hour. After three washes with PBST as above, each well then received 50 μΙ_, of 0.2 μ§/ιηΙ. goat anti-mouse IgG conjugated to horseradish peroxidase (obtained from SouthernBiotech, Inc.) dissolved in blocking solution. The plates were incubated for 30 minutes at room temperature. After 3 washes with PBST as above, the antigen-antibody reaction was probed using the TMB Microwell Peroxidase Substrate System (obtained from Kirkegaard & Perry), and color was allowed to develop for 5 minutes. The enzyme reaction was stopped by the addition of 50 μΐ_^ of BioFX TMB-STOP solution (obtained from SurModics, Inc.). OD was measured at 450 nm. Data are shown in Table 9.

Epitope mapping using peptide microarrays

[0955] Full-length alpha-synuclein (SEQ ID NO: 1) was divided into 15-amino acid peptides, offset by 5 amino acids (i.e., overlapping by 10 amino acids). Peptides were synthesized and covalently attached to silica slides in triplicate with a spot size of 0.5 mm (JPT Technologies, Berlin, Germany). Antibodies were diluted to a concentration of 5 μg/mL in 3% bovine serum albumin using Tris-buffered saline (i.e., 10 mM, Tris, pH 7.5, 150 mM NaCl) supplemented with 0.05% Tween 20 (3% BSA-TBST). Diluted antibodies were allowed to bind to the peptides that were printed onto the slides for 2 hours at room temperature as described in the Pepstar user manual (JPT). Following extensive washing (i.e., 5 washes of 5 minutes each with TBST), slides were incubated with secondary antibodies (i.e., donkey anti-mouse IgG, Alexa Fluor^® 647 conjugate, 5 μg/mL in 3% BSA-TBST) for 1 hour at room temperature. Afterward, following extensive washing, (i.e., 5 washes of 5 minutes each with TBST, 5 washes of 5 minutes each with ultrapure water), slides were dried under nitrogen and imaged on an Opera Phenix^™ slide scanner in the 647 nm channel. Images were aligned to a peptide array definition file (Galviewer, obtained from JPT) using ImageJ software with control human IgG serving as landmarks. The sequences of 15-mers that bound to one or more clones are listed in Table 8. Epitopes for clones 3D5, 4F5, 46D2, 28B6, 3E12, 1F4, 1G4, and 16D4 are listed in Table 7.

Biacore^™ kinetics

[0956] Anti-mouse Fc antibody (obtained from GE Healthcare) was immobilized on the surface of a CM5 chip (obtained from GE Healthcare) through amine-coupling to reach about 6,000 to 8,000 response units (RU). The surface was activated by injection of a mixture of 1- ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS), both obtained from GE Healthcare, for 7 minutes. Anti-mouse Fc antibody was diluted in sodium acetate (pH 5.0) at 25 μ§/ιηΙ. and injected for 10 minutes at a flow rate of 5 μΐ,/ιηίηυΐε, followed by injection of ethanolamine (obtained from GE Healthcare) for 7 minutes.

[0957] Purified anti alpha-synuclein antibody (10 μg/mL) was captured to reach 1,500 to 2,000 RU. A range of serially-diluted alpha-synuclein monomer or oligomer (e.g., 3.4 nM to 300 nM) was injected at a flow rate of 30 μΐνπήηυΐε, using either single-cycle kinetics or full- cycle kinetics methods. Sensorgrams were fitted using a 1 : 1 Langmuir model to estimate k₀n and koff. Table 10 provides kinetic parameters for antibodies tested using full-cycle kinetics; these data show that the selected antibodies have affinities ranging from around 4 pM-220 nM. Notably, many of the oligomer specific antibodies have very slow off-rates with koff values less than 4.0xl0^"4 s^"1. Table 1 1 shows the selective binding of antibodies to human alpha-synuclein, cross reactivity to mouse alpha-synuclein, and no binding to structurally similar beta- or gamma-synuclein.

Example 6. ELISA measurement of bound and free alpha-synuclein protein

[0958] This example describes the use of ELISA to measure the amounts of bound and free alpha-synuclein protein.

[0959] The amounts of bound and free alpha-synuclein protein are measured in interstitial fluid (ISF) and cerebrospinal fluid (CSF) samples using a modified commercial Mesoscale Discovery (MSD) protocol. Biotinylated anti-alpha-synuclein capture antibody (rabbit IgG) is used to coat 96-well streptavidin small-spot MSD plates, as per manufacturer' s protocol (e.g., using a human alpha-synuclein U-Plex kit). Plates are incubated with capture antibody for 1 hour at room temperature with shaking (700 rpm). Plates are subsequently washed three times in PBS-Tween using a plate washer. Standardized or unknown samples are added to each well and incubated for 2 hours. Modifications for measuring bound alpha-synuclein and free alpha- synuclein assays are described below.

[0960] For bound alpha-synuclein, alpha-synuclein bound to a therapeutic antibody is measured using a modified version of the protocol above. Briefly, plates are coated and samples are added as described above. Recombinant human alpha-synuclein standards are prepared as per manufacturer's protocol, and 25 uL/well is added to the standard curve wells containing 25 μΕΛνεΙΙ of 1.67 nM therapeutic antibody (separate standard curve for each therapeutic antibody tested; total volume of 50 μΐ ννεΐΐ). 25 μΙ_, test samples are added to 25 μΐ ννεΐΐ of diluent only. Samples and standards are incubated for 2 hours at room temperature with shaking. Plates are washed three times with PBST and then incubated with anti-mouse SULFO-TAG-labeled antibody, using manufacturer's recommended dilutions, at room temperature for 30 minutes with shaking. After secondary antibody incubation, plates are washed three times in PBST and signal is read using lx MSD read buffer. A positive signal indicates the presence of mouse IgG bound to alpha-synuclein in the ISF or CSF.

[0961] Free alpha-synuclein is measured using a modified version of the assay described above. Briefly, plates were coated and samples are added as described above. For measuring free alpha-synuclein, a human IgG version of the therapeutic antibody is generated, which is used as the detection antibody. Since the binding site of the human IgG antibody is the same as the therapeutic antibody (mouse IgG), only free alpha-synuclein will produce a positive signal (i.e., since the binding site of alpha-synuclein bound to the therapeutic antibody is occluded). Anti-human SULFO-TAG-labeled antibody is used as the secondary antibody to measure free alpha-synuclein. 1.67 nM free alpha-synuclein detection antibody (human IgG) is added for 10 minutes only to prevent competition with the therapeutic antibody for alpha- synuclein binding.

Example 7. Anti-alpha-synuclein antibodies comprising modified Fc polypeptides

[0962] This example describes the generation and characterization of anti-alpha-synuclein antibodies that comprise modified Fc polypeptides.

Generation of anti-alpha-synuclein Fabs fused to BBB-penetrating Fc polypeptides

[0963] The Fd (VH+CH1 regions) of antibodies 2F09 and 1F4 (SEQ ID NOs:330 and 331) were cloned into expression vectors comprising a sequence encoding an Fc polypeptide engineered to bind to the transferrin receptor (TfR) (CH3C.35.21.17.2). The Fc polypeptide- encoding sequence also contained a "knob" (T366W) mutation to prevent homodimerization and promote heterodimerization with an Fc polypeptide comprising "hole" (T366S/L368A/Y407V) mutations. The Fd-Fc fusion constructs were designed to express the proteins in SEQ ID NOs:332 and 333. The Fd regions were also cloned into corresponding "hole" vectors comprising a sequence encoding an Fc polypeptide with hole mutations, but lacking the TfR binding mutations, and designed to express the proteins in SEQ ID NOs:334 and 335.

[0964] The corresponding aforementioned knob and hole vectors were co-transfected to ExpiCHO or Expi293 cells along with the corresponding light chain vectors (SEQ ID NOs:617 and 618) in the ratio knob:hole:light chain of 1 : 1 :2. The expressed proteins were purified by Protein A chromatography followed by preparative size-exclusion chromatography (SEC) by methods familiar to those of skill in the art. The full complexes are herein referred to as 2F09/35.21.17.2 and 1F4/35.21.17.2.

Alpha-synuclein/transferrin receptor (TfR) binding

[0965] The binding of bispecific alpha-synuclein/TfR-binding proteins to the respective targets was measured using surface plasmon resonance (SPR) on a Biacore™ T200 instrument. To measure TfR binding, anti-human-Fab was immobilized on a CM5 chip, and the alpha- synuclein/TfR-binding protein was captured. Full-length human TfR or human TfR apical domain at serial dilution (e.g., concentrations of 1-1,000 nM) was flowed over the chip (180 second association time) and then allowed to dissociate. Fitting was performed using a 1 : 1 binding model. To measure alpha-synuclein binding, anti-human-Fc antibody was immobilized on a CM5 chip, and the alpha-synuclein/TfR-binding protein was captured. A range of concentrations of recombinant alpha-synuclein protein were flowed over the chip, and then allowed to associate and dissociate. The resulting sensograms were fit using a 1 : 1 Langmuir model to estimate k₀n and k₀ff. Results are shown in Table 14 below. As is clear from this data, the presence of the TfR-binding Fc region does not impact the ability of the polypeptide to bind alpha-synuclein.

Table 14. Kinetics for Anti-Alpha Synuclein Antibodies and Corresponding Fab's Fused to

Engineered TfR-binding Fc region CH3C.35.21.17.2

Example 8. Modified Fc polypeptides that bind to TfR

[0966] This example describes modifications to Fc polypeptides to confer transferrin receptor (TfR) binding and transport across the blood-brain barrier (BBB).

[0967] Unless otherwise indicated, the positions of amino acid residues in this section are numbered based on EU index numbering for a human IgGl wild-type Fc region. Generation and characterization of Fc polypeptides comprising modifications at positions 384.

386. 387. 388. 389. 390. 413. 416. and 421 (CH3C clones)

[0968] Yeast libraries containing Fc regions having modifications introduced into positions including amino acid positions 384, 386, 387, 388, 389, 390, 413, 416, and 421 were generated as described below. Illustrative clones that bind to TfR are shown in Tables 12 and 13.

[0969] After an additional two rounds of sorting, single clones were sequenced and four unique sequences were identified. These sequences had a conserved Tip at position 388, and all had an aromatic residue (i.e., Tip, Tyr, or His) at position 421. There was a great deal of diversity at other positions.

[0970] The four clones selected from the library were expressed as Fc fusions to Fab fragments in CHO or 293 cells, purified by Protein A and size-exclusion chromatography, and then screened for binding to human TfR in the presence or absence of holo-Tf by ELISA. The clones all bound to human TfR and the binding was not affected by the addition of excess (5 μΜ) holo-Tf. Clones were also tested for binding to 293F cells, which endogenously express human TfR. The clones bound to 293F cells, although the overall binding was substantially weaker than the high-affinity positive control.

[0971] Next, it was tested whether clones could internalize in TfR-expressing cells using clone CH3C.3 as a test clone. Adherent HEK 293 cells were grown in 96-well plates to about 80% confluence, media was removed, and samples were added at 1 μΜ concentrations: clone CH3C.3, anti-TfR benchmark positive control antibody (Ab204), anti-BACEl benchmark negative control antibody (Abl07), and human IgG isotype control (obtained from Jackson Immunore search). The cells were incubated at 37 °C and 8% CO2 concentration for 30 minutes, then washed, permeabilized with 0.1% Triton^™ X-100, and stained with anti-human-IgG-Alexa Fluor^® 488 secondary antibody. After additional washing, the cells were imaged under a high- content fluorescence microscope (i.e., an Opera Phenix^™ system), and the number of puncta per cell was quantified. At 1 μΜ, clone CH3C.3 showed a similar propensity for internalization to the positive anti-TfR control, while the negative controls showed no internalization.

Further engineering of clones

[0972] Additional libraries were generated to improve the affinity of the initial hits against human TfR using a soft randomization approach, wherein DNA oligos were generated to introduce soft mutagenesis based on each of the original four hits. Additional clones were identified that bound TfR and were selected. The selected clones fell into two general sequence groups. Group 1 clones (i.e., clones CH3C.18, CH3C.21, CH3C.25, and CH3C.34) had a semi- conserved Leu at position 384, a Leu or His at position 386, a conserved and a semi-conserved Val at positions 387 and 389, respectively, and a semi-conserved P-T-W motif at positions 413, 416, and 421, respectively. Group 2 clones had a conserved Tyr at position 384, the motif TXWSX at positions 386-390, and the conserved motif S/T-E-F at positions 413, 416, and 421, respectively. Clones CH3C.18 and CH3C.35 were used in additional studies as representative members of each sequence group.

Epitope mapping

[0973] To determine whether the engineered Fc regions bound to the apical domain of TfR, TfR apical domain was expressed on the surface of phage. To properly fold and display the apical domain, one of the loops had to be truncated and the sequence needed to be circularly permuted. Clones CH3C.18 and CH3C.35 were coated on ELISA plates and a phage ELISA protocol was followed. Briefly, after washing and blocking with 1% PBS A, dilutions of phage displaying were added and incubated at room temperature for 1 hour. The plates were subsequently washed and anti-M13-HRP was added, and after additional washing the plates were developed with TMB substrate and quenched with 2N H2SO4. Both clones CH3C.18 and CH3C.35 bound to the apical domain in this assay.

Paratope mapping

[0974] To understand which residues in the Fc domain were most important for TfR binding, a series of mutant clone CH3C.18 and clone CH3C.35 Fc regions was created in which each mutant had a single position in the TfR binding register mutated back to wild-type. The resulting variants were expressed recombinantly as Fab-Fc fusions and tested for binding to human or cynomolgus (cyno) TfR. For clone CH3C.35, positions 388 and 421 were important for binding; reversion of either of these to wild-type completely ablated binding to human TfR.

Binding characterization of maturation clones

[0975] Binding ELISAs were conducted with purified Fab-Fc fusion variants with human or cyno TfR coated on the plate, as described above. The variants from the clone CH3C.18 maturation library, clone CH3C.3.2-1, clone CH3C.3.2-5, and clone CH3C.3.2-19, bound human and cyno TfR with approximately equivalent EC50 values, whereas the parent clones CH3C.18 and CH3C.35 had greater than 10-fold better binding to human versus cyno TfR. [0976] Next, it was tested whether the modified Fc polypeptides internalized in human and monkey cells. Using the protocol described above, internalization in human HEK 293 cells and rhesus LLC-MK2 cells was tested. The variants that similarly bound human and cyno TfR, clones CH3C.3.2-5 and CH3C.3.2-19, had significantly improved internalization in LLC-MK2 cells as compared with clone CH3C.35.

Additional engineering of clones

[0977] Additional engineering to further affinity mature clones CH3C.18 and CH3C.35 involved adding additional mutations to the positions that enhanced binding through direct interactions, second-shell interactions, or structure stabilization. This was achieved via generation and selection from an "NNK walk" or "NNK patch" library. The NNK walk library involved making one-by-one NNK mutations of residues that are near to the paratope. By looking at the structure of Fc bound to FcyRI (PDB ID: 4W40), 44 residues near the original modification positions were identified as candidates for interrogation. Specifically, the following residues were targeted for NNK mutagenesis: K248, R255, Q342, R344, E345, Q347, T359, K360, N361, Q362, S364, K370, E380, E382, S383, G385, Y391, K392, T393, D399, S400, D401, S403, K409, L410, T411, V412, K414, S415, Q418, Q419, G420, V422, F423, S424, S426, Q438, S440, S442, L443, S444, P4458, G446, and K447. The 44 single point NNK libraries were generated using Kunkel mutagenesis, and the products were pooled and introduced to yeast via electroporation, as described above for other yeast libraries.

[0978] The combination of these mini-libraries (each of which had one position mutated, resulting in 20 variants) generated a small library that was selected using yeast surface display for any positions that led to higher affinity binding. Selections were performed as described above, using TfR apical domain proteins. After three rounds of sorting, clones from the enriched yeast library were sequenced, and several "hot-spot" positions were identified where certain point mutations significantly improved the binding to apical domain proteins. For clone CH3C.35, these mutations included E380 (mutated to Tip, Tyr, Leu, or Gin) and S415 (mutated to Glu). The sequences of the clone CH3C.35 single and combination mutants are set forth in SEQ ID NOs:336-340 and 396-402. For clone CH3C.18, these mutations included E380 (mutated to Trp, Tyr, or Leu) and K392 (mutated to Gin, Phe, or His). The sequences of the clone CH3C.18 single mutants are set forth in SEQ ID NOs:390-395. Additional maturation libraries to improve clone CH3C.35 affinity

[0979] An additional library to identify combinations of mutations from the NNK walk library, while adding several additional positions on the periphery of these, was generated as described for previous yeast libraries. In this library, the YxTEWSS (SEQ ID NO:457) and TxxExxxxF (SEQ ID NO:458) motifs were kept constant, and six positions were completely randomized: E380, K392, K414, S415, S424, and S426. Positions E380 and S415 were included because they were "hot spots" in the NNK walk library. Positions K392, S424, and S426 were included because they make up part of the core that may position the binding region, while K414 was selected due to its adjacency to position 415.

[0980] This library was sorted, as previously described, with the cyno TfR apical domain only. The enriched pool was sequenced after five rounds, and the sequences of the modified regions of the identified unique clones are set forth in SEQ ID NOs:341 and 405-421.

[0981] The next libraries were designed to further explore acceptable diversity in the main binding paratope. Each of the original positions (384, 386, 387, 388, 389, 390, 413, 416, and 421) plus the two hot spots (380 and 415) were individually randomized with NNK codons to generate a series of single-position saturation mutagenesis libraries on yeast. In addition, each position was individually reverted to the wild-type residue, and these individual clones were displayed on yeast. It was noted that positions 380, 389, 390, and 415 were the only positions that retained substantial binding to TfR upon reversion to the wild-type residue (some residual but greatly diminished binding was observed for reversion of 413 to wild-type).

[0982] The single-position NNK libraries were sorted for three rounds against the human TfR apical domain to collect the top -5% of binders, and then at least 16 clones were sequenced from each library. The results indicate what amino acids at each position can be tolerated without significantly reducing binding to human TfR, in the context of clone CH3C.35. A summary is below:

Position 380: Trp, Leu, or Glu;

Position 384: Tyr or Phe;

Position 386: Thr only;

Position 387: Glu only;

Position 388: Trp only; Position 389: Ser, Ala, or Val (although the wild type Asn residue seems to retain some binding, it did not appear following library sorting);

Position 390: Ser or Asn;

Position 413 : Thr or Ser;

Position 415: Glu or Ser;

Position 416: Glu only; and

Position 421 : Phe only.

[0983] The above residues, when substituted into clone CH3C.35 as single changes or in combinations, represent paratope diversity that retains binding to TfR apical domain. Clones having mutations at these positions include those shown in Table 13, and the sequences of the CH3 domains of these clones are set forth in SEQ ID NOs:336-372.

Example 9. Additional Fc positions that can be modified to confer TfR binding

[0984] Additional modified Fc polypeptides that bind to transferrin receptor (TfR) were generated having modifications at alternative sites in the Fc region, e.g., at the following positions:

positions 274, 276, 283, 285, 286, 287, 288, and 290 (CH2A2 clones);

positions 266, 267, 268, 269, 270, 271, 295, 297, 298, and 299 (CH2C clones);

positions 268, 269, 270, 271, 272, 292, 293, 294, and 300 (CH2D clones);

positions 272, 274, 276, 322, 324, 326, 329, 330, and 331 (CH2E3 clones); or positions 345, 346, 347, 349, 437, 438, 439, and 440 (CH3B clones).

[0985] Illustrative CH3B clones that bind to TfR are set forth in SEQ ID NOs:430-434. Illustrative CH2A2 clones that bind to TfR are set forth in SEQ ID NOs:435-439. Illustrative CH2C clones that bind to TfR are set forth in SEQ ID NOs:440-444. Illustrative CH2D clones that bind to TfR are set forth in SEQ ID NOs:445-449. Illustrative CH2E3 clones that bind to TfR are set forth in SEQ ID NOs:450-454.

Example 10. Methods

Generation of phage-display libraries

[0986] A DNA template coding for the wild-type human Fc sequence was synthesized and incorporated into a phagemid vector. The phagemid vector contained an ompA or pelB leader sequence, the Fc insert fused to c-Myc and 6xHis epitope tags, and an amber stop codon followed by Ml 3 coat protein pill.

[0987] Primers containing "NNK" tricodons at the desired positions for modifications were generated, where N is any DNA base (i.e., A, C, G, or T) and K is either G or T. Alternatively, primers for "soft" randomization were used, where a mix of bases corresponding to 70% wild- type base and 10% of each of the other three bases was used for each randomization position. Libraries were generated by performing PCR amplification of fragments of the Fc region corresponding to regions of randomization and then assembled using end primers containing Sfil restriction sites, then digested with Sfil and ligated into the phagemid vectors. Alternatively, the primers were used to conduct Kunkel mutagenesis. The ligated products or Kunkel products were transformed into electrocompetent E. coli cells of strain TGI (obtained from Lucigen^®). The E. coli cells were infected with M13K07 helper phage after recovery and grown overnight, after which library phage were precipitated with 5% PEG/NaCl, resuspended in 15%) glycerol in PBS, and frozen until use. Typical library sizes ranged from about 10⁹ to about 10¹¹ transformants. Fc-dimers were displayed on phage via pairing between pill-fused Fc and soluble Fc not attached to pill (the latter being generated due to the amber stop codon before pill).

Generation of yeast-display libraries

[0988] A DNA template coding for the wild-type human Fc sequence was synthesized and incorporated into a yeast display vector. For CH2 and CH3 libraries, the Fc polypeptides were displayed on the Aga2p cell wall protein. Both vectors contained prepro leader peptides with a Kex2 cleavage sequence, and a c-Myc epitope tag fused to the terminus of the Fc.

[0989] Yeast display libraries were assembled using methods similar to those described for the phage libraries, except that amplification of fragments was performed with primers containing homologous ends for the vector. Freshly prepared electrocompetent yeast (i.e., strain EBY100) were electroporated with linearized vector and assembled library inserts. Electroporation methods will be known to one of skill in the art. After recovery in selective SD-CAA media, the yeast were grown to confluence and split twice, then induced for protein expression by transferring to SG-CAA media. Typical library sizes ranged from about 10⁷ to about 10⁹ transformants. Fc-dimers were formed by pairing of adjacently displayed Fc monomers.

General methods for phage selection [0990] Phage methods were adapted from Phage Display: A Laboratory Manual (Barbas, 2001). Additional protocol details can be obtained from this reference.

Plate sorting methods

[0991] Antigen was coated on MaxiSorp^® microtiter plates (typically 1-10 μg/mL) overnight at 4 °C. The phage libraries were added into each well and incubated overnight for binding. Microtiter wells were washed extensively with PBS containing 0.05 % Tween^® 20 (PBST) and bound phage were eluted by incubating the wells with acid (typically 50 mM HC1 with 500 mM KC1, or 100 mM glycine, pH 2.7) for 30 minutes. Eluted phage were neutralized with 1 M Tris (pH 8) and amplified using TGI cells and M13/K07 helper phage and grown overnight at 37 °C in 2YT media containing 50 μg/mL carbenacillin and 50 ug/mL kanamycin. The titers of phage eluted from a target-containing well were compared to titers of phage recovered from a non-target-containing well to assess enrichment. Selection stringency was increased by subsequently decreasing the incubation time during binding and increasing washing time and number of washes.

Bead sorting methods

[0992] Antigen was biotinylated through free amines using HS-PEG4-Biotin (obtained from Pierce^™). For biotinylation reactions, a 3- to 5-fold molar excess of biotin reagent was used in PBS. Reactions were quenched with Tris followed by extensive dialysis in PBS. The biotinylated antigen was immobilized on streptavi din-coated magnetic beads, {i.e., M280- streptavidin beads obtained from Thermo Fisher). The phage display libraries were incubated with the antigen-coated beads at room temperature for 1 hour. The unbound phage were then removed and beads were washed with PBST. The bound phage were eluted by incubating with 50 mM HC1 containing 500 mM KC1 (or 0.1 M glycine, pH 2.7) for 30 minutes, and then neutralized and propagated as described above for plate sorting.

[0993] After three to five rounds of panning, single clones were screened by either expressing Fc on phage or solubly in the E. coli periplasm. Such expression methods will be known to one of skill in the art. Individual phage supernatants or periplasmic extracts were exposed to blocked ELISA plates coated with antigen or a negative control and were subsequently detected using HRP-conjugated goat anti-Fc (obtained from Jackson Immunoresearch) for periplasmic extracts or anti-M13 (GE Healthcare) for phage, and then developed with TMB reagent (obtained from Thermo Fisher). Wells with OD450 values greater than around 5-fold over background were considered positive clones and sequenced, after which some clones were expressed either as a soluble Fc fragment or fused to Fab fragments

General methods for yeast selection

Bead sorting Magnetic-assisted cell sorting (MACS)) methods

[0994] MACS and FACS selections were performed similarly to as described in Ackerman, et al. 2009 Biotechnol. Prog. 25(3), 774. Streptavidin magnetic beads (e.g. , M-280 streptavidin beads from Thermo Fisher) were labeled with biotinylated antigen and incubated with yeast (typically 5-10x library diversity). Unbound yeast were removed, the beads were washed, and bound yeast were grown in selective media and induced for subsequent rounds of selection.

Fluorescence-activated cell sorting (FACS) methods

[0995] Yeast were labeled with anti-c-Myc antibody to monitor expression and biotinylated antigen (concentration varied depending on the sorting round). In some experiments, the antigen was pre-mixed with streptavidin-Alexa Fluor^® 647 in order to enhance the avidity of the interaction. In other experiments, the biotinylated antigen was detected after binding and washing with streptavidin-Alexa Fluor^® 647. Singlet yeast with binding were sorted using a FACS Aria III cell sorter. The sorted yeast were grown in selective media then induced for subsequent selection rounds.

[0996] After an enriched yeast population was achieved, yeast were plated on SD-CAA agar plates and single colonies were grown and induced for expression, then labeled as described above to determine their propensity to bind to the target. Positive single clones were subsequently sequenced for binding antigen, after which some clones were expressed either as a soluble Fc fragment or as fused to Fab fragments.

General methods for screening

Screening by ELISA

[0997] Clones were selected from panning outputs and grown in individual wells of 96-well deep-well plates. The clones were either induced for periplasmic expression using autoinduction media (obtained from EMD Millipore) or infected with helper phage for phage- display of the individual Fc variants on phage. ELISA plates were coated with antigen, typically at 0.5 mg/mL overnight, then blocked with 1% BSA before addition of phage or periplasmic extracts. After a 1-hour incubation and washing off unbound protein, HRP- conjugated secondary antibody was added (i.e., anti-Fc or anti-M13 for soluble Fc or phage- displayed Fc, respectively) and incubated for 30 minutes. The plates were washed again, and then developed with TMB reagent and quenched with 2N sulfuric acid. Absorbance at 450 nm was quantified using a plate reader (BioTek^®) and binding curves were polotted using Prism software where applicable. In some assays, soluble transferrin or other competitor was added during the binding step, typically at significant molar excess.

Screening by flow cytometry

[0998] Fc variant polypeptides (expressed either on phage, in periplasmic extracts, or solubly as fusions to Fab fragments) were added to cells in 96-well V-bottom plates (about 100,000 cells per well in PBS+1%BSA (PBSA)), and incubated at 4 °C for 1 hour. The plates were subsequently spun and the media was removed, and then the cells were washed once with PBSA. The cells were resuspended in PBSA containing secondary antibody (typically goat anti-human-IgG-Alexa Fluor^® 647 (obtained from Thermo Fisher)). After 30 minutes, the plates were spun and the media was removed, the cells were washed 1-2 times with PBSA, and then the plates were read on a flow cytometer (i.e., aFACSCanto™ II flow cytometer). Median fluorescence values were calculated for each condition using FlowJo software and binding curves were plotted with Prism software.

[0999] The amino acid substitutions for each clone described in the Tables (e.g., Table 13) dictate the amino acid substitutions at the register positions of that clone over the amino acids found in the sequence set forth in the Sequence Listing, in case of discrepancy.

[1000] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. The sequences of the sequence accession numbers cited herein are hereby incorporated by reference. Table 1. Unique Hit Sequences

1G03 18 41 65 108 138 185 209 228 237

2A08 18 42 65 106 138 184 209 228 237

2B08 18 43 65 106 138 186 220 228 237

2C07 18 44 65 109 138 187 218 228 237

2D07 18 45 65 110 138 188 212 228 237

2G07 18 46 65 84 138 184 209 228 237

2H07 18 47 65 84 138 189 215 228 237

2D08 18 48 65 89 139 190 218 228 237

2B09 18 49 65 111 140 191 212 226 237

2C09 18 50 77 112 140 192 218 228 237

2D09 18 51 65 113 140 193 209 228 237

2F08 18 52 65 114 140 194 216 227 236

2G08 18 53 65 89 140 195 209 228 237

2H08 18 54 65 87 140 196 209 228 237

2E09 18 55 67 115 141 197 217 228 237

2F09 22 56 79 116 142 198 221 229 238

1E04 23 57 80 117 143 199 218 228 237

2G09 23 57 80 117 143 200 222 228 237

2F10 23 58 65 118 144 201 219 230 237

1G06 30 59 63 119 145 202 220 316 239

3C02 30 60 63 119 145 203 220 228 239

3D02 30 61 63 120 145 204 220 316 239

4F5 hybridoma 269 276 279 285 292 218 228 237

1G4 hybridoma 270 277 281 286 293 301 223 306

16D4 hybridoma 271 65 282 287 294 302 228 237

28B6 hybridoma 272 277 283 288 295 303 305 307

3E12 hybridoma 273 277 280 289 296 304 223 308

3D5 hybridoma 273 277 280 289 297 304 223 308

46D2 hybridoma 274 278 284 290 298 206 224 232

1F4 hybridoma 275 277 283 291 299 304 223 308F4 C^S

variant hybridoma 275 277 283 291 300 304 223 308

Table 2. Biacore Single-Cycle Kinetic Parameters (alpha-synuclein monomer)

Table 3. Biacore Full-Cycle Kinetic Parameters (alpha-synuclein monomer)

full cycle

kon full cycle full cycle

Clone (1/M-s) koff (1/s) Kp (M)

1F06 poor fit poor fit poor fit

2D02 4.70E+04 3.00E-04 6.38E-09

2F09 9.20E+05 1.50E-03 1.63E-09

2F10 1.00E+05 6.70E-04 6.70E-09

3C02 4.10E+04 1.20E-03 2.93E-08

Table 4. Biacore Full-Cycle Kinetic Parameters (alpha-synuclein oligomer)

Table 5. ELISA Binding Data

Table 6. Peptide Array Epitope Mapping for Families 10, 18, 22, 23, and 30

Table 7. Epitope Sequences

Table 8. Peptide Array Epitope Mapping for Hybridoma-Produced Antibodies

46D2 322 EMP SEEGYQD YEPE A

28B6 258 GSIAAATGFVKKDQL

28B6 259 ATGFVKKDQLGKNEE

28B6 323 DNEAYEMPSEEGYQD

28B6 322 EMP SEEGYQD YEPE A

1F4 323 DNEAYEMPSEEGYQD

1F4 322 EMP SEEGYQD YEPE A

1G4 258 GSIAAATGFVKKDQL

1G4 259 ATGFVKKDQLGKNEE

16D4 258 GSIAAATGFVKKDQL

16D4 259 ATGFVKKDQLGKNEE

Table 9. ELISA Epitope Mapping Data

Table 10. Biacore Full-Cycle Kinetic Parameters (alpha-synuclein monomer and oligomer)

Table 11. Binding Data

Table 12. CH3 Domain Modifications

Table 13. Additional CH3 Domain Modifications

Table 15. Informal Sequence Listing

EIQLQQSGPVLVKPGASVKMSCKASGYTFTDYYMNWVKQSHGKSLEWIGFI

VH for clone

HPYSGDARYNQKFKGKATLTVDKSSRTAYMELDSLTSEDSAVYYCTRGDVD

1D02 (family 18)

WGQGTSVTVSP

KVQLQQSGPVLVKPGSSVKMSCKASGYTFTDYYMNWVKQSHGKSLEWIGFI

VH for clone HPYSGDTNYNQNFKGKATLTVDKSSRTAYMELNSLTSEDSAVYYCTRGDVD

2A04 (family 18) WGQGTSVTVSS

EVQLQQSGPVLVKPGASVKMSCQASGFTFTDYYMNWVKQSHGKSLEWIGFI

VH for clone HPYSGDTQYNQKFKGRATLTVDKSSRTAYMELHSLTSEDSAVYYCTRGDVD

2B04 (family 18) WGQGTTLTVSS

EVQLQQSGPVLVKPGASVKMSCKASGYTFTDYYMNWVKQSHGKSLEWIGFI

VH for clone HPYSGDTRYNQKFKGKATLTVDKS SRTAYMELNRLTSED S AVYYCTRGD VD

2C04 (family 18) WGQGTSVTVSS

EVHVKQSGAELVRPGASVKLSCKASGYTFTDYYINWVKQSHGKSLEWIGFIH

VH for clone PYRGDTRYNQKFKGK ATLT VDKS SRT AYMELNSLTSED S AVYYCTRGD VD

2E03 (family 18) WGQGTT VTVS S

QVQLQQPGAELVKPGASVKMSCKASGFTFTDYYMNWVKQSHGKSLEWMG

VH for clone FIHPYTGDISYNQKFKDKATLTVDKSSRTAYMELNSLTSEDSAVYYCTRGDV

2E04 (family 18) DWGQGTTLTVSS

EVQLQQSGPVLVKPGASVKISCKASGYTFTDYYMNWVKQSHGKSLEWIGFI

VH for clone HPYTGDTSYSQKFKDKATLTVDKSSRTAYMELNSLTSEDSAVYYCTRGDVD

2G04 (family 18) WGQGTTLTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITNYYYWNWIRQFPGN LQWMGYI

VH for clone

SYDGNSNYSPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCTSGDDYWG

2H04 (family 18)

QGTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGFSITSGYYWNWIRQFPGNKLEWVGFIS

VH for clone QDGSSNYNPSLKNRFSITRDTSKNQFFLKLSSVTTEDTATYYCVKGDGYWGQ

2B05 (family 18) GTSVTVSS

EVKLQESGPGLVKPSQSLSLSCSVTGYSITSGYYWNWIRQFPGN LEWMGYI

VH for clone SDDGSN YNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVKGDGYWG

2C05 (family 18) QGTTVTVSS

EVQLQESGPGLVKPSQSLSITCSVTGYSITRGYYWNWIRQFPGN LEWMGYI

VH for clone

NYDGSNDYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVKGDVDWG

2D05 (family 18)

QGTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITRGYYWNWIRQFPGN LEWMGYI

VH for clone NYDGSNDYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVKGDVDWG

2E05 (family 18) QGTTVTVSS

EVQLQESGPGLVKPSQSLSLTCSVTGYPITSGYYWNWIRQFPGNKLEWMGYI

VH for clone SDDGSN YNPSLKNRISITRDTSKNQFFLKLNSVTTEDIATYYCVRGDIHWGQ

2G05 (family 18) GTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNTLEWMGFIS

VH for clone

DDGSSNYNPSLKNRISITRDTYKNQFFLKFNSVTTDDTATYYCVRGDILWGQ

2A06 (family 18)

GTLVTVSA

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGFYWNWIRQFPGNTLEWMGFIS

VH for clone DDGSSDYNPSLKNRISITRDTYKNQFFLKFNSVTTDDTATYYCVRGDILWGQ

2H05 (family 18) GTLVTVSA

EVQLQQSGPVLVKPGASVKMSCKASGNTFTDYYMNWVKQSHGQSLEWIGLI

VH for clone HPYNGGTSYAQKFKGKATLTVDKSSRTAYMELNSLTSEDSAVYYCVRGDLA

2C06 (family 18) WGQGTTLTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone NYDGSNDYNPSLKNRISITRDTSQNQFFLKLNSVTSEDTATYYCVRGDVDWG

2G06 (family 18) QGTSVTVSS

EVQLQESGPGLVKPSQSLSITCSVTGYSITRNYYWNWIRQFPGN LEWMAYIS

VH for clone YDGSNSYNPSLKDRISVTRDTSKNQFFLKLSSVTTEDTATYYCVRGDVDWGQ

2H06 (family 18) GTTVTVSS EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGFYWNWIRQFPGNTLEWMGFIS

VH for clone DDGSSNYNPSLKNRISITRDTSKNQFFLKFNSVTTEDTATYYCVRGDVHWGQ

1D03 (family 18) GTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone SDDGSSNYNPSLKNRISITRDTSKNQFFLKFNSVTTEDTATYYCVRGDVHWG

2A07 (family 18) QGTSVTVSS

EVQLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLQWMGYI

VH for clone SDDGSDNYTPSLKNRISITRDTSKNQFFLILNSVTIEDTATYYCVRGDVHWGQ

2B07 (family 18) GTLVTVSA

EVQLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

VH for clone DDGTN YNPSLKNRISITRDTSKNQFFLKLNSVTSEDTATYYCVRGDVVWGT

1E03 (family 18) GTSVTVSS

EVQLQESGPGLVKPSQSPSLTCSVTDYSITNGYYWNWIRQFPGNKLEWMGYI

VH for clone STDGTNNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVRGDWWG

1F03 (family 18) TGTTLTVSS

KVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone TTDGTN YNPSLKNRISITRDTSKNQVFLKLNSVTTEDTATYYCVRGDVVWG

1G03 (family 18) TGTSVTVSS

EVQLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

VH for clone DDGTN YNPSLKNRISITRDTSKNQFFLKLNSVTSEDTATYYCVRGDVVWGT

2A08 (family 18) GTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

VH for clone DDGTN YNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVRGDWWGT

2B08 (family 18) GTTVTVSS

KVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWVRQFPGSKLEWMGYI

VH for clone SDDGSHNYNPSLKNRISITRDTSKNQFFLRLNSVTTEDTATYYCVRGDWWG

2C07 (family 18) TGTTVTVSS

KVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone SDDGSN YNPSLENRISITRDTSKNQFFLKLNSVTTEDTATYYCVRGDWWG

2D07 (family 18) TGTSVTVSS

EVQLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone SDDGSN YNPSLKNRISITRDTSKNQFFLKLNSVTIEDTATYYCVRGDWWG

2G07 (family 18) TGTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone SDDGSN YNPSLKNRISITRDTSKNQFFLKLNSVTIEDTATYYCVRGDWWG

2H07 (family 18) TGTTVTVSS

EVKLQESGLGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGFI

VH for clone

NYDGSNDYNPSLKNRISITRDTSKNQFFLKLNSVTSADTATYYCVRGDVYWG

2D08 (family 18)

QGTSVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQLPGNRLEWMGYI

VH for clone S YD GNTD YNP SLKNRISITRDTSKNQFFLKLNS VTTEDTATYYC VS GD VYWG

2B09 (family 18) QGTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSFYYWNWIRQFPGNKLEWMGYIS

VH for clone YDGSDNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVSGDVYWGQ

2C09 (family 18) GTSVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone SYDGSNDYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVSGDVYWG

2D09 (family 18) QGTTLTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone NYDGSDNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVSGDVYWG

2F08 (family 18) QGTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone NYDGSNDYNPSLKNRISITRDTSQNQFFLKLNSVTTEDTATYYCVSGDVYWG

2G08 (family 18) QGTTVTVSS

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWMGYI

VH for clone NYDGSN YNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATYYCVSGDVYWG

2H08 (family 18) QGTTVTVSS

2G04, and 2E09

(family 18)

CDR-H1 for

GFTFTDYYMN clones 2B04 and

2E04 (family 18)

CDR-H1 for clone

GYSITNYYYWN

2H04 (family 18)

CDR-H1 for clone

GFSITSGYYWN

2B05 (family 18)

CDR-H1 for clones 2D05,

GYSITRGYYWN

2E05, and 1H02 (family 18)

CDR-H1 for clone

GYSIPRGYYWN

2C02 (family 18)

CDR-H1 for

GYSITSGFYWN clones 2H05 and

1D03 (family 18)

CDR-H1 for clone

GNTFTDYYMN

2C06 (family 18)

CDR-H1 for clone

GYSITRNYYWN

2H06 (family 18)

CDR-H1 for clone

DYSITNGYYWN

1F03 (family 18)

CDR-H1 for clone

GYSITSFYYWN

2C09 (family 18)

CDR-H1 for clone

GYTFTDYYIN

2E03 (family 18)

CDR-H1 for clone

GYTFASYAIS

2F09 (family 22)

CDR-H1 for

DYSITSGYYWN clones 1E04 and

2G09 (family 23)

CDR-H2 for

ISSGNNTVYYADTVKG clones 1F06 and

3E02 (family 10)

CDR-H2 for clone

ISSGNDTVYYADTVKD

3F02 (family 10)

CDR-H2 for clones 2D01,

IGDDGSNNYNPSLKN

2E01, 1F02, and 2G02 (family 18)

CDR-H2 for clones 2F01,

ISDDGSNNYNPSLKN 2A03, 2C05,

2G05, 2G07, and 2H07 (family 18)

CDR-H2 for clone

GYISYDGSNNYNPSLKN

2A02 (family 18)

CDR-H2 for

INYDGYNDYTPSLKN clones 1H02 and

2C02 (family 18)

CDR-H2 for

INYDGSNNYNPSLKN clones 2B02 and

2H08 (family 18)

CDR-H2 for clone

ISSDGNSDYNPSLKN

2D02 (family 18)

CDR-H2 for

INYDGSNDYNPSLKN

clones 1A03, 2D05, 2E05,

2G06, 2D08, and

2G08 (family 18)

CDR-H2 for clone

90 IHPYTGDTRYNQRFRG

1D01 (family 18)

CDR-H2 for clone

91 IHPNSGETRNNQKFKG

2C03 (family 18)

CDR-H2 for clone

92 IHPYSGDARYNQKFKG

1D02 (family 18)

CDR-H2 for clone

93 IHPYSGDTNYNQNFKG

2A04 (family 18)

CDR-H2 for clone

94 IHPYSGDTQYNQKFKG

2B04 (family 18)

CDR-H2 for clone

95 IHPYSGDTRYNQKFKG

2C04 (family 18)

CDR-H2 for clone

96 IHPYRGDTRYNQKFKG

2E03 (family 18)

CDR-H2 for clone

97 IHPYTGDISYNQKFKD

2E04 (family 18)

CDR-H2 for clone

98 IHPYTGDTSYSQKFKD

2G04 (family 18)

CDR-H2 for clone

99 ISYDGNSNYSPSLKN

2H04 (family 18)

CDR-H2 for clone

100 ISQDGSSNYNPSLKN

2B05 (family 18)

CDR-H2 for clones 2A06,

101 ISDDGSSNYNPSLKN

lD03, and 2A07 (family 18)

CDR-H2 for clone

102 ISDDGSSDYNPSLKN

2H05 (family 18)

CDR-H2 for clone

103 IHPYNGGTSYAQKFKG

2C06 (family 18)

CDR-H2 for clone

104 ISYDGSNSYNPSLKD

2H06 (family 18)

CDR-H2 for clone

105 ISDDGSDNYTPSLKN

2B07 (family 18)

CDR-H2 for clones 1E03,

106 ISDDGTNNYNPSLKN

2A08, and 2B08 (family 18)

CDR-H2 for clone

107 ISTDGTNNYNPSLKN

1F03 (family 18)

CDR-H2 for clone

108 ITTDGTNNYNPSLKN

1G03 (family 18)

CDR-H2 for clone

109 ISDDGSHNYNPSLKN

2C07 (family 18)

CDR-H2 for clone

110 ISDDGSNNYNPSLEN

2D07 (family 18)

CDR-H2 for clone

111 ISYDGNTDYNPSLKN

2B09 (family 18)

CDR-H2 for clone

112 ISYDGSDNYNPSLKN

2C09 (family 18)

CDR-H2 for clone

113 ISYDGSNDYNPSLKN

2D09 (family 18)

CDR-H2 for clone

114 INYDGSDNYNPSLKN

2F08 (family 18)

CDR-H2 for clone

115 INPHNGGTSYTQKFKG

2E09 (family 18)

CDR-H3 for

136 VRGDVD clones 2G06 and

2H06 (family 18)

CDR-H3 for clones 1D03,

137 VRGDVH

2A07, and 2B07 (family 18)

CDR-H3 for clones 1E03, 1F03, 1G03,

138 VRGDW 2A08, 2B08,

2C07, 2D07, 2G07, and 2H07 (family 18)

CDR-H3 for clone

139 VRGDVY

2D08 (family 18)

CDR-H3 for clones 2B09,

140 VSGDVY 2C09, 2D09,

2F08, 2G08, and 2H08 (family 18)

CDR-H3 for clone

141 VTGDLD

2E09 (family 18)

CDR-H3 for clone

142 ASDYDAY

2F09 (family 22)

CDR-H3 for

143 LRGDNGP clones 1E04 and

2G09 (family 23)

CDR-H3 for clone

144 VRGDYGP

2F10 (family 23)

CDR-H3 for clones 1G06,

145 RSPLRYFDV

3C02, and 3D02 (family 30)

DWMTQTPLSLPVSLGDQASISCRSSQSIVHSNGNTYLEWYLQKPGQSPKLLI

VL for clone 3F02

146 YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPPTFGGG

(family 10)

TKLELK

DVQMTQTPLSLPVSLGDQASISCRSSQSIVHSNGNTYLEWYLQKPGQSPKLLI

VL for clone 1F06

147 YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPPTFGAG

(family 10)

TKLELK

DWMTQTPLSLPVSLGDQASISCRSSQSIVHSNGNTYLEWYLQKPGQSPKLLI

VL for clone 3E02

148 YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPPTFGAG

(family 10)

TKLELK

DVLMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIH

VL for clone

149 LVSKLDSGVPDRFNGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

2D01 (family 18) TRLEIK

DIVLTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSK

VL for clone 2E01

150 LASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEI

(family 18) K

DIVLTQTPLTLSVTIGQPASISCKSSRSLLDSDGETYLNWLLQRPGQSPKRLIYL

VL for clone 2F01

151 VSKLDSGVPDRFTGSGSGTEFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGT

(family 18)

KLEIK

DIVLNQTPLTLSVAIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone 1F02

152 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

(family 18) TKLEIK

DILMTQTPLTLSVTIGQPASISCKSSRSLLDHDGETYLNWLLQRPGQSPKRLIY

VL for clone

153 LVSKLDSGVPDRFTGIGSGTEFTLKNSRVEAEDLGVYYCWQGTHFPQTFGGG

2A02 (family 18) TRLEIK DIVMNQTPLTLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIN

VL for clone

154 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

1H02 (family 18) TKLELK

DVLMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone

155 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2B02 (family 18) TKLELK

DIVLTQTPLTLSVTIGQPASISCKSSRSLLDSDGETYLNWLFQRPGQSPKRLIYL

VL for clone

156 VSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGT

2C02 (family 18) KLEIK

DVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLI VL for clones

157 YLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG 2D02 and 2E03

GTKLELK (family 18)

DILMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone

158 LVSKLDSGVPDRFTGSGSGTDFKLKISRVEAEDLGVYYCWQGTHFPQTFGGG

1A03 (family 18)

TRLEIK

DIVLTQTPLTLSVTIGQPASISCKSSQSLLDIDGETYLNWLLQRPGQSPKRLIYL

VL for clone

159 VSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGT

2A03 (family 18) KLELK

DIVLNQTPLTLSVTIGQPASISCKSSQSLLDSDGKTYLNWLFQRPGQSPKRLIY

VL for clone

160 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2G02 (family 18) TKLEIK

DIVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone

161 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGIYYCWQGTHFPQTFGGGT

1D01 (family 18)

KLELK

DIVLNQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone

162 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2C03 (family 18) TRLEIK

DIVMTQTPLTLSVTIGQPASISCKTSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone

163 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGEG

1D02 (family 18) TKLELK

DIVMTQAPLTLSITIGQPASISCKSSQSLLDRDGETYLNWLLQRPGQSPKRLIY

VL for clone

164 LVSKLDSGVPDRFTGSGSGTDFTLKISSVETEDLGVYYCWQGTHFPQTFGGG

2A04 (family 18) TKLEIKRTVAAPSV

DWMTQFPPTLSVTIGQPAFISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLI

VL for clone

165 HLVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGG

2B04 (family 18)

GTKLELK

DIVLNQTPLTLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIN

VL for clone

166 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

2C04 (family 18) TRLEIK

DIVLNQTPLTLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIY

VL for clone 2E04

167 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

(family 18) TKLELK

DVLMTQTPLTLSVTIGQPAFISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLI

VL for clone

168 HLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG

2G04 (family 18)

GTKLEIK

DVVMTQTPLTLSVTIGQPAFISCKSNQSLLDYDGETYLNWLFQRPGQSPKRLI

VL for clone

169 HL VSKLD SGVPDRFNGGGS GTDFTLKI SRVEAEDLGL YYCWQGTHFPQTFGG

2H04 (family 18) GTKLEIK

DIVLNQTPLTLSVTIGQPASISCKSSQSLLDSDGKTYLNWLLQRPGQSPKRLIY

VL for clone

170 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2B05 (family 18) TKLEIK

DIVMTQAPLTLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIN

VL for clone

171 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

2C05 (family 18) TKLEIK

DIVMTQSHKFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKALIYSA

VL for clone

172 SYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCQQYNSYPRTFGGGTKL

2D05 (family 18) ELK DIVLNQTPLTLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIY

VL for clone 2E05

173 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

(family 18) TKLEIK

DVVMTQTPLTLSVTIGQPAFISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLI

VL for clone

174 HLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG

2G05 (family 18) GTRLEIK

DIVMTQIPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIYL

VL for clone

175 VSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGT

2A06 (family 18) KLELK

DIVLTQAPLTLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIY

VL for clone

176 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGSHFPQTFGGG

2H05 (family 18) TRLEIK

DIVMNQTPLSLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIH

VL for clone

177 LVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2C06 (family 18) TKLEIK

DIVMTQSPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLLRPGQSPKRLIY

VL for clone

178 LVSKLDSGVPDRFTGSGSGTDFTLKVSRVEAKDLGVYYCWQGTHFPQTFGG

2G06 (family 18) GTKLEIK

DVVMTQTPLTLSVTIGQPASISCKSSQSLLDRDGETYLNWLLQRPGQSPKRLI

VL for clone

179 YLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG

2H06 (family 18)

GTKLEIK

DVLMTQTPLSLSVTIGQPASISCKSSQSLLDSDGKTYLNWLLQRPGQSPKRLIY

VL for clone

180 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

1D03 (family 18)

TRLEIK

DVLMTQTPLTLSVTIGQPASISCKSSQSLLDNDGETYLNWLLQRPGQSPKRLM

VL for clone

181 YLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG

2A07 (family 18) GTKLELK

DIVLNQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSK

VL for clone

182 LASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLEI

2B07 (family 18) K

DIVMNQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLIY

VL for clone 1E03

183 LVSKLDSGVPDRFSGSGSGTDFTLKISRVEADDLGLYYCWQGTHFPQTFGGG

(family 18) TKLEIK

DIVMTQTPLTLSVTIGQPAFISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLIH VL for clones

184 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG 1F03, 2A08, and TKLEIK 2G07 (family 18)

DVLMTQTPLSLSVTIGQPASISCKSNQSLLDSDGETYLNWLFQRPGQSPKRLrY

VL for clone

185 LVSKLDSGVPDRFTGGGSGTDFTLKISRVEAEDLGLYYCWQGTHFPQTFGGG

1G03 (family 18) TKLEIK

DILMTQTPLTLSVTIGQPASISCKSSQSLLDSDGKTYLNWLLQRPGQSPKRLIH

VL for clone

186 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGFYYCWQGTHFPQTFGGG

2B08 (family 18) TKLEIK

DVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLI

VL for clone

187 YLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG

2C07 (family 18)

GTRLEIK

DIVMTQAPLSLSVTIGQPASISCKSSQSLLDRDGETYLNWLLQRPGQSPKRLrY

VL for clone

188 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2D07 (family 18) TKLEIK

DIVMTQTPLTLSVTIGQPASISCKSSQSLLDHDGETYLNWLLQRPGQSPKRLIY

VL for clone

189 LVSKLDSGVPDRFTGNGSGTDFTLKISRVEADDLGVYYCWQGTHFPQTFGGG

2H07 (family 18) TKLEIK

DVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLFQRPGQSPKRLIN

VL for clone

190 LVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGG

2D08 (family 18) TKLELK

DVLMTQTPLTLSITIGQPASISCKSSQSLLDRDGETYLNWLLQRPGQSPKRLIY

VL for clone

191 LVSQLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGEG

2B09 (family 18) TKLEIK

CDR-L2 for clone

225 SASYRYS

2D05 (family 18)

CDR-L2 for clone

226 LVSQLDS

2B09 (family 18)

CDR-L2 for clone

227 MSTRASG

2F08 (family 18)

CDR-L2 for clones 2D01,

2F01, 1F02,

2A02, 1H02,

2B02, 2C02,

1A03, 2A03,

2G02, 1D01,

2C03, 1D02,

2A04, 2B04,

2C04, 2E03,

2E04, 2G04,

2H04, 2B05,

2C05, 2E05,

2G05, 2A06,

2H05, 2C06,

2G06, 2H06,

228 LVSKLDS

1D03, 2A07, 1E03, 1F03, 1G03, 2A08, 2B08, 2C07, 2D07, 2G07, 2H07, 2D08, 2C09, 2D09, 2G08, 2H08, 2D02, and 2E09 (family 18), clones 1E04 and 2G09 (family 23), clone 3C02 (family 30), and clones 16D4 and 4F5

CDR-L2 for clone

229 RISNRFS

2F09 (family 22)

CDR-L2 for clone

230 LVSKQDS

2F10 (family 23)

CDR-L3 for clones 1F06,

231 FQGSHVRPT

3E02, and 3F02 (family 10)

CDR-L3 for clone

232 QQWSSNPPT 2E01 (family 18) and clone 46D2

CDR-L3 for clone

233 QQWSSNPLT

2B07 (family 18)

CDR-L3 for clone

234 QQY SYPRT

2D05 (family 18)

CDR-L3 for clone

235 WQGSHFPQT

2H05 (family 18)

CDR-L3 for clone

236 MQHLEYPFT

2F08 (family 18) CDR-L3 for clones 2D01,

2F01, 1F02,

2A02, 1H02,

2B02, 2C02,

1A03, 2A03,

2G02, 1D01,

2C03, 1D02,

2A04, 2B04,

2C04, 2E03,

2E04, 2G04,

2H04, 2B05,

2C05, 2E05,

2G05, 2A06,

2C06, 2G06,

237 WQGTHFPQT 2H06, 1D03,

2A07, 1E03, 1F03, 1G03, 2A08, 2B08, 2C07, 2D07, 2G07, 2H07, 2D08, 2B09, 2C09, 2D09, 2G08, 2H08, 2D02 and 2E09 (family 18), clones 1E04, 2G09, and 2F10 (family 23), and clones 4F5 and 16D4

CDR-L3 for clone

238 SQSTHVPLT

2F09 (family 22)

CDR-L3 for clones 1G06,

239 WQGSHFPLT

3C02, and 3D02 (family 30)

Family 10 CDR-

240 GFxFSDYGMH Hl consensus sequence

Family 18 CDR-

241 xYxIxxxxY Hl consensus sequence

Family 18 CDR-

242 GxTFTDYYxN Hl consensus sequence

Family 23 CDR-

243 xYSITSGYY Hl consensus sequence

Family 10 CDR-

244 ISSGNxTVYYADTVKx H2 consensus sequence

Family 18 CDR-

245 IxxD GxxxYxP SLxx H2 consensus sequence

Family 18 CDR-

246 IxPxxGxxxxxQxFxx H2 consensus sequence Family 23 CDR-

247 ISDDGRNxYNPSLKN H2 consensus sequence

Family 30 CDR-

248 ISRGS SAIYYAD AVxG H2 consensus sequence

Family 18 CDR-

249 xxGDxx H3 consensus sequence

Family 23 CDR-

250 xRGDxGP H3 consensus sequence

Family 18 CDR-

251 xxxxSLLxxDGxTYLN Ll consensus sequence

Family 23 CDR-

252 KSxQSLxDxDGxTYLN Ll consensus sequence

Family 18 CDR-

253 xSxxxS L2 consensus sequence

Family 23 CDR-

254 VSKxDS L2 consensus sequence

Family 18 CDR-

255 xQxxxxPxT L3 consensus sequence

256 GAPQEGILEDMPVDP 1F06 epitope

257 GILEDMPVDPDNEAY 1F06 epitope

2D02, 2F10, 3C02, 4F5, 28B6,

258 GSIAAATGFVKKDQL

1G4, and 16D4 epitope

2D02, 2F10, 4F5,

259 ATGFVKKDQLGKNEE 28B6, 1G4, and

16D4 epitope

260 VGGAWTGVTAVAQK 2F09 epitope

2D02 and 2F09

261 VTGVTAVAQKTVEGA

epitope

262 TVEGAGSIAAATGFV 2F10 epitope

263 GVLYVGSKTKEGWH 3C02 epitope

264 KKDQLGKNEEGAPQE 3C02 epitope

2F10 and 3C02

265 GKNEEGAPQEGILED

epitope

EVKLVESGGGLVKPGGSLKLSCAASGFIFSDYGMHWVRQAPEKGLEWIAFIS VH for clone

266 S GNNTVYYADTVKGPJTISPJ) S AKKTLFLQMTSLRSEDTAMYYCVRH SNFM 1F06 C99M

DYWGQGTSVTVSS variant (family 10)

CDR-H3 for clone

267 VRHSNFMDY 1F06 C99M

variant (family 10)

Family 10 CDR-

268 VRHSNFxDY H3 consensus sequence

DVRLQESGPGLVKPSQSLSLTCSVTGYSITSGHYWNWIRQVPGDKLEWMAFI

SDDGIN YNPSLKNRISISRDTSKNQFFLKLNSVTTEDTATYYCVRGDNLWG

269 VH for clone 4F5

QGTTLTVSS EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

270 RSKSN YATYYADSVKDPJTISPJ)DSESMLFLQMNNLKTEDTAVYYCVRQG VH for clone 1G4 YGINYYAMDYWGQGTSVTVSS

DVQLQESGPGLVKPSQSLALTCSVSGYSITSGYYWNWIRQFPGNKLEWMGY

VH for clone

271 LTYDGNKNYNPSLKNRISITPJ)TSKNQFFLKLNSVTSEDTATYYCVYNTYYW

16D4

GRGTTLTVSS

EVQLVESGGGLVQPEGSLKLSCTASGFSFNTYAMNWVRQAPGKGLEWVARI

VH for clone

272 RmSN YATYYADSVKDPJTISPJ^DSENMLYLQMKKLKTEDTAMYYCVRQ

28B6

GITYYAMDYWGQGTSVTVSS

EVQLVESGGGLVQPKGSLKLSCSASGFSFNTYAMNWVRQAPGKGLEWIVRI

VH for clones

273 RSKSNNYAIYYADSVKDRFTISRDDSESMLYLQMNHLKSEDTAIYYCVRQGI

3E12 and 3D5 TYYSMD YWGQGTS VTVS S

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

VH for clone

274 RSKISNYATYYADSVKDRFTISRDDSQSMLYLQMN LKTEDTAMYYCVTGS

46D2

SYGWFAYWGQGTLVTVSA

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

275 RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI VH for clone 1F4 DFY AMD YWGQGTS VTVS S

CDR-H1 for clone

276 GYSITSGHYWN

4F5

CDR-H1 for clones 1G4, 28B6,

277 GFSFNTYAMN

3E12, 3D5, and 1F4

CDR-H1 for clone

278 GFTFNTYAMH

46D2

CDR-H2 for clone

279 FISDDGIN YNPSLKN

4F5

CDR-H2 for

280 RIRSKSN YAIYYAD SVKD clones 3E12 and

3D5

CDR-H2 for clone

281 RIRSKSN YATYYAD S VKD

1G4

CDR-H2 for clone

282 YLTYDGNKNYNPSLKN

16D4

CDR-H2 for

283 RIRTKSNNYATYYAD SVKD clones 28B6 and

1F4

CDR-H2 for clone

284 RIRSKISNYATYYADSVKD

46D2

CDR-H3 for clone

285 VRGDNL

4F5

CDR-H3 for clone

286 VRQGYGINYYAMDY

1G4

CDR-H3 for clone

287 VYNTYY

16D4

CDR-H3 for clone

288 VRQGITYYAMDY

28B6

CDR-H3 for clone

289 VRQGITYYSMDY

3E12 and 3D5

CDR-H3 for clone

290 VTGSSYGWFAY

46D2

CDR-H3 for clone

291 VRQGIDFYAMDY

1F4

DVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPRRLIY

292 LVSKLDSGVPDRFTGSGSGTDFTLRISRVEAEDLGVYYCWQGTHFPQTFGGG VL for clone 4F5

TKLEIK DVLMTQTPLSLPVSLGDQASISCRSSQSIVHSYGSTYLEWYLQKPGQSPKLLIY

293 KVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPLTFGAGT VL for clone 1G4 KLELK

DVVMTQTPLTLSVIIGQPASISCKSSQSLLDVDGETYLNWVLQRPGQSPKRLIS

VL for clone

294 LVSKLDSGVPDRFTGSGSGTDFTLKIIRVEAEDLGVYYCWQGTHFPQTFGGG

16D4

TQLEIK

DVLMTQTPLSLTVSVGDQASISCRSGQSILHSNGNTYLEWYLQKPGQSPNLLI YKVSTRFSGVPDRFSGSGSGTEFTLKISRVEAEDLGVYYCFQGSRGPLTFGEG VL for clone

295

TKLELK 28B6

DVLMTQSPLSLPVSLGDQASISCRSSRDIVHSNGNTYLEWYLQKPGQSPKLLI

296 YKVSNRFSGVPDRFSGSGSETDFTLKISRVEAEDLGVYFCFQGTLIPLTFGAGT VL for clone 3E12

KLELK

DVLMTQSPLSLPVSLGDQASISCRSSRDIVHSNGNTYLEWYLQKPGQSPKLLI

297 YKVSNRFSG DRFSGSGSGTDFTLKISRVEAEDLGVYFCFQGTLIPLTFGAGT VL for clone 3D5

KLELK

QIVLTRSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSK

VL for clone

298 LASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGAGTKLEL

46D2

K

DVLMTQSPLSLSVSLGDQASISCRSSRDIVHSNGNTYLEWYLQKPGQSPKLLI

VL for clone 1F4

299 YKVSNRFSGVPDRFCGSGSGTDFTLKISRVEAEDLGVYYCFQGTLIPLTFGAG

TKLELK

DVLMTQSPLSLSVSLGDQASISCRSSRDIVHSNGNTYLEWYLQKPGQSPKLLI

VL for clone 1F4

300 YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGTLIPLTFGAG

C- S variant TKLELK

CDR-L1 for clone

301 RSSQSIVHSYGSTYLE

1G4

CDR-L1 for clone

302 KS SQSLLD VDGETYLN

16D4

CDR-L1 for clone

303 RSGQSILHSNGNTYLE

28B6

CDR-L1 for

304 RSSRDIVHSNGNTYLE clones 3E12, 1F4, and 3D5

CDR-L2 for clone

305 KVSTRFS

28B6

CDR-L3 for clone

306 FQGSHVPLT

1G4

CDR-L3 for clone

307 FQGSRGPLT

28B6

CDR-L3 for

308 FQGTLIPLT clones 3E12, 3D5, and 1F4

CDR-H1

309 GYSITSGxYWN consensus

sequence

CDR-H1

310 GFxFNTYAMx consensus

sequence

CDR-H2

311 RIRxKxxNYATYYAD S VDK consensus

sequence

CDR-H2

312 xxxxDGxxNYNPSLKN consensus

sequence

CDR-L1

313 xSxxxxxxxxGxTYLx consensus

sequence CDR-L2

314 xxSxxxS consensus

sequence

CDR-L3

315 xQxxxxPxT consensus

sequence

CDR-L2 for

316 QVSKLDS clones 1G06 and

3D02 (family 30)

Family 18 CDR-

317 xYxITxxxYWN Hl consensus sequence

Family 18 CDR-

318 GxTFTDYYxN Hl consensus sequence

Family 23 CDR-

319 xYSITSGYYWN Hl consensus sequence

Family 23 CDR-

320 LVSKxDS L2 consensus sequence

EQVTOVGGAVWGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQE 3E12 and 3D5

321

GILEDMPVDPDNEAYEMPSEEGYQDYEPEA epitope

46D2, 28B6, and

322 EMPSEEGYQDYEPEA

1F4 epitope

28B6 and 1F4

323 DNEAYEMPSEEGYQD

epitope mulgGI VH PCR

324 GGACAGGGATCCAGAGTTCC

primer muIgG2 VH PCR

325 AGCTGGGAAGGTGTGCACAC

primer muIgG3 VH PCR

326 CAGGGGCCAGTGGATAGAC

primer muCkappa.1 VL

327 GACATTGATGTCTTTGGGGT

PCR primer muCkappa.2 VL

328 TTCACTGCCATCAATCTTCC

PCR primer

Human IgGl

329 EPKSCDKTHTCPPCP

hinge sequence

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

330 GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN 2F09-Fd

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKS CDKTH

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

331 DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH KP SNTK VDKKVEPKS CDKTH

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD 2F09-Fd fused to

332 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD Fc with TfR

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN binding and knob

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC

LVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ

GFVFSCSVMHEALHNHYTQKSLSLSPGK EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH 1F4-Fd fused to

333 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE Fc with TfR

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV binding and knob

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLWCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

334 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

Fc with hole

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVEWESNGQPEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

1F4-Fd fused to

335 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

Fc with hole

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTV

DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

336 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.20 EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF S CS VMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

337 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21

TEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

Clone

338 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.22 TEWSNYKTTPPVLDSDGSFFLYSKLTVTKSEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

339 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.23

EWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

Clone

340 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.24 TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

Clone

341 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.17 EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF VFS CS VMHEALHNHYT QKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

342 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.20.1 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESFGT EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF S CS VMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

343 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.20.2

EWASYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

344 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.20.3

EWVSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

345 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.20.4

EWSSYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

Clone

346 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESFGT

CH3C.35.20.5 EWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

Clone

347 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESFGT

CH3C.35.20.6 EWVSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

Clone

348 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESFGT

CH3C.35.21.a. l EWS S YKTTPP VLD SD GSFFL YSKLT VTXEEWQQGF VFS CS VMHEALHNHYT QKSLSLSPGK

Clone

349 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.a.2 TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

Clone

350 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.a.3 TEWVSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

351 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.a.4

TEWSSYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

352 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESFGT

CH3C.35.21.a.5

EWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

353 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESFGT

CH3C.35.21.a.6

EWVSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

354 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.1 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESFGT EWSNYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

355 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.23.2 EWANYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

Clone

356 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.23.3 EWVNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

357 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.23.4

EWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

Clone

358 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESFGT

CH3C.35.23.5 EWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

Clone

359 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESFGT

CH3C.35.23.6 EWVNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

360 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESFGT

CH3C.35.24.1

EWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

Clone

361 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.24.2 TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

Clone

362 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.24.3 TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

Clone

363 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.24.4 TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

364 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESFGT

CH3C.35.24.5

EWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

365 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESFGT

CH3C.35.24.6

EWVNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

366 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21.17.1 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESFGT EWS S YKTTPP VLD SD GSFFL YSKLT VTXEEWQQGF S CS VMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

367 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.17.2

EWASYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

368 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.17.3

EWVSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

369 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.17.4

EWSSYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

Clone

370 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESFGT

CH3C.35.21.17.5 EWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

Clone

371 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESFGT

CH3C.35.21.17.6 EWVSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clones

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.N390

372 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

and

EWSNYKTTPPVLDSDGSFFLYSKLTVTKSEWQQGFVFSCSVMHEALHNHYT

CH3C.35.N163

QKSLSLSPGK

373 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGL Clone CH3C.1 VWVGYKTTPPVLDSDGSFFLYSKLTVAKSTWQQGWVFSCSVMHEALHNHY TQKSLSLSPGK

374 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT Clone CH3C.2 VWSHYKTTPPVLDSDGSFFLYSKLTVSKSEWQQGYVFSCSVMHEALHNHYT QKSLSLSPGK

375 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT Clone CH3C.3 EWSQYKTTPPVLDSDGSFFLYSKLTVEKSDWQQGHVFSCSVMHEALHNHYT QKSLSLSPGK

376 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESVGT Clone CH3C.4 PWALYKTTPPVLDSDGSFFLYSKLTVLKSEWQQGWVFSCSVMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

377 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT Clone CH3C.17

VWSKYKTTPPVLDSDGSFFLYSKLTVSKSEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

378 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Clone CH3C.18 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH VWAVYKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

379 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGL Clone CH3C.21 VWVGYKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

380 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESMG Clone CH3C.25

HVWVGYKTTPPVLDSDGSFFLYSKLTVDKSTWQQGWVFSCSVMHEALHNH

YTQKSLSLSPGK

381 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGL Clone CH3C.34 VWVFSKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

382 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT Clone CH3C.35

EWSSYKTTPPVLDSDGSFFLYSKLTVTKSEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

383 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT Clone CH3C.44

EWSNYKTTPPVLDSDGSFFLYSKLTVSKSEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

384 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH Clone CH3C.51

VWVGYKTTPPVLDSDGSFFLYSKLTVSKSEWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

385 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH Clone CH3C.3.1-3

VWVATKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

386 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGP Clone CH3C.3.1-9

VWVHTKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

387 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH Clone CH3C.3.2-5

VWVDQKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone CH3C.3.2-

388 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH

19

VWVNQKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

389 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH Clone CH3C.3.2-1 VW VNFKTTPP VLD SD GSFFLYSKLTVPKSTWQQGWVF SCS VMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone CH3C.18

390 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

variant

KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESLG HVWAVYKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNH

YTQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone CH3C.18

391 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESLGH

variant

VWAVYKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone CH3C.18

392 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVYWESLGH

variant

VWAVYKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone CH3C.18

393 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH

variant

VWAVYQTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

Clone CH3C.18

394 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH

variant VWAVYFTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone CH3C.18

395 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH

variant

VWAVYHTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

Clone

396 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESLG

CH3C.35.13 HVWAVYKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNH YTQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

397 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH

CH3C.35.14

VWAVYQTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

Clone

398 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESLG

CH3C.35.15 HVWAVYQTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNH YTQKSLSLSPGK

Clone

399 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESLG

CH3C.35.16 HVWVNQKTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNH YTQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

400 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESLGH

CH3C.35.17

VWVNQQTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNHY

TQKSLSLSPGK

Clone

401 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESLG

CH3C.35.18 HVWVNQQTTPPVLDSDGSFFLYSKLTVPKSTWQQGWVFSCSVMHEALHNH YTQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

402 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.19 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG TEWSSYKTTPPVLDSDGSFFLYSKLTVT SEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

403 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT

CH3C.35.K165Q

EWSSYQTTPPVLDSDGSFFLYSKLTVT SEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Clone

404 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESYGT CH3C.35.N163.

EWSNYQTTPPVLDSDGSFFLYSKLTVT SEWQQGFVFSCSVMHEALHNHYT K165Q

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

405 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.1

EWSSYKTTPPVLDSDGSFFLYSKLTVTKSEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

406 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.2

EWSSYRTTPPVLDSDGSFFLYSKLTVTKSEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

407 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.3

EWSSYRTTPPVLDSDGSFFLYSKLTVTREEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

408 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.4

EWSSYRTTPPVLDSDGSFFLYSKLTVTGEEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

Clone

409 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.5 EWSSYRTTPPVLDSDGSFFLYSKLTVTREEWQQGFVFSCWVMHEALHNHYT QKSLSLSPGK

Clone

410 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.6 EWSSYRTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCWVMHEALHNHYT QKSLSLSPGK

Clone

411 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.7 EWS S YRTTPP VLD SD GSFFL YSKLT VTTIEEWQQGF TCWVMHEALHNHYT QKSLSLSPGK

Clone

412 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.8 EWS S YRTTPP VLD SD GSFFL YSKLT VTTIEEWQQGF VFTCG VMHEALHNHYT QKSLSLSPGK

Clone

413 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.9 EWS S YRTTPP VLD SD GSFFL YSKLT VTREEWQQGFVFECW VMHEALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

414 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21.10 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT EWS S YRTTPP VLD SD GSFFLYSKLTVTREEWQQGF KCWVMHE ALHNHYT

QKSLSLSPGK

Clone

415 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21. il EWS S YRTTPP VLD SD GSFFL YSKLT VTTEEWQQGF VFKCWVMHE ALHNHYT QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEWCVYVDVSHEDPEWENWYVDGV EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

416 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.12 TEWSSYRTTPPVLDSDGSFFLYSKLTVTREEWQQGFVFSCSVMHE ALHNHYT QKSLSLSPGK

Clone

417 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.13 TEWSSYRTTPPVLDSDGSFFLYSKLTVTGEEWQQGFVFSCSVMHE ALHNHYT QKSLSLSPGK

Clone

418 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.14 TEWSSYRTTPPVLDSDGSFFLYSKLTVTREEWQQGFVFTCWVMHEALHNHY TQKSLSLSPGK

Clone

419 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.15 TEWSSYRTTPPVLDSDGSFFLYSKLTVTGEEWQQGFVFTCWVMHEALHNHY TQKSLSLSPGK

Clone

420 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVWWESYG

CH3C.35.21.16 TEWSSYRTTPPVLDSDGSFFLYSKLTVTREEWQQGFVFTCGVMHEALHNHY TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEWCVYVDVSHEDPEWENWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Clone

421 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVLWESYGT

CH3C.35.21.18

EWSSYRTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHYTQ

KSLSLSPGK

APELLGGPS LFPPK^KBTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Clone

422 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYG CH3C.35.21 with TEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHE ALHNHYT knob mutation QKSLSLSPGK

APEAAGGPS LFPPKPKDTLMISRTPEVTCVYVOVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21 with

423 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYG

knob and LALA TEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHE ALHNHYT

mutations QKSLSLSPGK

APELLGGPS LFPPK^KBTLYITPJiPEWCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21 with

424 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYG

knob and YTE TEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHE ALHNHYT

mutations QKSLSLSPGK

APEAAGGPS LFPPK^KBTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21 with

425 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYG

knob, LALA, and TEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHE ALHNHYT YTE mutations QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEWCVYVDVSHEDPEWENWYVDGV Clone

426 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21 with KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVWWESYG hole mutations TEWSSYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21 with

427 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVWWESYG

hole and LALA

TEWSSYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT

mutations

QKSLSLSPGK

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21 with

428 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVWWESYG

hole and YTE

TEWSSYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

mutations

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21 with

429 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVWWESYG

hole, LALA, and

TEWSSYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT YTE mutations

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

430 KTISKAKGQPRFDYVTTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH3B.1

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYG

FHDLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

431 KTISKAKGQPPJDMVTTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG Clone CH3B.2

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

GFHDLSLSPGK

432 KTISKAKGQPPJEYWTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH3B.3 PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYG FHDLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

433 KTISKAKGQPPJEMWTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH3B.4

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYG

FHDLSLSPGK

434 KTISKAKGQPRFELVTTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH3B.5 PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYG FHDLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVEFIWYVDGVD VRYEWQLPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK

435 TISKAKGQPP^PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP Clone CH2A2.1 EN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVGFVWYVDGV

PVSWEWYWPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI

436 EKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG Clone CH2A2.2

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVQFDWYVDGV

MVRREWHRPPJiEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI

437 EKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG Clone CH2A2.3

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVSFEWYVDGVP

438 VRWEWQWPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Clone CH2A2.4 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVAFTWYVDGV

PWWEWQNPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI

439 EKTISKAKGQPP^PQ TLPPSRDELT NQVSLTCLVKGFYPSDIAVEWESNG Clone CH2A2.5

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDPQTPPWEVKFNWYVDGV

EVHNAKTKPP^EEYYTYYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

440 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2C.1

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDPPSPPWEVKFNWYVDGV

EVHNAKTKPREEEYYSNYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

441 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2C.2

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDPQTPPWEVKFNWYVDGV

EVHNAKTKPREEEYYSNYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

442 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2C.3

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDFRGPPWEVKFNWYVDGV

EVHNAKTKPREEEYYHDYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

443 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2C.4

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDPQTVPWEVKFNWYVDGV

EVHNAKTKPREEEYYSNYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

444 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2C.5

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSVPPRMVKFNWYVDGV

EVHNAKTKSLTSQHNSTVRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

445 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2D.1

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEWCVVVDVS PWMVKFNWYVDG

VEVHNAKTKSLTSQHNSTVRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI

446 EKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG Clone CH2D.2

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEWCVVVDVSDMWEYVKFNWYVDG

VEVHNAKTKPWVKQLNSTWRWSVLTVLHQDWLNGKEYKCKVSNKALPA

447 PIEKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN Clone CH2D.3

GQPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH

YTQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSDDWTWVKFNWYVDG

VEVHNAKTKPWIAQPNSTWRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI

448 EKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG Clone CH2D.4

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSDDWEWVKFNWYVDG

VEVHNAKTKPWKLQLNSTWRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP

449 IEKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN Clone CH2D.5

GQPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH

YTQKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPWVWFYWYVDG

450 VEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCSWNIALWWS Clone CH2E3.1 IEKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN GQPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH

YTQKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPWGFRWYVDGV

EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCRVSNSALTWKIE

451 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2E3.2

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPWGFRWYVDGV

EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCRVSNSALSWRIE

452 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2E3.3

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPIVGFRWYVDGVE

VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCRVSNSALRWRIE

453 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ Clone CH2E3.4

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPAVGFEWYVDGV EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCQVFNWALDWVI

454 EKTISKAKGQPP^PQ TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG Clone CH2E3.5 QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV Wild-type human

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Fc sequence

455 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ positions 231-447

PEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT EU index

QKSLSLSPGK numbering

MMDQARSAFSNLFGGEPLSYTRFSLARQVDGDNSHVEMKLAVDEEENADN

NTKANVTKPKRCSGSICYGTIAVIVFFLIGFMIGYLGYCKGVEPKTECERLAG

TESPVP^EPGEDFPAAPJ LYWDDLKRKLSEKLDSTDFTGTIKLLNENSYVPRE

AGSQKDENLALYVENQFREFKLSKVWRDQHFVKIQVKDSAQNSVIIVDKNG

RLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVR

AGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGTGDPYTP

GFPSFNHTQFPPSRSSGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCR Human transferrin

456 MWSESKNVKLTVSNVLKEIKILNIFGVIKGFVEPDHYVVVGAQRDAWGPGA receptor protein 1

AKSGVGTALLLKLAQMFSDMVLKDGFQPSRSIIFASWSAGDFGSVGATEWLE (TFR1)

GYLSSLHLKAFTYINLDKAVLGTSNFKVSASPLLYTLIEKTMQNVKHPVTGQF

LYQDSNWASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTMDTY

KELIERIPELNKVARAAAEVAGQFVIKLTHDVELNLDYERYNSQLLSFVRDLN

QYRADIKEMGLSLQWLYSARGDFFRATSRLTTDFGNAEKTDRFVMKKLNDR

VMRVEYHFLSPYVSPKESPFRHVFWGSGSHTLPALLENLKLRKQNNGAFNET

LFRNQLALATWTIQGAANALSGDVWDIDNEF

457 YxTEWSS CH3C motif

458 TxxExxxxF CH3C motif

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

Fc sequence with

459 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQ

hole mutations

PEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Fc sequence with

460 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQ hole and LALA

PEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYT mutations

QKSLSLSPGK

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV Fc sequence with

461 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE hole and YTE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQ mutations PEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Fc sequence with

462 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQ hole, LALA, and

PEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYT YTE mutations

QKSLSLSPGK

Fc sequence with

463 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNG

knob mutation QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Fc sequence with

464 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNG knob and LALA

QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY mutations

TQKSLSLSPGK

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Fc sequence with

465 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNG knob and YTE QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY mutations TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Fc sequence with

466 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNG knob, LALA, and QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY YTE mutations TQKSLSLSPGK

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.20.1

467 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGT

with knob EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF S CS VMHEALHNHYT

mutation QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.20.1

468 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGT

with knob and EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF VFS CS VMHEALHNHYT LALA mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.20.1

469 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGT with knob and EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF VFS CS VMHEALHNHYT LALAPG QKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.20.1

470 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGT

with knob and EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF VFS CS VMHEALHNHYT YTE mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.20.1

471 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGT with knob, LALA, EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF VFS CS VMHEALHNHYT and YTE QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.20.1

472 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGT with knob, EWS S YKTTPP VLD SD GSFFL YSKLT VTKEEWQQGF VFS CS VMHEALHNHYT LALAPG, and QKSLSLSPGK YTE mutations

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

473 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.20.1 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESFGT EWS S YKTTPP VLD SD GSFFL VSKLT VTKEEWQQGF S CS VMHEALHNHYT with hole QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.20.1

474 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESFGT

with hole and EWS S YKTTPP VLD SD GSFFL VSKLT VTXEEWQQGF VFS CS VMHEALHNHYT LALA mutations QKSLSLSPGK

475 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESFGT with hole and EWS S YKTTPP VLD SD GSFFL VSKLT VTKEEWQQGF VFS CS VMHEALHNHYT LALAPG QKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEWCVYVOVSHEDPEVKENWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.20.1

476 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESFGT

with hole and EWS S YKTTPP VLD SD GSFFL VSKLT VTKEEWQQGF VFS CS VMHEALHNHYT YTE mutations QKSLSLSPGK

APEAAGGPS LFPPK^KBTLYITPJiPEWCVVVDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.20.1

477 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESFGT with hole, LALA, EWS S YKTTPP VLD SD GSFFL VSKLT VTKEEWQQGF VFS CS VMHEALHNHYT and YTE QKSLSLSPGK mutations

APEAAGGPS LFPPK^KBTLYITPJiPEWCVVVDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.20.1

478 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESFGT with hole, EWS S YKTTPP VLD SD GSFFL VSKLT VTKEEWQQGF VFS CS VMHEALHNHYT LALAPG, and QKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEWCVYVDVSHEDPEVKENWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.2

479 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY

mutation TQKSLSLSPGK

APEAAGGPS LFPPKPKDTLMISRTPEVTCVYVOVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.2

480 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob and

TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALA mutations

TQKSLSLSPGK

APEAAGGPS LFPPKPKDTLMISRTPEVTCVYVOVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.2

481 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob and

TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG

TQKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEWCVYVOVSHEDPEVKENWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.2

482 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob and TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY YTE mutations TQKSLSLSPGK

APEAAGGPS LFPPK^KBTLYITPJiPEWCVVVDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.2

483 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob, LALA, TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY and YTE TQKSLSLSPGK mutations

APEAAGGPS LFPPK^KBTLYITPJiPEWCVVVDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.2

484 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob,

TEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG, and

TQKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEWCVYVDVSHEDPEVKENWYVDGV

Clone

485 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.2 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT EWANYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT with hole QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.2

486 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

with hole and EWANYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT LALA mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.2

487 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole and EWANYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG QKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.2

488 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

with hole and

EWANYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT YTE mutations

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.2

489 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole, LALA, EWANYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT and YTE QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.2

490 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole, EWANYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG, and QKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.3

491 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY

mutation TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.3

492 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob and

TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALA mutations

TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.3

493 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob and

TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG

TQKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.3

494 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob and TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY YTE mutations TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.3

495 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob, LALA, TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY and YTE TQKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.3

496 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob,

TEWV YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG, and

TQKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

497 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.3 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT EWVNYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT with hole QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.3

498 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

with hole and EWVNYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT LALA mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.3

499 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole and EWVNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG QKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.3

500 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

with hole and

EWVNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT YTE mutations

QKSLSLSPGK

501 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole, LALA, EWVNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT and YTE QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.3

502 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole, EWVNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG, and QKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.4

503 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY

mutation TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.4

504 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob and

TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY LALA mutations

TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.4

505 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob and

TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY LALAPG

TQKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.4

506 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

with knob and TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY YTE mutations TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.4

507 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob, LALA,

TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY and YTE

TQKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.4

508 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG with knob,

TEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVMHEALHNHY LALAPG, and

TQKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

509 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.4 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT EWSNYKTTPPVLDSDGSFFLVSKLTVSKEEWQQGFVFSCSVMHEALHNHYT with hole QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23.4

510 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

with hole and

EWSNYKTTPPVLDSDGSFFLVSKLTVSKEEWQQGFVFSCSVMHEALHNHYT LALA mutations

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.4

511 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole and

EWSNYKTTPPVLDSDGSFFLVSKLTVSKEEWQQGFVFSCSVMHEALHNHYT LALAPG

QKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.4

512 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

with hole and

EWSNYKTTPPVLDSDGSFFLVSKLTVSKEEWQQGFVFSCSVMHEALHNHYT YTE mutations

QKSLSLSPGK

APEAAGGPS LFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23.4

513 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole, LALA,

EWSNYKTTPPVLDSDGSFFLVSKLTVSKEEWQQGFVFSCSVMHEALHNHYT and YTE

QKSLSLSPGK mutations

APEAAGGPS LFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23.4

514 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT with hole,

EWSNYKTTPPVLDSDGSFFLVSKLTVSKEEWQQGFVFSCSVMHEALHNHYT LALAPG, and

QKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21.17.2

515 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVLWESYG

with knob TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY

mutation TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21.17.2

516 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVLWESYG

with knob and

TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALA mutations

TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.21.17.2

517 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVLWESYG with knob and

TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG

TQKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21.17.2

518 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVLWESYG

with knob and TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY YTE mutations TQKSLSLSPGK

APEAAGGPS LFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21.17.2

519 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVLWESYG with knob, LALA, TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY and YTE TQKSLSLSPGK mutations

APEAAGGPS LFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.21.17.2

520 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVLWESYG with knob,

TEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG, and

TQKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

521 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21.17.2 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVLWESYGT EWASYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT with hole QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.21.17.2

522 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVLWESYGT

with hole and

EWASYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT LALA mutations

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.21.17.2

523 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVLWESYGT with hole and

EWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG

QKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21.17.2

524 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVLWESYGT

with hole and

EWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT YTE mutations

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.21.17.2

525 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVLWESYGT with hole, LALA,

EWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT and YTE

QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.21.17.2

526 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVLWESYGT with hole,

EWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG, and

QKSLSLSPGK YTE mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE Clone

527 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG CH3C.35.23 with TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY knob mutation TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23 with

528 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

knob and LALA

TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY

mutations

TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23 with

529 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG knob and

TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY LALAPG

TQKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLYITPJiPEVTCVVVDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23 with

530 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

knob and YTE TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY

mutations TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23 with

531 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG

knob, LALA, and TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY YTE mutations TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone

EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23 with

532 KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYG knob, LALAPG,

TEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALHNHY and YTE

TQKSLSLSPGK mutations

APELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV Clone

533 EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23 with KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT hole mutations EWSNYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPPJiEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

CH3C.35.23 with

534 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

hole and LALA EWSNYKTTPPVLDSDGSFFLVSKLTVT EEWQQGFVFSCSVMHEALHNHYT

mutations

QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23 with

535 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT hole and EWSNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT LALAPG QKSLSLSPGK mutations

APELLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23 with 536 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

hole and YTE EWSNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT

mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE CH3C.35.23 with 537 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT

hole, LALA, and EWSNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT YTE mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV Clone EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE CH3C.35.23 with 538 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESYGT hole, LALAPG, EWSNYKTTPPVLDSDGSFFLVSKLTVTKEEWQQGFVFSCSVMHEALHNHYT and YTE QKSLSLSPGK mutations

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Fc sequence with EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE

hole and 539 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQ

LALAPG PEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

Fc sequence with EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE

hole, LALAPG, 540 I KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQ

and YTE PEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYT

mutations QKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV

Fc sequence with EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE

knob and 541 I KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNG

LALAPG QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

mutations TQKSLSLSPGK

APEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGV

Fc sequence with EVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALGAPIE

knob, LALAPG, 542 I KTISKAKGQPP^PQ TLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNG

and YTE QPEN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY

mutations TQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2 543 I KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD

with knob and VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALA mutations GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC LVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

2F09-Fd fused to GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

544 CH3C.35.21.17.2

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

with knob and SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD LALAPG VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN mutations GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC LVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

545 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVWD

with knob and

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

YTE mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC

LVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ

GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN 2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

546 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD with knob, LALA,

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN and YTE

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC mutations

LVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ

GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN 2F09-Fd fused to SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

547 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD with knob, VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALAPG, and GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC YTE mutations LVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

548 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

with hole

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

549 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD

with hole and

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALA mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN 2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

550 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD with hole and

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALAPG

GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA mutations

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

551 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVWD

with hole and

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

YTE mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPP^PQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN 2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

552 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD with hole, LALA,

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN and YTE

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA mutations

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN 2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD CH3C.35.21.17.2

553 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD with hole,

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALAPG, and

GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA YTE mutations

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD 2F09-Fd fused to

554 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD Fc with knob

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN mutation

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC

LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ

GNVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD 2F09-Fd fused to

555 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD Fc with knob and

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALA mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC

LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ

GNVFSCSVMHEALHNHYTQKSLSLSPGK

2F09-Fd fused to SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

Fc with knob and

556 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD

LALAPG VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

mutations GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

2F09-Fd fused to

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

557 Fc with knob and

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD YTE mutations

KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVWD

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC

LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ

GNVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD

Fc with knob,

558 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD

LALA, and YTE

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC

LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ

GNVFSCSVMHEALHNHYTQKSLSLSPGK

2F09-Fd fused to SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

Fc with knob,

559 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD

LALAPG, and VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN YTE mutations GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLWC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD 2F09-Fd fused to

560 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD Fc with hole and

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN LALA mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

Fc with hole and

561 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWD

LALAPG

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

mutations

GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD 2F09-Fd fused to

562 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVWD Fc with hole and

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN YTE mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

2F09-Fd fused to

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

Fc with hole,

563 KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD

LALA, and YTE

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

mutations

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

2F09-Fd fused to

564 GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

Fc with hole, GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD LALAPG, and

KKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWD YTE mutations

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCA

VKGFYPSDIAVEWESNGQPEN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

565 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP

with knob and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT LALA mutations

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLWCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

566 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP with knob and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT LALAPG

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT mutations

KNQVSLWCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

567 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLYITREPE

with knob and

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV YTE mutations

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLWCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

568 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP with knob, LALA,

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT and YTE

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT mutations

KNQVSLWCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

569 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP with knob,

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT LALAPG, and

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT YTE mutations

KNQVSLWCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

570 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

with hole

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

mutations

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLSCAVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

571 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP

with hole and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLT

LALA mutations

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLSCAVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

572 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP with hole and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT LALAPG

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT mutations

KNQVSLSCAVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

573 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLYITREPE

with hole and

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV YTE mutations

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLSCAVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

574 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP with hole, LALA,

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT and YTE

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT mutations

KNQVSLSCAVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

575 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP with hole,

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT LALAPG, and

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT YTE mutations

KNQVSLSCAVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLVSKLT

VTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH 1F4-Fd fused to

576 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE Fc with knob

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV mutation

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV

DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

1F4-Fd fused to

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

577 Fc with knob and

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH LALA mutations

KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

Fc with knob and

578 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP

LALAPG

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT

mutations

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH 1F4-Fd fused to

579 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTL YITREPE Fc with knob and

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV YTE mutations

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLWCLVKGFYPSDIAVEWESNGQPEN YKTTPPVLDSDGSFFLYSKLTV

DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

Fc with knob,

580 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP

LALA, and YTE

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT

mutations

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

Fc with knob,

581 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP

LALAPG, and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT YTE mutations

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH 1F4-Fd fused to

582 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP Fc with hole and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT LALA mutations

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

Fc with hole and

583 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP

LALAPG

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT

mutations

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI 1F4-Fd fused to

584 RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI Fc with hole and

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP YTE mutations EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTL YITREPE

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLSCAVKGFYPSDIAVEWESNGQPEN YKTTPPVLDSDGSFFLVSKLTV

DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

Fc with hole,

585 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP

LALA, and YTE

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT

mutations

VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDRFTISRDDSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

1F4-Fd fused to

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

Fc with hole,

586 KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREP

LALAPG, and

EWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT YTE mutations

VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELT

KNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT

VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

587 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

CH3C.35.20.1

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL

VKGFYPSDIAVEWESFGTEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

588 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

CH3C.35.23.2

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL

VKGFYPSDIAVEWESYGTEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ

GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

589 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

CH3C.35.23.3

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL

VKGFYPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQ

GFVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

590 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

CH3C.35.23.4

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL

VKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQG

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

591 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

CH3C.35.21.17.2

VSHEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL

VKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVHVKQSGPELVEPGASVKISCKTSGYTFASYAISWVKQRTGQGLEWIGEIYP

GSGDTYYDEKFKGKATLTADKS S ST AYMELRSLTSED S AVYFCASD YD AYW

GQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVD

2F09-Fd fused to

592 KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD

CH3C.35.23

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL

VKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQG

FVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

1F4-Fd fused to

593 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.20.1

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESFGTEWSSYKTTPPVLDSDGSFFLYSKLTVT

KEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

1F4-Fd fused to

594 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23.2

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWANYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

1F4-Fd fused to

595 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23.3

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

1F4-Fd fused to

596 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23.4

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTV

SKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSNNYATYYADSVKDRFTISRDDSESKFHLQMNNLKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP 1F4-Fd fused to

597

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH CH3C.35.21.17.2

KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTV TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFSFNTYAMNWVRQAPGKGLEWVARI

RmSN YATYYADSVKDPJTISPJ^DSESKFHLQMN LKTEDTAIYYCVRQGI

DFYAMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

1F4-Fd fused to

598 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

RSKISNYATYYADSVKDPJTISPJ^DSQSMLYLQMN LKTEDTAMYYCVTGS

SYGWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

46D2-Fd fused to

599 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.20.1

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESFGTEWSSYKTTPPVLDSDGSFFLYSKLTVT

KEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

RSKISNYATYYADSVKDRFTISRDDSQSMLYLQMNNLKTEDTAMYYCVTGS

SYGWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

46D2-Fd fused to

600 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23.2

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWANYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

RSKISNYATYYADSVKDRFTISRDDSQSMLYLQMNNLKTEDTAMYYCVTGS

SYGWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

46D2-Fd fused to

601 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23.3

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

RSKISNYATYYADSVKDRFTISRDDSQSMLYLQMNNLKTEDTAMYYCVTGS

SYGWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP

EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

46D2-Fd fused to

602 KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

CH3C.35.23.4

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTV

SKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

RSKISNYATYYADSVKDRFTISRDDSQSMLYLQMNNLKTEDTAMYYCVTGS

SYGWFAYWGQGTLVTVSASASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF

PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVN

46D2-Fd fused to

603 HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT

CH3C.35.21.17.2

PEWCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVL

TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEL

TKNQVSLTCLVKGFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKL

TVTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMHWVRQAPGKGLEWVARI

46D2-Fd fused to

604 RSKISNYATYYADSVKDRFTISRDDSQSMLYLQMNNLKTEDTAMYYCVTGS

CH3C.35.23

SYGWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNH

KP SNTKVDKKVEPKS CDKTHTCPPCP APELLGGP S VFLFPPKPKDTLMISRTPE

WCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTV

LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK

NQVSLTCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTV

TKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGFIS

SDGNSDYNPSLKNRISVTRDTSKNQFFLQLNSVTTEDTATYYCLRGDVDWGQ

GTTLTVSSAST GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA

LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT VDKKV

2D02-Fd fused to

605 EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHE

CH3C.35.20.1

DPEVKFNWYVDGVEVHNAKmPPJiEQYNSTYRWSVLTVLHQDWLNGKEY

KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF

YPSDIAVEWESFGTEWSSYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSC

SVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGFIS

SDGNSDYNPSLKNRISVTRDTSKNQFFLQLNSVTTEDTATYYCLRGDVDWGQ

GTTLTVSSAST GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA

LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT VDKKV

2D02-Fd fused to

606 EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHE

CH3C.35.23.2

DPEVKFNWYVDGVEVHNAKT PPJiEQYNSTYRVVSVLTVLHQDWLNGKEY

KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF

YPSDIAVEWESYGTEWANYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFS

CSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGFIS

SDGNSDYNPSLKNRISVTRDTSKNQFFLQLNSVTTEDTATYYCLRGDVDWGQ

GTTLTVSSAST GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA

LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT VDKKV

2D02-Fd fused to

607 EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHE

CH3C.35.23.3

DPEVKFNWYVDGVEVHNAKmPPJiEQYNSTYRWSVLTVLHQDWLNGKEY

KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF

YPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFS

CSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGFIS

SDGNSDYNPSLKNRISVTRDTSKNQFFLQLNSVTTEDTATYYCLRGDVDWGQ

GTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA

LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV

2D02-Fd fused to

608 EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHE

CH3C.35.23.4

DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY

KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF

YPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFS

CSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGFIS

SDGNSDYNPSLKNRISVTRDTSKNQFFLQLNSVTTEDTATYYCLRGDVDWGQ

GTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA

LTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKKV

2D02-Fd fused to

609 EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHE

CH3C.35.21.17.2

DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY

KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF

YP SDI AVLWES YGTEW AS YKTTPP VLD SDGSFFL YSKLTVTKEEWQQGF VF S

CSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGFIS

SDGNSDYNPSLKNRISVTRDTSKNQFFLQLNSVTTEDTATYYCLRGDVDWGQ

GTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA

LTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKKV

2D02-Fd fused to

610 EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHE

CH3C.35.23

DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY

KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF

YPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFS

CSVMHEALHNHYTQKSLSLSPGK EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

DDGRNNY PSLKNRISITRDTSKNQLFLKLNSVTTEDTATYYCVRGDYGPWG

QGTTVTVSSAST GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFPAVLQSSGLYSLSSVW SSSLGTQTYICNVNHKPSNT VDKK

2F10-Fd fused to

611 VEPKSCDKTHTCPPCPAPELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVS

CH3C.35.20.1

HEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK

GFYPSDIAVEWESFGTEWSSYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFVF

SCSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

DDGRNNYNPSLKNRISITRDTSKNQLFLKLNSVTTEDTATYYCVRGDYGPWG

QGTTVTVSSAST GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFPAVLQSSGLYSLSSVW SSSLGTQTYICNVNHKPSNT VDKK

2F10-Fd fused to

612 VEPKSCDKTHTCPPCPAPELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVS

CH3C.35.23.2

HEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK

GFYPSDIAVEWESYGTEWANYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFV

FSCSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

DDGRNNYNPSLKNRISITRDTSKNQLFLKLNSVTTEDTATYYCVRGDYGPWG

QGTTVTVSSAST GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFPAVLQSSGLYSLSSVW SSSLGTQTYICNVNHKPSNT VDKK

2F10-Fd fused to

613 VEPKSCDKTHTCPPCPAPELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVS

CH3C.35.23.3

HEDPEVKFNWYVDGVEVHNAKTKPP^EQYNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK

GFYPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFFLYSKLTVT EEWQQGFV

FSCSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

DDGRNNYNPSLKNRISITRDTSKNQLFLKLNSVTTEDTATYYCVRGDYGPWG

QGTTWVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFPAVLQSSGLYSLSSVW SSSLGTQTYICNVNHKPSNTKVDKK

2F10-Fd fused to

614 VEPKSCDKTHTCPPCPAPELLGGPS LFPPKPKDTLMISRTPEVTCVVVDVS

CH3C.35.23.4

HEDPEVKFN VDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGK

EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK

GFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFV

FSCSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

DDGRNNYNPSLKNRISITRDTSKNQLFLKLNSVTTEDTATYYCVRGDYGPWG

QGTTWVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK

2F10-Fd fused to

615 VEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS

CH3C.35.21.17.2

HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGK

EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK

GFYPSDIAVLWESYGTEWASYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFV

FSCSVMHEALHNHYTQKSLSLSPGK

EVKLQESGPGLVKPSQSLSLTCSVTGYSITSGYYWNWIRQFPGNKLEWLGYIS

DDGRNNYNPSLKNRISITRDTSKNQLFLKLNSVTTEDTATYYCVRGDYGPWG

QGTTWVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK

2F10-Fd fused to

616 VEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS

CH3C.35.23

HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGK

EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK

GFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFV

FSCSVMHEALHNHYTQKSLSLSPGK

DIVLTQTPLSLPVSLGDQASISCRSSQSLVHRNGNTYLHWYLQKPGQSPKLLI YRISNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPLTFGAGT

617 KLEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLN FYPREAKVQWKVDNAL 2F09 light chain QSGNSQES VTEQD SKD STYSL S STLTL SKAD YEKHKVYACEVTHQGLS SP VT KSFNRGEC DVLMTQSPLSLSVSLGDQASISCRSSRDIVHSNGNTYLEWYLQKPGQSPKLLI

YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGTLIPLTFGAG

618 TKLELKRTVAAPSVFIFPPSDEQLKSGTASWCLLN FYPREAKVQWKVDNA 1F4 light chain LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC

QIVLTRSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSK

LASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGAGTKLEL

619 KRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGN 46D2 light chain

SQESWEQDSKDSTYSLSSTLTLSKADYEKHK ACEVTHQGLSSPVT SFNR

GEC

DVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGETYLNWLLQRPGQSPKRLI

YLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGG

620 GTKLELKRTVAAPSVFIFPPSDEQLKSGTASWCLLN FYPREAKVQWKVDN 2D02 light chain

ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP

VTKSFNRGEC

DVVMTQTPLTLSVTIGQPAFISCKSNQSLLDYDGETYLNWLFLRPGQSPKRLI

YLVSKQDSGVPDRFTGIGSGTDFTLKINRVEAEDLGIYYCWQGTHFPQTFGG

621 GTKLELKRTVAAPSVFIFPPSDEQLKSGTASWCLLN FYPREAKVQWKVDN 2F10 light chain

ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP

VTKSFNRGEC

CDR-H1

622 GFXiFSDYGMH, wherein Xi is I or T consensus sequence

CDR-H2

623 ISSGNX₁TVYYADTVKX₂, wherein Xi is N or D and X₂ is G or D consensus sequence

CDR-H1

X1YX2IX3X4X5X6Y, wherein Xi is G or D; X₂ is P or S; X₃ is T or P; X₄ is S, R, or

624 consensus N; X₅ is G, Y, N, or F; and X₆ is Y or F

sequence

CDR-H1

625 GYSITxxYY consensus sequence

CDR-H1

626 GX₁TFTDYYX₂N, wherein Xi is H, Y, F, or N and X₂ is M or I consensus sequence

IX1X2DGX3X4X5YX6PSLX7X8, wherein Xi is G, N, S, or T; X₂ is D, Y, S, Q, or T; CDR-H2

627 X₃ is S, Y, N, or T; X₄ is N, S, D, T, or H; X₅ is N, D, or S; X₆ is N, T, or S; X₇ is K consensus or E; and Xs is N or D sequence

CDR-H2

628 IxxDGxxxYxPSLKN consensus sequence

IXiPX₂X3GX4X₅X6X7X8QX9FXioXii, wherein X₁ is H or N; X₂ is Y, N, or H; X₃ is CDR-H2

629 T, S, R, or N; X₄ is D, E, or G; X₅ is T, A, or I; X₆ is R, N, Q, or S; X₇ is Y or N; X₈ consensus is N, S, T, or A; X₉ is R, K, or N; X_i0 is R or K; and Xn is G or D sequence

CDR-H2

630 IHPYxGxxxxxQxFKG consensus sequence

CDR-H3

X1X2GDX3X4, wherein Xi is A, I, L, T, or V; X₂ is R, S, K, or T; X₃ is N, V, S, L, D,

631 consensus G, or I; and X₄ is Y, V, N, D, H, L, or A

sequence

CDR-H3

632 xxGDVx consensus sequence

CDR-L1

X₁X₂X₃X₄SLLX₅X₆DGX₇TYLN, wherein Xi is K or R; X₂ is S or T; X₃ is S or N; X₄

633 consensus is Q, R, K, or L; X₅ is D or Y; X₆ is S, H, I, R, Y, N, or K; and X₇ is E or K

sequence

CDR-L1

634 KSSxSLLxxDGETYLN consensus sequence CDR-L2

X1SX2X3X4S, wherein Xi is V, T, or A; X₂ is K, Y, or Q; X₃ is L or R; and X₄ is D,

635 consensus A, or Y

sequence

CDR-L3

X₁QX₂X₃X₄X₅PX₆T, wherein Xi is W, Q, or M; X₂ is G, W, Y, or H; X₃ is T, S, N, or

636 consensus L; X₄ is H, S, or E; X₅ if F, N, or Y; X₆ is Q, P, L, R, or F

sequence

CDR-H1

X₁YX₂ITX₃X₄X₅YWN, wherein Xi is G or D; X₂ is S or P; X₃ is S, N, or R; X₄ is G,

637 consensus

sequence

CDR-H1

638 GX₁TFTDYYX₂N, wherein Xi is H, Y, F, or N and X₂ is M or I consensus sequence

CDR-H1

639 XiYSITSGYY, wherein X₁ is D or G consensus sequence

CDR-H2

640 ISDDGRNXiYNPSLKN, wherein Xi is D or N consensus sequence

CDR-H3

641 X1RGDX2GP, wherein Xi is L or V and X₂ is N or Y consensus sequence

CDR-L1

KSX₁QSLX₂DX₃DGX₄TYLN, wherein Xi is S or N; X₂ is L or F; X₃ is S or Y; and

642 consensus sequence

CDR-L2

643 VSKXiDS wherein Xi is L or Q consensus sequence

CDR-H1

644 XiYSITSGYYWN, wherein X₁ is D or G consensus sequence

CDR-L2

645 LVSKXiDS, wherein Xi is L or Q consensus sequence

CDR-H2

646 ISRGSSAIYYADAVXiG, wherein Xi is K or R consensus sequence

CDR-H1

647 GYSITSGXiYWN, wherein Xi is H or Y consensus sequence

CDR-H1

648 GFX1FNTYAMX2, wherein Xi is T or S and X₂ is H or N consensus sequence

CDR-H2

RIRX₁KX₂X₃NYATYYADSVDK, wherein Xi is S or T; X₂ is S or I; and X₃ is N or

649 consensus S

sequence

CDR-H2

X₁X₂X₃X₄DGX₅X₆NYNPSLKN, wherein Xi is F or Y; X₂ is I or L; X₃ is S or T; X₄

650 consensus is D or Y; X₅ is I or N; and X₆ is N or K

sequence

X₁SX₂X₃X₄X₅X₆X₇X₈X₉GX₁₀TYLX₁₁, wherein X₁ is K or R; X₂ is S or G; X₃ is Q or CDR-L1

651 R; X₄ is S or D; X₅ is I or L; X₆ is I, V, or L; X₇ is D or H; X₈ is S or V; X₉ is D, Y, or consensus N; X₁₀ is E, S, or N; and Xn is N or E sequence

CDR-L2

X1X2SX3X4X5S, wherein Xi is K, D, or L; X₂ is V or T; X₃ is N, K, or T; X₄ is R or

652 consensus

sequence

CDR-L3

X1QX2X3X4X5PX6T, wherein Xi is Q, W, or F; X₂ is W or G; X₃ is S or T; X₄ is S, H,

653 consensus R, or L; X₅ is N, F, V, G, or I; and X₆ is P, Q, or L

sequence

Claims

WHAT IS CLAIMED IS: 1. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 106-125 of SEQ ID NO: 1.

2. The isolated antibody of claim 1, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 106-120 or residues 111-125 of SEQ ID NCv l .

3. The isolated antibody of claim 1, comprising one or more complementarity determining regions (CDRs) selected from the group consisting of:

(a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240;

(b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:81, 82, and 244;

(c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:205 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:205;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and

(f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:231 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO :231.

4. The isolated antibody of claim 1, comprising one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:62, 63, and 240;

(b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 81, 82, and 244;

(c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 121, 267, and 268;

(d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205;

(e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and

(f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

5. The isolated antibody of claim 4, comprising:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 62;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81; and

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 121.

6. The isolated antibody of claim 4, comprising:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 62;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81; and (c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:267.

7. The isolated antibody of claim 4, comprising:

(a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205;

(b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and (c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

8. The isolated antibody of claim 4, comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:62, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 121, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

9. The isolated antibody of claim 4, comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:62, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:81, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:267, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:205, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:231.

10. The isolated antibody of claim 1, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:2-3 and 266.

11. The isolated antibody of claim 1, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 146- 148.

12. The isolated antibody of claim 1, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:2-3 and 266, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 146-148.

13. The isolated antibody of claim 1, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:2 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO : 147.

14. The isolated antibody of claim 1, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:266 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO : 147.

15. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 71-105 of SEQ ID NO: 1.

16. The isolated antibody of claim 15, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 71-85, residues 86-100, or residues 91-105 of SEQ ID NO: 1.

17. The isolated antibody of claim 15, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:64-78, 241, 242, 317, and 318 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:64- 78, 241, 242, 317, and 318;

(b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:83-115, 245, and 246 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:83-l 15, 245, and 246;

(c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:206-220 and 251 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:206-220 and 251 ;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:224-228 and 253 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:224-228 and 253; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:232-237 and 255 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:232-237 and 255.

18. The isolated antibody of claim 15, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 64-78, 241, 242, 317, and 318;

(b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:83-115, 245, and 246;

(c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 122-141 and 249;

(d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206-220 and 251;

(e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:224-228 and 253; and

(f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232-237 and 255.

19. The isolated antibody of claim 18, comprising:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:65;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:88; and

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 126.

20. The isolated antibody of claim 18, comprising:

(a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:218;

(b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228; and

(c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

21. The isolated antibody of claim 18, comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:88, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 126, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:218, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

22. The isolated antibody of claim 15, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:4-55.

23. The isolated antibody of claim 15, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 149-197.

24. The isolated antibody of claim 15, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:4-55, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 149-197.

25. The isolated antibody of claim 15, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 12 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 157.

26. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 66-85 of SEQ ID NO: 1.

27. The isolated antibody of claim 26, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 66-80 or residues 71-85 of SEQ ID NO: l .

28. The isolated antibody of claim 26, comprising one or more CDRs selected from the group consisting of: (a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:79 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 79;

(b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 116 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 116;

(c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 142 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 142;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:221;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:229 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:229; and

(f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:238 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO :238.

29. The isolated antibody of claim 26, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:79;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116;

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142;

(d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:221;

(e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229; and

(f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

30. The isolated antibody of claim 29, comprising:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:79;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116; and

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142.

31. The isolated antibody of claim 29, comprising:

(a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:221;

(b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229; and

(c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

32. The isolated antibody of claim 29, comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:79, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 116, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 142, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:221, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:229, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:238.

33. The isolated antibody of claim 26, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:56.

34. The isolated antibody of claim 26, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 198.

35. The isolated antibody of claim 26, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:56, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO : 198.

36. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 81-115 of SEQ ID NO: l .

37. The isolated antibody of claim 36, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 81-95, residues 86-100, residues 91-105, or residues 101-115 of SEQ ID NCv l .

38. The isolated antibody of claim 36, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319;

(b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 117, 118, and 247 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 117, 118, and 247;

(c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320; and

(f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:237 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:237.

39. The isolated antibody of claim 36, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:65, 80, 243, and 319;

(b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 117, 118, and 247;

(c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 143, 144, and 250;

(d) a light chain CDRl comprising the amino acid sequence of any one of SEQ ID NOs:218, 219, 222, and 252;

(e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:228, 230, 254, and 320; and

(f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

40. The isolated antibody of claim 39, comprising:

(a) a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:65;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 118; and

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 144.

41. The isolated antibody of claim 39, comprising:

(a) a light chain CDRl comprising the amino acid sequence of SEQ ID NO:219;

(b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:230; and

(c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

42. The isolated antibody of claim 39, comprising a heavy chain CDRl comprising the amino acid sequence of SEQ ID NO:65, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 118, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 144, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:219, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:230, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:237.

43. The isolated antibody of claim 36, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:57-58.

44. The isolated antibody of claim 36, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 199-201.

45. The isolated antibody of claim 36, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:57-58, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 199-201.

46. The isolated antibody of claim 36, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:58 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO :201.

47. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 36-50 and residues 86-115 of SEQ ID NO: l .

48. The isolated antibody of claim 47, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 36-50, residues 86-100, residues 96-110, or residues 101-115 of SEQ ID NO: l .

49. The isolated antibody of claim 47, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:63 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 63; (b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 119, 120, and 248 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 119, 120, and 248;

(c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 145 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 145;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:220;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:228 or 316 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:228 or 316; and

(f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:239 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:239.

50. The isolated antibody of claim 47, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:63;

(b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 119, 120, and 248;

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145;

(d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:220;

(e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228 or 316; and

(f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

51. The isolated antibody of claim 50, comprising: (a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:63;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 119; and

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145.

52. The isolated antibody of claim 50, comprising:

(a) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:220;

(b) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228; and

(c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

53. The isolated antibody of claim 50, comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:63, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 119, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 145, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:220, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:228, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:239.

54. The isolated antibody of claim 47, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:59-61.

55. The isolated antibody of claim 47, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:202-204.

56. The isolated antibody of claim 47, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:59-61, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:202-204.

57. The isolated antibody of claim 47, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:60 and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO :203.

58. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 86-105 or residues 121-140 of SEQ ID NO: 1.

59. The isolated antibody of claim 58, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 86-100, residues 91-105, residues 121-135, or residues 126-140 of SEQ ID NO: 1.

60. The isolated antibody of claim 58, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 86-105 of SEQ ID NO: 1.

61. The isolated antibody of claim 58, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 121-140 of SEQ ID NO: 1.

62. The isolated antibody of claim 58, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 126-140 of SEQ ID NO: 1.

63. The isolated antibody of claim 58, wherein the antibody or antigen- binding portion thereof recognizes an epitope within residues 86-105 and residues 121-140 of SEQ ID NO: l .

64. The isolated antibody of claim 58, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310;

(b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 311-312 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 311-312; (c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314; and

(f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315 or having up to two amino acid substitutions relative to the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315.

65. The isolated antibody of claim 58, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:65, 276-278, and 309-310;

(b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:279, 281-284, and 311-312;

(c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:285-288 and 290-291;

(d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs:206, 218, 301-304, and 313;

(e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs:223, 224, 228, 305, and 314; and

(f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs:232, 237, 306-308, and 315.

66. The isolated antibody of claim 58, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:269-272 and 274-275.

67. The isolated antibody of claim 58, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:292-295 and 298-300.

68. The isolated antibody of claim 58, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:269-272 and 274-275, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:292-295 and 298-300.

69. An isolated antibody or antigen-binding portion thereof that specifically binds to a human alpha-synuclein protein and recognizes an epitope within residues 61-140 of SEQ ID NO: 1.

70. The isolated antibody of claim 69, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:277 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:277;

(b) a heavy chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:280 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:280;

(c) a heavy chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:289 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:289;

(d) a light chain CDR1 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:304 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 304;

(e) a light chain CDR2 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:223; and (f) a light chain CDR3 having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:308 or having up to two amino acid substitutions relative to the amino acid sequence of SEQ ID NO:308.

71. The isolated antibody of claim 69, comprising one or more CDRs selected from the group consisting of:

(a) a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:277;

(b) a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:280;

(c) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:289;

(d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:304;

(e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:223; and

(f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:308.

72. The isolated antibody of claim 69, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:273.

73. The isolated antibody of claim 69, comprising a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:296-297.

74. The isolated antibody of claim 69, comprising a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:273, and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs:296-297.

75. The isolated antibody of any one of claims 1 to 74, wherein the antibody comprises a first Fc polypeptide and optionally a second Fc polypeptide.

76. The isolated antibody of claim 75, wherein the antibody comprises the first Fc polypeptide and the second Fc polypeptide.

77. The isolated antibody of claim 75 or 76, wherein the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide.

78. The isolated antibody of any one of claims 1 to 74, wherein the antibody comprises:

(a) a first antigen-binding portion comprising a first variable region that specifically binds to the human alpha-synuclein protein, wherein the first antigen-binding portion comprises (i) a first heavy chain comprising a first Fc polypeptide and (ii) a first light chain; and

(b) a second antigen-binding portion comprising a second variable region that specifically binds to the human alpha-synuclein protein, wherein the second antigen- binding portion comprises (i) a second heavy chain comprising a second Fc polypeptide and (ii) a second light chain;

wherein the first Fc polypeptide and the second Fc polypeptide form an Fc dimer.

79. The isolated antibody of claim 78, wherein the first Fc polypeptide is a modified Fc polypeptide and/or the second Fc polypeptide is a modified Fc polypeptide.

80. The isolated antibody of claim 78 or 79, wherein the first variable region and the second variable region recognize the same epitope in the human alpha-synuclein protein.

81. The isolated antibody of claim 78 or 79, wherein the first variable region and the second variable region recognize different epitopes in the human alpha-synuclein protein.

82. The isolated antibody of any one of claims 77 to 81, wherein the first Fc polypeptide and the second Fc polypeptide each contain modifications that promote heterodimerization.

83. The isolated antibody of claim 82, wherein one of the Fc polypeptides has a T366W substitution and the other Fc polypeptide has T366S, L368A, and Y407V substitutions, according to EU numbering.

84. The isolated antibody of any one of claims 77 to 83, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises a native FcRn binding site.

85. The isolated antibody of any one of claims 77 to 83, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises a modification that alters FcRn binding.

86. The isolated antibody of any one of claims 77 to 85, wherein the first Fc polypeptide and the second Fc polypeptide do not have effector function.

87. The isolated antibody of any one of claims 77 to 85, wherein the first Fc polypeptide and/or the second Fc polypeptide includes a modification that reduces effector function.

88. The isolated antibody of claim 87, wherein the modification that reduces effector function comprises the substitutions of Ala at position 234 and Ala at position 235, according to EU numbering.

89. The isolated antibody of any one of claims 77 to 88, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises amino acid changes relative to the native Fc sequence that extend serum half-life.

90. The isolated antibody of claim 89, wherein the amino acid changes comprise substitutions of Tyr at position 252, Thr at position 254, and Glu at position 256, according to EU numbering.

91. The isolated antibody of claim 89, wherein the amino acid changes comprise substitutions of Leu at position 428 and Ser at position 434, according to EU numbering.

92. The isolated antibody of claim 89, wherein the amino acid changes comprise a substitution of Ser or Ala at position 434, according to EU numbering.

93. The isolated antibody of any one of claims 77 to 92, wherein the first Fc polypeptide and/or the second Fc polypeptide specifically binds to a transferrin receptor.

94. The isolated antibody of claim 93, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises at least two substitutions at positions selected from the group consisting of 384, 386, 387, 388, 389, 390, 413, 416, and 421, according to EU numbering.

95. The isolated antibody of claim 94, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises substitutions at least three, four, five, six, seven, eight, or nine of the positions.

96. The isolated antibody of claim 94 or 95, wherein the first Fc polypeptide and/or the second Fc polypeptide further comprises one, two, three, or four substitutions at positions comprising 380, 391, 392, and 415, according to EU numbering.

97. The isolated antibody of any one of claims 94 to 96, wherein the first Fc polypeptide and/or the second Fc polypeptide further comprises one, two, or three substitutions at positions comprising 414, 424, and 426, according to EU numbering.

98. The isolated antibody of any one of claims 94 to 97, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises Trp at position 388.

99. The isolated antibody of any one of claims 94 to 98, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises an aromatic amino acid at position 421.

100. The isolated antibody of claim 99, wherein the aromatic amino acid at position 421 is Trp or Phe.

101. The isolated antibody of any one of claims 94 to 100, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises at least one position selected from the following: position 380 is Trp, Leu, or Glu; position 384 is Tyr or Phe; position 386 is Thr; position 387 is Glu; position 388 is Trp; position 389 is Ser, Ala, Val, or Asn; position 390 is Ser or Asn; position 413 is Thr or Ser; position 415 is Glu or Ser; position 416 is Glu; and position 421 is Phe.

102. The isolated antibody of claim 101, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 positions selected from the following: position 380 is Tip, Leu, or Glu; position 384 is Tyr or Phe; position 386 is Thr; position 387 is Glu; position 388 is Trp; position 389 is Ser, Ala, Val, or Asn; position 390 is Ser or Asn; position 413 is Thr or Ser; position 415 is Glu or Ser; position 416 is Glu; and position 421 is Phe.

103. The isolated antibody of claim 102, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises 11 positions as follows: position 380 is Trp, Leu, or Glu; position 384 is Tyr or Phe; position 386 is Thr; position 387 is Glu; position 388 is Trp; position 389 is Ser, Ala, Val, or Asn; position 390 is Ser or Asn; position 413 is Thr or Ser; position 415 is Glu or Ser; position 416 is Glu; and position 421 is Phe.

104. The isolated antibody of claim 102 or 103, wherein the first Fc polypeptide and/or the second Fc polypeptide has a CH3 domain with at least 85% identity, at least 90% identity, or at least 95% identity to amino acids 111-217 of any one of SEQ ID NOs:336-372 and 467-538.

105. The isolated antibody of claim 102 or 103, wherein the first Fc polypeptide and/or the second Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs:467-538.

106. The isolated antibody of claim 104 or 105, wherein the residues at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 of the positions corresponding to EU index positions 380, 384, 386, 387, 388, 389, 390, 391, 392, 413, 414, 415, 416, 421, 424 and 426 of any one of SEQ ID NOs:336-372 and 467-538 are not deleted or substituted.

107. The isolated antibody of any one of claims 93 to 106, wherein the first Fc polypeptide and/or the second Fc polypeptide binds to the apical domain of the transferrin receptor.

108. The isolated antibody of claim 107, wherein the binding of the antibody or antigen-binding portion thereof to the transferrin receptor does not substantially inhibit binding of transferrin to the transferrin receptor.

109. The isolated antibody of any one of claims 75 to 108, wherein the first Fc polypeptide and/or the second Fc polypeptide has an amino acid sequence identity of at least 75%, or at least 80%, 90%, 92%, or 95%, as compared to the corresponding wild-type Fc polypeptide.

110. The isolated antibody of claim 109, wherein the corresponding wild-type Fc polypeptide is a human IgGl, IgG2, IgG3, or IgG4 Fc polypeptide.

111. The isolated antibody of any one of claims 93 to 110, wherein uptake into the brain of the antibody or antigen-binding portion thereof is at least ten-fold greater as compared to the uptake of the antibody or antigen-binding portion thereof without the modifications in the first Fc polypeptide and/or the second Fc polypeptide that result in transferrin receptor binding.

112. The isolated antibody of any one of claims 75 to 111, wherein one of the Fc polypeptides is not modified to bind to a blood-brain barrier receptor and the other Fc polypeptide is modified to specifically bind to a transferrin receptor.

113. The isolated antibody of any one of claims 1 to 112, wherein the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 20 nM.

114. The isolated antibody of claim 113, wherein the antibody or antigen- binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 20 nM.

115. The isolated antibody of any one of claims 1 to 112, wherein the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 10 nM.

116. The isolated antibody of claim 115, wherein the antibody or antigen- binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 10 nM.

117. The isolated antibody of any one of claims 1 to 112, wherein the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 5 nM.

118. The isolated antibody of claim 117, wherein the antibody or antigen- binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 5 nM.

119. The isolated antibody of any one of claims 1 to 112, wherein the antibody or antigen-binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of less than about 1 nM.

120. The isolated antibody of claim 119, wherein the antibody or antigen- binding portion thereof specifically binds to the human alpha-synuclein protein with a binding affinity of about 1 pM to about 1 nM.

121. The isolated antibody of any one of claims 1 to 120, wherein the antibody or antigen-binding portion thereof specifically binds to monomeric and/or oligomeric human alpha-synuclein protein.

122. The isolated antibody of claim 121, wherein the antibody or antigen- binding portion thereof specifically binds to monomeric and oligomeric human alpha- synuclein protein.

123. The isolated antibody of claim 121, wherein the antibody or antigen- binding portion thereof has a higher binding affinity for oligomeric human alpha-synuclein protein than for monomeric human alpha-synuclein protein.

124. The isolated antibody of claim 121, wherein the antibody or antigen- binding portion thereof has a higher binding affinity for monomeric human alpha-synuclein protein than for oligomeric human alpha-synuclein protein.

125. The isolated antibody of any one of claims 1 to 120, wherein the antibody or antigen-binding portion thereof specifically binds to soluble human alpha- synuclein protein and/or human alpha-synuclein protein fibrils.

126. The isolated antibody of claim 125, wherein the antibody or antigen- binding portion thereof specifically binds to soluble human alpha-synuclein protein and human alpha-synuclein protein fibrils.

127. The isolated antibody of any one of claims 1 to 120, wherein the antibody is a pan-alpha-synuclein antibody or antigen-binding portion thereof.

128. The isolated antibody of claim 127, wherein the pan-alpha-synuclein antibody or antigen-binding portion thereof specifically binds to monomeric human alpha- synuclein protein, oligomeric human alpha-synuclein protein, soluble human alpha-synuclein protein, and human alpha-synuclein protein fibrils.

129. The isolated antibody of any one of claims 1 to 128, wherein the antibody is a monoclonal antibody.

130. The isolated antibody of any one of claims 1 to 128, wherein the antibody is a chimeric antibody.

131. The isolated antibody of any one of claims 1 to 128, wherein the antibody is a humanized antibody.

132. The isolated antibody of any one of claims 1 to 128, wherein the antibody is a fully human antibody.

133. The isolated antibody of any one of claims 1 to 132, wherein the antigen-binding portion is a Fab, a F(ab')₂, a scFv, or a bivalent scFv.

134. A pharmaceutical composition comprising the isolated antibody of any one of claims 1 to 133 and a pharmaceutically acceptable carrier.

135. A bispecific antibody comprising the antibody of any one of claims 1 to 133.

136. The bispecific antibody of claim 135, wherein the bispecific antibody recognizes two different alpha-synuclein epitopes.

137. An isolated polynucleotide comprising a nucleotide sequence encoding the isolated antibody of any one of claims 1 to 133.

138. A vector comprising the polynucleotide of claim 137.

139. A host cell comprising the polynucleotide of claim 137 or the vector of claim 138.

140. An antibody that competes with the isolated antibody of any one of claims 1 to 133 for binding to the human alpha-synuclein protein.

141. A hybridoma cell line that secretes the antibody of any one of claims 58 to 68, wherein the hybridoma cell line is selected from the group consisting of 3D5, 4F5, 46D2, 28B6, 3E12, 1F4, 1G4, and 16D4.

142. An antibody secreted by the hybridoma cell line of claim 141.

143. A method for neutralizing human alpha-synuclein protein in a brain of a subject, the method comprising administering to the subject the isolated antibody of any one of claims 1 to 133, the pharmaceutical composition of claim 134, or the bispecific antibody of claim 135 or 136.

144. The method of claim 143, wherein neutralizing comprises preventing or reducing human alpha-synuclein protein oligomerization and/or aggregation in the brain of the subject.

145. The method of claim 143, wherein neutralizing comprises preventing or reducing the spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject.

146. The method of claim 145, wherein the isolated antibody, the pharmaceutical composition, or the bispecific antibody is administered prior to the broad spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subj ect.

147. The method of claim 146, wherein administering the isolated antibody, the pharmaceutical composition, or the bispecific antibody prior to the broad spreading of human alpha-synuclein protein oligomers and/or fibrils in the brain of the subject increases the ability of the isolated antibody, the pharmaceutical composition, or the bispecific antibody to treat a neurodegenerative disease.

148. The method of any one of claims 143 to 147, wherein the isolated antibody, the pharmaceutical composition, or the bispecific antibody neutralizes monomeric human alpha-synuclein protein, oligomeric human alpha-synuclein protein, soluble human alpha-synuclein protein, human alpha-synuclein protein fibrils, or a combination thereof.

149. The method of claim 148, wherein the isolated antibody or bispecific antibody is a pan-alpha-synuclein antibody or a pan-alpha-synuclein bispecific antibody.

150. The method of any one of claims 143 to 149, wherein the subject has a neurodegenerative disease.

151. The method of claim 150, wherein the neurodegenerative disease is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Huntington's disease, Alzheimer's disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down's syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden-Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle-predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick's disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia.

152. The method of claim 150 or 151, wherein the neurodegenerative disease is a synucleinopathy.

153. The method of claim 152, wherein the synucleinopathy is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy.

154. The method of any one of claims 150 to 153, wherein the isolated antibody, the pharmaceutical composition, or the bispecific antibody is administered during an early stage of the neurodegenerative disease.

155. The method of any one of claims 150 to 154, wherein the subject has not been treated for the neurodegenerative disease, has not been treated with a dopaminergic replacement therapy, or has not been treated with a dopamine agonist therapy.

156. The method of any one of claims 143 to 155, wherein the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is necessary to neutralize the human alpha-synuclein protein in the brain of the subj ect is determined based on an amount of total human alpha-synuclein protein in a sample and/or an amount of human alpha-synuclein protein in a sample that is bound to the isolated antibody or bispecific antibody.

157. The method of claim 156, wherein the sample is obtained from the subject.

158. The method of claim 156 or 157, wherein the sample is selected from the group consisting of an interstitial brain fluid sample, a cerebrospinal fluid (CSF) sample, a blood sample, and a combination thereof.

159. The method of any one of claims 143 to 158, wherein the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is necessary to neutralize the human alpha-synuclein protein in the brain of the subj ect is determined based on a level of neurodegenerative disease progression in the subject and/or an amount of human alpha-synuclein protein spreading in the brain of the subject.

160. The method of claim 159, wherein an imaging method is used to determine the level of neurodegenerative disease progression in the subject and/or the amount of human alpha-synuclein protein spreading in the brain of the subject.

161. The method of claim 160, wherein the imaging method is selected from the group consisting of positron emission tomography (PET) imaging, dopamine transporter (DaT) imaging, magnetic resonance imaging (MRI), and computed tomography (CT) imaging.

162. The method of claim 161, wherein PET imaging comprises using a PET probe that comprises a diabody of the antibody of any one of claims 1 to 133.

163. A method for treating a neurodegenerative disease in a subject, the method comprising administering to the subject the isolated antibody of any one of claims 1 to 133, the pharmaceutical composition of claim 134, or the bispecific antibody of claim 135 or 136.

164. The method of claim 163, wherein the neurodegenerative disease is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Huntington's disease, Alzheimer's disease, primary age-related tauopathy, progressive supranuclear palsy (PSP), frontotemporal dementia, frontotemporal dementia with parkinsonism linked to chromosome 17, argyrophilic grain dementia, amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam, corticobasal degeneration, chronic traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia pugilistica, diffuse neurofibrillary tangles with calcification, Down's syndrome, familial British dementia, familial Danish dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean PSP, Hallevorden-Spatz disease, inclusion-body myositis, myotonic dystrophy, neurofibrillary tangle-predominant dementia, Niemann-Pick disease type C, pallido-ponto-nigral degeneration, Pick's disease, postencephalitic parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, subacute sclerosing panencephalitis, and tangle only dementia.

165. The method of claim 163 or 164, wherein the neurodegenerative disease is a synucleinopathy.

166. The method of claim 165, wherein the synucleinopathy is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy.

167. The method of any one of claims 163 to 166, wherein the subject is treated during an early stage of the neurodegenerative disease.

168. The method of any one of claims 163 to 167, wherein the subject has not been treated for the neurodegenerative disease, has not been treated with a dopaminergic replacement therapy, or has not been treated with a dopamine agonist therapy.

169. The method of any one of claims 163 to 168, wherein the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is administered to the subject is determined based on an amount of total human alpha-synuclein protein in a sample and/or an amount of human alpha-synuclein protein in a sample that is bound to the isolated antibody or bispecific antibody.

170. The method of claim 169, wherein the sample is obtained from the subject.

171. The method of claim 169 or 170, wherein the sample is selected from the group consisting of an interstitial brain fluid sample, a cerebrospinal fluid (CSF) sample, a blood sample, and a combination thereof.

172. The method of any one of claims 163 to 171, wherein the amount of the isolated antibody, the pharmaceutical composition, or the bispecific antibody that is administered to the subject is based on a level of neurodegenerative disease progression in the subject and/or an amount of human alpha-synuclein protein spreading in the brain of the subject.

173. The method of claim 172, wherein an imaging method is used to determine the level of neurodegenerative disease progression in the subject and/or the amount of human alpha-synuclein protein spreading in the brain of the subject.

174. The method of claim 173, wherein the imaging method is selected from the group consisting of positron emission tomography (PET) imaging, dopamine transporter (DaT) imaging, magnetic resonance imaging (MRI), and computed tomography (CT) imaging.

175. The method of claim 174, wherein PET imaging comprises using a PET probe that comprises a diabody of the antibody of any one of claims 1 to 133.