US2018371071A1PendingUtilityA1
Therapeutic DLL4 Binding Proteins
Est. expiryAug 29, 2029(~3.1 yrs left)· nominal 20-yr term from priority
Inventors:Lorenzo BenatuilErwin R. BoghaertJijie GuMaria HarrisJonathan A. HicksonChung-Ming HsiehYuliya KutskovaYingchun LiZhihong LiuSusan Morgan-Lappe
A61P 43/00A61P 9/00A61P 5/00A61P 9/10A61P 7/10A61P 9/14A61P 35/04A61P 27/02A61P 29/00A61P 35/00A61P 35/02A61P 25/00A61P 31/04A61P 13/08A61P 21/00A61P 11/04A61P 11/00A61P 1/16A61P 13/10A61P 13/12A61P 13/02A61P 17/06A61P 19/02A61P 1/18A61P 17/00A61P 17/02A61P 1/04A61P 15/00C07K 2317/567A61K 2039/505A61K 39/3955C07K 2317/92C07K 2317/73C07K 16/22C07K 7/06C07K 16/18C07K 2317/76C07K 2317/565C07K 2317/94A61K 45/06C07K 2317/21C07K 14/47C07K 2317/33C07K 16/2857C07K 7/08
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Claims
Abstract
Improved DLL4 binding proteins are described, including antibodies, CDR-grafted antibodies, human antibodies, and DLL4 binding fragments thereof, proteins that bind DLL4 with high affinity, and DLL4 binding proteins that neutralize DLL4 activity. The DLL4 binding proteins are useful for treating or preventing cancers and tumors and especially for treating or preventing tumor angiogenesis, and/or other angiogenesis-dependent diseases such as ocular neovascularization, or angiogenesis-independent diseases characterized by aberrant DLL4 expression or activity such as autoimmune disorders including multiple sclerosis.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . A binding protein comprising an antigen binding domain capable of binding human DLL4, said antigen binding domain comprising at least one or more CDRs selected from the group consisting of:
CDR-H1:
(SEQ ID NO: 99)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 ,
wherein;
X 1 is S or N;
X 2 is S, G, or N;
X 3 is S, N, T, G, or R;
X 4 is Y;
X 5 is Y or H;
X 6 is W; and
X 7 is G;
CDR-H2:
(SEQ ID NO: 100)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 -X 11 -X 12 -X 13 -X 14 -
X 15 -X 16 ,
wherein;
X 1 is D;
X 2 is I;
X 3 is Y, N, or S;
X 4 is Y;
X 5 is T, N, A, I, S, or R;
X 6 is G;
X 7 is S, N, T, or G;
X 8 is T;
X 9 is Y;
X 10 is Y;
X 11 is N;
X 12 is P;
X 13 is S;
X 14 is L;
X 15 is K; and
X 16 is S, N, D, or G;
CDR-H3:
(SEQ ID NO: 101)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 -X 11 ,
wherein;
X 1 is E, Y, F, Q, W, L, or A;
X 2 is D, A, S, G, V, E, or N;
X 3 is V, M, L, P, or A;
X 4 is I, A, P, R, S, K, Q, V, G, M, or E;
X 5 is L, Y, F, or M;
X 6 is R, G, S, Q, or A;
X 7 is G;
X 8 is G, A, or S;
X 9 is S, A, L, V, R, or G;
X 10 is D; and
X 11 is Y, D, S, N, H, E, R, L, P, C, I, M, T, Q, or K;
CDR-L1:
(SEQ ID NO: 102)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 -X 11 ,
wherein;
X 1 is S;
X 2 is G;
X 3 is Q, E, or D;
X 4 is R, S, G, M, K, L, or T;
X 5 is L;
X 6 is G;
X 7 is D or E;
X 8 is K;
X 9 is Y;
X 10 is A or V; and
X 11 is S;
CDR-L2:
(SEQ ID NO: 103)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 ,
wherein;
X 1 is E or Q;
X 2 is D;
X 3 is S, L, T, A, E, or F;
X 4 is K, T, E, N, Q, S, or M;
X 5 is R;
X 6 is P; and
X 7 is S;
and
CDR-L3:
(SEQ ID NO: 104)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 ,
wherein;
X 1 is Q;
X 2 is A;
X 3 is W;
X 4 is D;
X 5 is R, S, M, E, N, G, or K;
X 6 is D or E;
X 7 is T, V, A, S, or M;
X 8 is G, A, or C; and
X 9 is V.
2 . The binding protein according to claim 1 , wherein said at least one CDR comprises an amino acid sequence selected from the group consisting of:
residues 31-37 of SEQ ID NO:1 (CDR-H1); residues 52-67 of SEQ ID NO:1 (CDR-H2); residues 100-110 of SEQ ID NO:1 (CDR-H3); residues 23-33 of SEQ ID NO:111 (CDR-L1); residues 49-55 of SEQ ID NO:111 (CDR-L2); residues 88-96 of SEQ ID NO:111 (CDR-L3); SEQ ID NO:117 (CDR-H1); SEQ ID NO:118 (CDR-H2); SEQ ID NO:119 (CDR-H3); SEQ ID NO:121 (CDR-H1); SEQ ID NO:122 (CDR-H2); SEQ ID NO:123 (CDR-H3); SEQ ID NO:125 (CDR-H1); SEQ ID NO:126 (CDR-H2); SEQ ID NO:127 (CDR-H3); SEQ ID NO:129 (CDR-H1); SEQ ID NO:130 (CDR-H2); SEQ ID NO:131 (CDR-H3); SEQ ID NO:133 (CDR-H1); SEQ ID NO:134 (CDR-H2); SEQ ID NO:135 (CDR-H3); SEQ ID NO:137 (CDR-H1); SEQ ID NO:138 (CDR-H2); SEQ ID NO:139 (CDR-H3); SEQ ID NO:141 (CDR-H1); SEQ ID NO:142 (CDR-H2); SEQ ID NO:143 (CDR-H3); SEQ ID NO:145 (CDR-H1); SEQ ID NO:146 (CDR-H2); SEQ ID NO:147 (CDR-H3); SEQ ID NO:149 (CDR-H1); SEQ ID NO:150 (CDR-H2); SEQ ID NO:151 (CDR-H3); SEQ ID NO:153 (CDR-H1); SEQ ID NO:154 (CDR-H2); SEQ ID NO:155 (CDR-H3); SEQ ID NO:157 (CDR-H1); SEQ ID NO:158 (CDR-H2); SEQ ID NO:159 (CDR-H3); SEQ ID NO:161 (CDR-H1); SEQ ID NO:162 (CDR-H2); SEQ ID NO:163 (CDR-H3); SEQ ID NO:165 (CDR-H1); SEQ ID NO:166 (CDR-H2); SEQ ID NO:167 (CDR-H3); SEQ ID NO:169 (CDR-H1); SEQ ID NO:170 (CDR-H2); SEQ ID NO:171 (CDR-H3); SEQ ID NO:173 (CDR-H1); SEQ ID NO:174 (CDR-H2); SEQ ID NO:175 (CDR-H3); SEQ ID NO:177 (CDR-H1); SEQ ID NO:178 (CDR-H2); SEQ ID NO:179 (CDR-H3); SEQ ID NO:181 (CDR-H1); SEQ ID NO:182 (CDR-H2); SEQ ID NO:183 (CDR-H3); SEQ ID NO:185 (CDR-H1); SEQ ID NO:186 (CDR-H2); SEQ ID NO:187 (CDR-H3); SEQ ID NO:189 (CDR-H1); SEQ ID NO:190 (CDR-H2); SEQ ID NO:191 (CDR-H3); SEQ ID NO:193 (CDR-H1); SEQ ID NO:194 (CDR-H2); SEQ ID NO:195 (CDR-H3); SEQ ID NO:197 (CDR-H1); SEQ ID NO:198 (CDR-H2); SEQ ID NO:199 (CDR-H3); SEQ ID NO:201 (CDR-H1); SEQ ID NO:202 (CDR-H2); SEQ ID NO:203 (CDR-H3); SEQ ID NO:205 (CDR-H1); SEQ ID NO:206 (CDR-H2); SEQ ID NO:207 (CDR-H3); SEQ ID NO:209 (CDR-H1); SEQ ID NO:210 (CDR-H2); SEQ ID NO:211 (CDR-H3); SEQ ID NO:213 (CDR-H1); SEQ ID NO:214 (CDR-H2); SEQ ID NO:215 (CDR-H3); SEQ ID NO:217 (CDR-L1); SEQ ID NO:218 (CDR-L2); SEQ ID NO:219 (CDR-L3); SEQ ID NO:221 (CDR-L1); SEQ ID NO:222 (CDR-L2); SEQ ID NO:223 (CDR-L3); SEQ ID NO:225 (CDR-L1); SEQ ID NO:226 (CDR-L2); SEQ ID NO:227 (CDR-L3); SEQ ID NO:229 (CDR-L1); SEQ ID NO:230 (CDR-L2); SEQ ID NO:231 (CDR-L3); SEQ ID NO:233 (CDR-L1); SEQ ID NO:234 (CDR-L2); SEQ ID NO:235 (CDR-L3); SEQ ID NO:237 (CDR-L1); SEQ ID NO:238 (CDR-L2); SEQ ID NO:239 (CDR-L3); SEQ ID NO:241 (CDR-L1); SEQ ID NO:242 (CDR-L2); SEQ ID NO:243 (CDR-L3); SEQ ID NO:245 (CDR-L1); SEQ ID NO:246 (CDR-L2); SEQ ID NO:247 (CDR-L3); SEQ ID NO:249 (CDR-L1); SEQ ID NO:250 (CDR-L2); SEQ ID NO:251 (CDR-L3); SEQ ID NO:253 (CDR-L1); SEQ ID NO:254 (CDR-L2); SEQ ID NO:255 (CDR-L3); SEQ ID NO:257 (CDR-L1); SEQ ID NO:258 (CDR-L2); SEQ ID NO:259 (CDR-L3); SEQ ID NO:261 (CDR-L1); SEQ ID NO:262 (CDR-L2); SEQ ID NO:263 (CDR-L3); SEQ ID NO:265 (CDR-L1); SEQ ID NO:266 (CDR-L2); SEQ ID NO:267 (CDR-L3); SEQ ID NO:269 (CDR-L1); SEQ ID NO:270 (CDR-L2); SEQ ID NO:271 (CDR-L3); SEQ ID NO:273 (CDR-L1); SEQ ID NO:274 (CDR-L2); SEQ ID NO:275 (CDR-L3); SEQ ID NO:277 (CDR-L1); SEQ ID NO:278 (CDR-L2); SEQ ID NO:279 (CDR-L3); SEQ ID NO:281 (CDR-L1); SEQ ID NO:282 (CDR-L2); SEQ ID NO:283 (CDR-L3); SEQ ID NO:285 (CDR-L1); SEQ ID NO:286 (CDR-L2); SEQ ID NO:287 (CDR-L3); SEQ ID NO:289 (CDR-L1); SEQ ID NO:290 (CDR-L2); SEQ ID NO:291 (CDR-L3); SEQ ID NO:293 (CDR-L1); SEQ ID NO:294 (CDR-L2); SEQ ID NO:295 (CDR-L3); SEQ ID NO:297 (CDR-L1); SEQ ID NO:298 (CDR-L2); SEQ ID NO:299 (CDR-L3); SEQ ID NO:301 (CDR-L1); SEQ ID NO:302 (CDR-L2); SEQ ID NO:303 (CDR-L3); SEQ ID NO:305 (CDR-L1); SEQ ID NO:306 (CDR-L2); SEQ ID NO:307 (CDR-L3); SEQ ID NO:309 (CDR-L1); SEQ ID NO:310 (CDR-L2); SEQ ID NO:311 (CDR-L3); SEQ ID NO:313 (CDR-L1); SEQ ID NO:314 (CDR-L2); SEQ ID NO:315 (CDR-L3); residues 31-37 of SEQ ID NO:334 (CDR-H1); residues 52-67 of SEQ ID NO:334 (CDR-H2); residues 100-110 of SEQ ID NO:334 (CDR-H3); residues 23-33 of SEQ ID NO:335 (CDR-L1); residues 49-55 of SEQ ID NO:335 (CDR-L2); residues 88-96 of SEQ ID NO:335 (CDR-L3); residues 31-37 of SEQ ID NO:336 (CDR-H1); residues 52-67 of SEQ ID NO:336 (CDR-H2); residues 100-110 of SEQ ID NO:336 (CDR-H3); residues 23-33 of SEQ ID NO:337 (CDR-L1); residues 49-55 of SEQ ID NO:337 (CDR-L2); residues 88-96 of SEQ ID NO:337 (CDR-L3); residues 31-37 of SEQ ID NO:338 (CDR-H1); residues 52-67 of SEQ ID NO:338 (CDR-H2); residues 100-110 of SEQ ID NO:338 (CDR-H3); residues 23-33 of SEQ ID NO:339 (CDR-L1); residues 49-55 of SEQ ID NO:339 (CDR-L2); residues 88-96 of SEQ ID NO:339 (CDR-L3); residues 31-37 of SEQ ID NO:340 (CDR-H1); residues 52-67 of SEQ ID NO:340 (CDR-H2); residues 100-110 of SEQ ID NO:340 (CDR-H3); residues 23-33 of SEQ ID NO:341 (CDR-L1); residues 49-55 of SEQ ID NO:341 (CDR-L2); residues 88-96 of SEQ ID NO:341 (CDR-L3); residues 31-37 of SEQ ID NO:342 (CDR-H1); residues 52-67 of SEQ ID NO:342 (CDR-H2); residues 100-110 of SEQ ID NO:342 (CDR-H3); residues 23-33 of SEQ ID NO:343 (CDR-L1); residues 49-55 of SEQ ID NO:343 (CDR-L2); residues 88-96 of SEQ ID NO:343 (CDR-L3); residues 31-37 of SEQ ID NO:344 (CDR-H1); residues 52-67 of SEQ ID NO:344 (CDR-H2); residues 100-110 of SEQ ID NO:344 (CDR-H3); residues 24-34 of SEQ ID NO:345 (CDR-L1); residues 50-56 of SEQ ID NO:345 (CDR-L2); residues 89-97 of SEQ ID NO:345 (CDR-L3); residues 31-37 of SEQ ID NO:346 (CDR-H1); residues 52-67 of SEQ ID NO:346 (CDR-H2); residues 100-110 of SEQ ID NO:346 (CDR-H3); residues 23-33 of SEQ ID NO:347 (CDR-L1); residues 49-55 of SEQ ID NO:347 (CDR-L2); residues 88-96 of SEQ ID NO:347 (CDR-L3); residues 31-37 of SEQ ID NO:348 (CDR-H1); residues 52-67 of SEQ ID NO:348 (CDR-H2); residues 100-110 of SEQ ID NO:348 (CDR-H3); residues 24-34 of SEQ ID NO:349 (CDR-L1); residues 50-56 of SEQ ID NO:349 (CDR-L2); residues 89-97 of SEQ ID NO:349 (CDR-L3); residues 31-37 of SEQ ID NO:350 (CDR-H1); residues 52-67 of SEQ ID NO:350 (CDR-H2); residues 100-110 of SEQ ID NO:350 (CDR-H3); residues 24-34 of SEQ ID NO:351 (CDR-L1); residues 50-56 of SEQ ID NO:351 (CDR-L2); residues 89-97 of SEQ ID NO:351 (CDR-L3); residues 31-37 of SEQ ID NO:352 (CDR-H1); residues 52-67 of SEQ ID NO:352 (CDR-H2); residues 100-110 of SEQ ID NO:352 (CDR-H3); residues 24-34 of SEQ ID NO:353 (CDR-L1); residues 50-56 of SEQ ID NO:353 (CDR-L2); residues 89-97 of SEQ ID NO:353 (CDR-L3); residues 31-37 of SEQ ID NO:354 (CDR-H1); residues 52-67 of SEQ ID NO:354 (CDR-H2); residues 100-110 of SEQ ID NO:354 (CDR-H3); residues 24-34 of SEQ ID NO:355 (CDR-L1); residues 50-56 of SEQ ID NO:355 (CDR-L2); residues 89-97 of SEQ ID NO:355 (CDR-L3); residues 31-37 of SEQ ID NO:356 (CDR-H1); residues 52-67 of SEQ ID NO:356 (CDR-H2); residues 100-110 of SEQ ID NO:356 (CDR-H3); residues 23-33 of SEQ ID NO:357 (CDR-L1); residues 49-55 of SEQ ID NO:357 (CDR-L2); residues 88-96 of SEQ ID NO:357 (CDR-L3); residues 31-37 of SEQ ID NO:358 (CDR-H1); residues 52-67 of SEQ ID NO:358 (CDR-H2); residues 100-110 of SEQ ID NO:358 (CDR-H3); residues 23-33 of SEQ ID NO:359 (CDR-L1); residues 49-55 of SEQ ID NO:359 (CDR-L2); residues 88-96 of SEQ ID NO:359 (CDR-L3); residues 31-37 of SEQ ID NO:36((CDR-H1); residues 52-67 of SEQ ID NO:360 (CDR-H2); residues 100-110 of SEQ ID NO:360 (CDR-H3); residues 23-33 of SEQ ID NO:361 (CDR-L1); residues 49-55 of SEQ ID NO:361 (CDR-L2); residues 88-96 of SEQ ID NO:361 (CDR-L3).
3 . The binding protein according to claim 2 , wherein said binding protein comprises at least 3 CDRs.
4 . The binding protein according to claim 3 , wherein said at least 3 CDRs comprises a variable domain CDR set selected from the group consisting of:
VH E9 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:1 CDR-H2: residues 52-67 of SEQ ID NO:1 CDR-H3 residues 100-110 of SEQ ID NO:1 VL E9 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:111 CDR-L2: residues 49-55 of SEQ ID NO:111 CDR-L3: residues 88-96 of SEQ ID NO:111 VH E9.4 CDR Set CDR-H1: SEQ ID NO:117 CDR-H2: SEQ ID NO:118 CDR-H3: SEQ ID NO:119 VL E9.4 CDR Set CDR-L1: SEQ ID NO:229 CDR-L2: SEQ ID NO:230 CDR-L3: SEQ ID NO:231 VH E9.11 CDR Set CDR-H1: SEQ ID NO:121 CDR-H2: SEQ ID NO:122 CDR-H3: SEQ ID NO:123 VL E9.11 CDR Set CDR-L1: SEQ ID NO:233 CDR-L2: SEQ ID NO:234 CDR-L3: SEQ ID NO:235 VH E9.14 CDR Set CDR-H1: SEQ ID NO:125 CDR-H2: SEQ ID NO:126 CDR-H3: SEQ ID NO:127 VL E9.14 CDR Set CDR-L1: SEQ ID NO:237 CDR-L2: SEQ ID NO:238 CDR-L3: SEQ ID NO:239 VH E9.17 CDR Set CDR-H1: SEQ ID NO:129 CDR-H2: SEQ ID NO:130 CDR-H3: SEQ ID NO:131 VL E9.17 CDR Set CDR-L1: SEQ ID NO:241 CDR-L2: SEQ ID NO:242 CDR-L3: SEQ ID NO:243 VH E9.18 CDR Set CDR-H1: SEQ ID NO:133 CDR-H2: SEQ ID NO:134 CDR-H3: SEQ ID NO:135 VL E9.18 CDR Set CDR-L1: SEQ ID NO:245 CDR-L2: SEQ ID NO:246 CDR-L3: SEQ ID NO:247 VH E9.19 CDR Set CDR-H1: SEQ ID NO:137 CDR-H2: SEQ ID NO:138 CDR-H3: SEQ ID NO:139 VL E9.19 CDR Set CDR-L1: SEQ ID NO:249 CDR-L2: SEQ ID NO:250 CDR-L3: SEQ ID NO:251 VH E9.22 CDR Set CDR-H1: SEQ ID NO:141 CDR-H2: SEQ ID NO:142 CDR-H3: SEQ ID NO:143 VL E9.22 CDR Set CDR-L1: SEQ ID NO:253 CDR-L2: SEQ ID NO:254 CDR-L3: SEQ ID NO:255 VH E9.48 CDR Set CDR-H1: SEQ ID NO:145 CDR-H2: SEQ ID NO:146 CDR-H3: SEQ ID NO:147 VL E9.48 CDR Set CDR-L1: SEQ ID NO:257 CDR-L2: SEQ ID NO:258 CDR-L3: SEQ ID NO:259 VH E9.65 CDR Set CDR-H1: SEQ ID NO:149 CDR-H2: SEQ ID NO:150 CDR-H3: SEQ ID NO:151 VL E9.65 CDR Set CDR-L1: SEQ ID NO:261 CDR-L2: SEQ ID NO:262 CDR-L3: SEQ ID NO:263 VH E9.66 CDR Set CDR-H1: SEQ ID NO:153 CDR-H2: SEQ ID NO:154 CDR-H3: SEQ ID NO:155 VL E9.66 CDR Set CDR-L1: SEQ ID NO:265 CDR-L2: SEQ ID NO:266 CDR-L3: SEQ ID NO:267 VH E9.71 CDR Set CDR-H1: SEQ ID NO:157 CDR-H2: SEQ ID NO:158 CDR-H3: SEQ ID NO:159 VL E9.71 CDR Set CDR-L1: SEQ ID NO:269 CDR-L2: SEQ ID NO:270 CDR-L3: SEQ ID NO:271 VH E9.13 CDR Set CDR-H1: SEQ ID NO:161 CDR-H2: SEQ ID NO:162 CDR-H3: SEQ ID NO:163 VL E9.13 CDR Set CDR-L1: SEQ ID NO:217 CDR-L2: SEQ ID NO:218 CDR-L3: SEQ ID NO:219 VH E9.16 CDR Set CDR-H1: SEQ ID NO:165 CDR-H2: SEQ ID NO:166 CDR-H3: SEQ ID NO:167 VL E9.16 CDR Set CDR-L1: SEQ ID NO:221 CDR-L2: SEQ ID NO:222 CDR-L3: SEQ ID NO:223 VH E9.38 CDR Set CDR-H1: SEQ ID NO:169 CDR-H2: SEQ ID NO:170 CDR-H3: SEQ ID NO:171 VL E9.38 CDR Set CDR-L1: SEQ ID NO:225 CDR-L2: SEQ ID NO:226 CDR-L3: SEQ ID NO:227 VH E9.2B CDR Set CDR-H1: SEQ ID NO:173 CDR-H2: SEQ ID NO:174 CDR-H3: SEQ ID NO:175 VL E9.2B CDR Set CDR-L1: SEQ ID NO:273 CDR-L2: SEQ ID NO:274 CDR-L3: SEQ ID NO:275 VH E9.1F CDR Set CDR-H1: SEQ ID NO:177 CDR-H2: SEQ ID NO:178 CDR-H3: SEQ ID NO:179 VL E9.1F CDR Set CDR-L1: SEQ ID NO:277 CDR-L2: SEQ ID NO:278 CDR-L3: SEQ ID NO:279 VH E9.10H CDR Set CDR-H1: SEQ ID NO:181 CDR-H2: SEQ ID NO:182 CDR-H3: SEQ ID NO:183 VL E9.10H CDR Set CDR-L1: SEQ ID NO:301 CDR-L2: SEQ ID NO:302 CDR-L3: SEQ ID NO:303 VH E9.5E CDR Set CDR-H1: SEQ ID NO:185 CDR-H2: SEQ ID NO:186 CDR-H3: SEQ ID NO:187 VL E9.5E CDR Set CDR-L1: SEQ ID NO:293 CDR-L2: SEQ ID NO:294 CDR-L3: SEQ ID NO:295 VH E9.10C CDR Set CDR-H1: SEQ ID NO:189 CDR-H2: SEQ ID NO:190 CDR-H3: SEQ ID NO:191 VL E9.10C CDR Set CDR-L1: SEQ ID NO:281 CDR-L2: SEQ ID NO:282 CDR-L3: SEQ ID NO:283 VH E9.7E CDR Set CDR-H1: SEQ ID NO:193 CDR-H2: SEQ ID NO:194 CDR-H3: SEQ ID NO:195 VL E9.7E CDR Set CDR-L1: SEQ ID NO:289 CDR-L2: SEQ ID NO:290 CDR-L3: SEQ ID NO:291 VH E9.12B CDR Set CDR-H1: SEQ ID NO:197 CDR-H2: SEQ ID NO:198 CDR-H3: SEQ ID NO:199 VL E9.12B CDR Set CDR-L1: SEQ ID NO:297 CDR-L2: SEQ ID NO:298 CDR-L3: SEQ ID NO:299 VH E9.10E CDR Set CDR-H1: SEQ ID NO:201 CDR-H2: SEQ ID NO:202 CDR-H3: SEQ ID NO:203 VL E9.10E CDR Set CDR-L1: SEQ ID NO:285 CDR-L2: SEQ ID NO:286 CDR-L3: SEQ ID NO:287 VH E9.6A CDR Set CDR-H1: SEQ ID NO:205 CDR-H2: SEQ ID NO:206 CDR-H3: SEQ ID NO:207 VL E9.6A CDR Set CDR-L1: SEQ ID NO:305 CDR-L2: SEQ ID NO:306 CDR-L3: SEQ ID NO:307 VH E9.7A CDR Set CDR-H1: SEQ ID NO:209 CDR-H2: SEQ ID NO:210 CDR-H3: SEQ ID NO:211 VL E9.7A CDR Set CDR-L1: SEQ ID NO:309 CDR-L2: SEQ ID NO:310 CDR-L3: SEQ ID NO:311 VH E9.8H CDR Set CDR-H1: SEQ ID NO:213 CDR-H2: SEQ ID NO:214 CDR-H3: SEQ ID NO:215 VL E9.8H CDR Set CDR-L1: SEQ ID NO:313 CDR-L2: SEQ ID NO:314 CDR-L3: SEQ ID NO:315 VH E9.1 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:334 CDR-H2: residues 52-67 of SEQ ID NO:334 CDR-H3: residues 100-110 of SEQ ID NO:334 VL E9.1 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:335 CDR-L2: residues 49-55 of SEQ ID NO:335 CDR-L3: residues 88-96 of SEQ ID NO:335 VH E9-SE1 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:336 CDR-H2: residues 52-67 of SEQ ID NO:336 CDR-H3: residues 100-110 of SEQ ID NO:336 VL E9-SE CDR Set CDR-L1: residues 23-33 of SEQ ID NO:337 CDR-L2: residues 49-55 of SEQ ID NO:337 CDR-L3: residues 88-96 of SEQ ID NO:337 VH E9-SE2 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:338 CDR-H2: residues 52-67 of SEQ ID NO:338 CDR-H3: residues 100-110 of SEQ ID NO:338 VL E9-SE2 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:339 CDR-L2: residues 49-55 of SEQ ID NO:339 CDR-L3: residues 88-96 of SEQ ID NO:339 VH E9-SE3 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:340 CDR-H2: residues 52-67 of SEQ ID NO:340 CDR-H3: residues 100-110 of SEQ ID NO:340 VL E9-SE3 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:341 CDR-L2: residues 49-55 of SEQ ID NO:341 CDR-L3: residues 88-96 of SEQ ID NO:341 VH E9-SE4 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:342 CDR-H2: residues 52-67 of SEQ ID NO:342 CDR-H3: residues 100-110 of SEQ ID NO:342 VL E9-SE4 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:343 CDR-L2: residues 49-55 of SEQ ID NO:343 CDR-L3: residues 88-96 of SEQ ID NO:343 VH E9-SE5 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:344 CDR-H2: residues 52-67 of SEQ ID NO:344 CDR-H3: residues 100-110 of SEQ ID NO:344 VL E9-SE5 CDR Set CDR-L1: residues 24-34 of SEQ ID NO:345 CDR-L2: residues 50-56 of SEQ ID NO:345 CDR-L3: residues 89-97 of SEQ ID NO:345 VH E9-SE6 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:346 CDR-H2: residues 52-67 of SEQ ID NO:346 CDR-H3: residues 100-110 of SEQ ID NO:346 VL E9-SE6 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:347 CDR-L2: residues 49-55 of SEQ ID NO:347 CDR-L3: residues 88-96 of SEQ ID NO:347 VH E9-SE7 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:348 CDR-H2: residues 52-67 of SEQ ID NO:348 CDR-H3: residues 100-110 of SEQ ID NO:348 VL E9-SE7 CDR Set CDR-L1: residues 24-34 of SEQ ID NO:349 CDR-L2: residues 50-56 of SEQ ID NO:349 CDR-L3: residues 89-97 of SEQ ID NO:349 VH E9-SE8 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:350 CDR-H2: residues 52-67 of SEQ ID NO:350 CDR-H3: residues 100-110 of SEQ ID NO:350 VL E9-SE8 CDR Set CDR-L1: residues 24-34 of SEQ ID NO:351 CDR-L2: residues 50-56 of SEQ ID NO:351 CDR-L3: residues 89-97 of SEQ ID NO:351 VH E9-FR1 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:352 CDR-H2: residues 52-67 of SEQ ID NO:352 CDR-H3: residues 100-110 of SEQ ID NO:352 VL E9-FR1 CDR Set CDR-L1: residues 24-34 of SEQ ID NO:353 CDR-L2: residues 50-56 of SEQ ID NO:353 CDR-L3: residues 89-97 of SEQ ID NO:353 VH E9-FR2 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:354 CDR-H2: residues 52-67 of SEQ ID NO:354 CDR-H3: residues 100-110 of SEQ ID NO:354 VL E9-FR2 CDR Set CDR-L1: residues 24-34 of SEQ ID NO:355 CDR-L2: residues 50-56 of SEQ ID NO:355 CDR-L3: residues 89-97 of SEQ ID NO:355 VH E9.71 CDR Set CDR-H1: residues 31-37 of SEQ ID NO:356 CDR-H2: residues 52-67 of SEQ ID NO:356 CDR-H3: residues 100-110 of SEQ ID NO:356 VL E9.71 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:357 CDR-L2: residues 49-55 of SEQ ID NO:357 CDR-L3: residues 88-96 of SEQ ID NO:357 VH E9.71(M) CDR Set CDR-H1: residues 31-37 of SEQ ID NO:358 CDR-H2: residues 52-67 of SEQ ID NO:358 CDR-H3: residues 100-110 of SEQ ID NO:358 VL E9.71(M) CDR Set CDR-L1: residues 23-33 of SEQ ID NO:359 CDR-L2: residues 49-55 of SEQ ID NO:359 CDR-L3: residues 88-96 of SEQ ID NO:359 VH E9.71(L) CDR Set CDR-H1: residues 31-37 of SEQ ID NO:360 CDR-H2: residues 52-67 of SEQ ID NO:360 CDR-H3: residues 100-110 of SEQ ID NO:360 VL E9.71(L) CDR Set CDR-L1: residues 23-33 of SEQ ID NO:361 CDR-L2: residues 49-55 of SEQ ID NO:361 CDR-L3: residues 88-96 of SEQ ID NO:361
5 . The binding protein according to claim 4 , comprising at least two variable domain CDR sets.
6 . The binding protein according to claim 5 , wherein said at least two variable domain CDR sets are selected from a group consisting of:
VH E9 CDR Set and VL E9 CDR Set, VH E9.4 CDR Set and VL E9.4 CDR Set, VH E9.11 CDR Set and VL E9.11 CDR Set, VH E9.14 CDR Set and VL E9.14 CDR Set, VH E9.17 CDR Set and VL E9.17 CDR Set, VH E9.18 CDR Set and VL E9.18 CDR Set, VH E9.19 CDR Set and VL E9.19 CDR Set, VH E9.22 CDR Set and VL E9.22 CDR Set, VH E9.48 CDR Set and VL E9.48 CDR Set, VH E9.65 CDR Set and VL E9.65 CDR Set, VH E9.66 CDR Set and VL E9.66 CDR Set, VH E9.71 CDR Set and VL E9.71 CDR Set, VH E9.13 CDR Set and VL E9.13 CDR Set, VH E9.16 CDR Set and VL E9.16 CDR Set, VH E9.38 CDR Set and VL E9.38 CDR Set, VH E9.2B CDR Set and VL E9.2B CDR Set, VH E9.1F CDR Set and VL E9.1F CDR Set, VH E9.10H CDR Set and VL E9.10H CDR Set, VH E9.5E CDR Set and VL E9.5E CDR Set, VH E9.10C CDR Set and VL E9.10C CDR Set, VH E9.7E CDR Set and VL E9.7E CDR Set, VH E9.12B CDR Set and VL E9.12B CDR Set, VH E9.10E CDR Set and VL E9.10E CDR Set, VH E9.6A CDR Set and VL E9.6A CDR Set, VH E9.7A CDR Set and VL E9.7A CDR Set, VH E9.8H CDR Set and VL E9.8H CDR Set, VH E9-SE1 CDR Set and VL E9-SE1 CDR Set, VH E9-SE2 CDR Set and VL E9-SE2 CDR Set, VH E9-SE3 CDR Set and VL E9-SE3 CDR Set, VH E9-SE4 CDR Set and VL E9-SE4 CDR Set, VH E9-SE5 CDR Set and VL E9-SE5 CDR Set, VH E9-SE6 CDR Set and VL E9-SE6 CDR Set, VH E9-SE7 CDR Set and VL E9-SE7 CDR Set, VH E9-SE8 CDR Set and VL E9-SE8 CDR Set, VH E9-FR1 CDR Set and VL E9-FR1 CDR Set, VH E9-FR2 CDR Set and VL E9-FR2 CDR Set, VH E9.71 CDR Set and VL E9.71 CDR Set, VH E9.71(M) CDR Set and VL E9.71(M) CDR Set, and VH E9.71(L) CDR Set and VL E9.71(L) CDR Set.
7 . The binding protein according to any one of claims 1 - 6 , further comprising a human acceptor framework.
8 . The binding protein according to claim 7 , wherein said human acceptor framework comprises an amino acid sequence selected from the group consisting of:
heavy chain acceptor framework sequences SEQ ID NOS:6-22, heavy chain acceptor sequences SEQ ID NOS:35-62, light chain acceptor sequences SEQ ID NOS:23-34, and light chain acceptor sequences SEQ ID NOS:63-98.
9 . The binding protein according to claim 7 or claim 8 , wherein said human acceptor framework comprises at least one framework region amino acid substitution, wherein the amino acid sequence of the framework is at least 65% identical to the sequence of said human acceptor framework and comprising at least 70 amino acid residues identical to said human acceptor framework.
10 . The binding protein according to claim 8 , wherein said human acceptor framework comprises at least one framework region amino acid substitution at a key residue, said key residue selected from the group consisting of:
a residue adjacent to a CDR; a glycosylation site residue; a rare residue; a residue capable of interacting with human DLL4 a canonical residue; a contact residue between heavy chain variable region and light chain variable region; a residue within a Vernier zone; and a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework.
11 . The binding protein according to claim 10 , wherein key residue selected from the group consisting of: 2H, 4H, 24H, 26H, 27H, 29H, 34H, 35H, 37H, 39H, 44H, 45H, 47H, 48H, 49H, 50H, 51H, 58H, 59H, 60H, 63H, 67H, 69H, 71H, 73H, 76H, 78H, 91H, 93H, 94H, 2L, 4L, 25L, 29L, 27bL, 33L, 34L, 36L, 38L, 43L, 44L, 46L, 47L, 48L, 49L, 55L, 58L, 62L, 64L, 71L, 87L, 89L, 90L, 91L, 94L, 95L.
12 . The binding protein according to claim 11 , where the binding protein is a consensus human variable domain.
13 . The binding protein according to claim 1 , where said binding protein comprises at least one variable domain having amino acid sequence selected from the group consisting of: SEQ ID NOS:1, 111, 116, 228, 120, 232, 124, 236, 128, 240, 132, 244, 136, 248, 140, 252, 144, 256, 148, 260, 152, 264, 156, 268, 160, 216, 164, 220, 168, 224, 172, 272, 176, 276, 180, 300, 184, 292, 188, 280, 192, 288, 196, 296, 200, 284, 204, 304, 208, 308, 212, 312, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, and 361.
14 . The binding protein according to claim 13 , wherein said binding protein comprises two variable domains, wherein said two variable domains have amino acid sequences selected from the group consisting of: SEQ ID NOS:1 and 111, SEQ ID NOS:116 and 228, SEQ ID NOS:120 and 232, SEQ ID NOS:124 and 236, SEQ ID NOS:128 and 240, SEQ ID NOS:132 and 244, SEQ ID NOS:136 and 248, SEQ ID NOS:140 and 252, SEQ ID NOS:144 and 256, SEQ ID NOS:148 and 260, SEQ ID NOS:152 and 264, SEQ ID NOS:156 and 268, SEQ ID NOS:160 and 216, SEQ ID NOS:164 and 220, SEQ ID NOS:168 and 224, SEQ ID NOS:172 and 272, SEQ ID NOS:176 and 276, SEQ ID NOS:180 and 300, SEQ ID NOS:184 and 292, SEQ ID NOS:188 and 280, SEQ ID NOS:192 and 288, SEQ ID NOS:196 and 296, SEQ ID NOS:200 and 284, SEQ ID NOS:204 and 304, SEQ ID NOS:208 and 308, SEQ ID NOS:212 and 312, SEQ ID NOS:334 and 335, SEQ ID NOS:336 and 337, SEQ ID NOS:338 and 339, SEQ ID NOS:340 and 341, SEQ ID NOS:342 and 343, SEQ ID NOS:344 and 345, SEQ ID NOS:346 and 347, SEQ ID NOS:348 and 349, SEQ ID NOS:350 and 351, SEQ ID NOS:352 and 353, SEQ ID NOS:354 and 355, SEQ ID NOS:356 and 357, SEQ ID NOS:358 and 359, SEQ ID NOS:360 and 361.
15 . The binding protein according to claim 11 , wherein said binding protein comprises at least one variable domain having an amino acid sequence selected from the group consisting of: SEQ ID NOS:1, 111, 116, 228, 120, 232, 124, 236, 128, 240, 132, 244, 136, 248, 140, 252, 144, 256, 148, 260, 152, 264, 156, 268, 160, 216, 164, 220, 168, 224, 172, 272, 176, 276, 180, 300, 184, 292, 188, 280, 192, 288, 196, 296, 200, 284, 204, 304, 208, 308, 212, 312, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, and 361.
16 . The binding protein according to claim 2 wherein said antigen binding domain comprises a V H .
17 . The binding protein according to claim 16 wherein said V H comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:1, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, 156, 160, 164, 168, 172, 176, 180, 184, 188, 192, 196, 200, 204, 208, 212, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, and 360.
18 . The binding protein according to claim 2 wherein said antigen binding domain comprises a V L .
19 . The binding protein according to claim 18 wherein said VL comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:111, 228, 232, 236, 240, 244, 248, 252, 256, 260, 264, 268, 216, 220, 224, 272, 276, 300, 292, 280, 288, 296, 284, 304, 308, 312, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, and 361.
20 . The binding protein according to claim 2 wherein said antigen binding domain comprises a V H and a V L .
21 . The binding protein according to claim 19 further comprising a V H wherein said V H comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:1, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, 156, 160, 164, 168, 172, 176, 180, 184, 188, 192, 196, 200, 204, 208, 212, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, and 360.
22 . The binding protein according to claim 20 wherein said VL comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:111, 228, 232, 236, 240, 244, 248, 252, 256, 260, 264, 268, 216, 220, 224, 272, 276, 300, 292, 280, 288, 296, 284, 304, 308, 312, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, and 361.
23 . The binding protein according to claim 2 , further comprising a heavy chain immunoglobulin constant domain selected from the group consisting of: a human IgM constant domain; a human IgG1 constant domain; a human IgG2 constant domain; a human IgG3 constant domain; a human IgG4 constant domain; a human IgE constant domain and a human IgA constant domain.
24 . The binding protein according to claim 23 wherein said heavy chain immunoglobulin constant region domain is a human IgG1 constant domain.
25 . The binding protein according to claim 24 wherein said human IgG1 constant domain comprises amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:3.
26 . The binding protein according to claim 2 , further comprising a light chain immunoglobulin constant domain selected from the group consisting of: a human Ig kappa constant domain and a human Ig lambda constant domain.
27 . The binding protein according to claim 26 wherein said light chain immunoglobulin constant region domain is a human Ig kappa constant domain comprising amino acid sequence SEQ ID NO:4.
28 . The binding protein according to claim 26 wherein said light chain immunoglobulin constant region domain is a human Ig lambda constant domain comprising amino acid sequence SEQ ID NO:5.
29 . The binding protein according to claim 1 wherein said binding protein is selected from the group consisting of: an immunoglobulin molecule, an scFv, a monoclonal antibody, a human antibody, a chimeric antibody, a humanized antibody, a single domain antibody, a Fab fragment, a Fab′ fragment, an F(ab′) 2 , an Fv, a disulfide linked Fv, a single domain antibody, a diabody, a multispecific antibody, a bispecific antibody, and a dual specific antibody.
30 . The binding protein according to claim 29 wherein said binding protein is a human antibody.
31 . A binding protein capable of binding human DLL-4, said binding protein comprising:
an Ig constant heavy region having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:3; an Ig constant light region having an amino acid sequence selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:5; an Ig variable heavy region having an amino acid sequence selected from the group consisting: SEQ ID NOS:1, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, 156, 160, 164, 168, 172, 176, 180, 184, 188, 192, 196, 200, 204, 208, 212, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, and 360; and an Ig variable light region having an amino acid sequence selected from the group consisting: SEQ ID NOS:111, 228, 232, 236, 240, 244, 248, 252, 256, 260, 264, 268, 216, 220, 224, 272, 276, 300, 292, 280, 288, 296, 284, 304, 308, 312, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, and 361.
32 . The binding protein according to claim 31 , wherein the Ig constant light region is SEQ ID NO:5.
33 . The binding protein according to any one of claims 1 - 32 , wherein the binding protein is capable of blocking DLL4 interaction with a Notch protein selected from the group consisting of Notch-1, Notch-2, Notch-3, Notch-4, and combinations thereof.
34 . The binding protein according to claim 33 , wherein the binding protein is capable of blocking DLL4 interaction with Notch-1 and Notch-4.
35 . The binding protein according to claim 33 , wherein the binding protein is capable of blocking DLL4 interaction with Notch-1.
36 . The binding protein according to claim 33 , wherein the binding protein is capable of blocking DLL4 interaction with Notch-4.
37 . The binding protein according to any one of claims 1 - 32 , wherein the binding protein is capable of modulating a biological function of DLL4.
38 . The binding protein according to any one of claims 1 - 32 , wherein said binding protein is capable of neutralizing a DLL4.
39 . The neutralizing binding protein according to claim 38 wherein said DLL4 is selected from the group consisting of: human DLL4, mouse DLL4, cynomolgus DLL4, and rat DLL4.
40 . The neutralizing binding protein according to claim 38 wherein said neutralizing binding protein diminishes the ability of DLL4 to bind to its receptor.
41 . The neutralizing binding protein according to claim 38 wherein said neutralizing binding protein is capable of reducing normal angiogenesis.
42 . The neutralizing binding protein according to claim 38 , wherein said neutralizing binding protein has a dissociation constant (K D ) selected from the group consisting of: at most about 10 −7 M; at most about 10 −8 M; at most about 10 −9 M; at most about 10 −10 M; at most about 10 −11 M; at most about 10 −12 M; and at most 10 −13 M.
43 . The neutralizing binding protein according to claim 38 , wherein said neutralizing binding protein has an on rate selected from the group consisting of: at least about 10 2 M −1 s −1 ; at least about 10 3 M −1 s −1 ; at least about 10 4 M −1 s −1 ; at least about 10 5 M −1 s −1 ; and at least about 10 6 M −1 s −1 .
44 . The neutralizing binding protein according to claim 38 , wherein said neutralizing binding protein has an off rate selected from the group consisting of: at most about 10 −3 s −1 ; at most about 10 −4 s −1 ; at most about 10 −5 s −1 ; and at most about 10 −6 s −1 .
45 . A labeled binding protein comprising a binding protein of any one of claims 1 - 32 , wherein said binding protein is conjugated to a detectable label.
46 . The labeled binding protein of claim 45 , wherein the detectable label is selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
47 . The labeled binding protein of claim 46 , wherein said label is a radiolabel selected from the group consisting of: 3 H, 14 C, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 137 Lu, 166 Ho, and 153 Sm.
48 . An antibody construct comprising a binding protein described in any of claims 1 - 32 and further comprising a linker polypeptide or an immunoglobulin constant domain.
49 . The antibody construct according to claim 48 , selected from the group consisting of:
an immunoglobulin molecules, a monoclonal antibody, a chimeric antibody, a CDR-grafted antibody, a humanized antibody, a Fab, a Fab′, a F(ab′) 2 , a Fv, a disulfide linked Fv, a scFv, a single domain antibody, a diabody, a multispecific antibody, a dual specific antibody, and a bispecific antibody.
50 . The antibody construct according to claim 48 , wherein said antibody construct comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:
a human IgM constant domain, a human IgG1 constant domain, a human IgG2 constant domain, a human IgG3 constant domain, a human IgG4 constant domain, a human IgE constant domain, a human IgA constant domain, and a IgG constant domain variant with one or more mutations altering binding strength to Fc neonatal receptor, Fc gamma receptors, or C1q.
51 . The antibody construct according to claim 48 , comprising an immunoglobulin constant domain having an amino acid sequence selected from consisting of: SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and combinations thereof.
52 . An antibody conjugate comprising an antibody construct as described in claim 48 , wherein said antibody construct is conjugated to a therapeutic or cytotoxic agent.
53 . The antibody conjugate of claim 52 , wherein said therapeutic or cytotoxic agent is selected from the group consisting of: an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.
54 . An isolated nucleic acid encoding a polypeptide selected from the group consisting of: a polypeptide comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises one or more of CDR-H1, a CDR-H2, or a CDR-H3 as described in claim 1 ; a polypeptide comprising a light chain variable domain, wherein the light chain variable domain comprises one or more of CDR-L1, a CDR-L2, or a CDR-L3 as described in claim 1 ; or a combination of both polypeptides.
55 . A vector comprising an isolated nucleic acid according to claim 54 .
56 . The vector of claim 55 wherein said vector is selected from the group consisting of: pcDNA, pTT, pTT3, pEFBOS, pBV, pJV, pHybE and pBJ.
57 . A host cell comprising a vector according to any one of claims 55 and 56 .
58 . The host cell according to claim 57 , wherein said host cell is a prokaryotic cell.
59 . The host cell according to claim 58 , wherein said host cell is Escherichia coli.
60 . The host cell according to claim 59 , wherein said host cell is a eukaryotic cell.
61 . The host cell according to claim 60 , wherein said eukaryotic cell is selected from the group consisting of: a protist cell, an animal cell, a plant cell, and a fungal cell.
62 . The host cell according to claim 61 , wherein said eukaryotic cell is an animal cell selected from the group consisting of: a mammalian cell, an avian cell, and an insect cell.
63 . The host cell according to claim 62 , wherein said mammalian cell is a CHO cell.
64 . The host cell according to claim 62 , wherein said mammalian cell is a COS cell.
65 . The host cell according to claim 61 , wherein said fungal cell is Saccharomyces cerevisiae.
66 . The host cell according to claim 62 , wherein said insect cell is an Sf9 cell.
67 . A method of producing a binding protein that binds human DLL4, comprising culturing the host cell of any one of claims 57 - 66 in a culture medium under conditions sufficient to produce a binding protein that binds human DLL4.
68 . A binding protein produced according to the method of claim 67 .
69 . A crystallized binding protein comprising a binding protein according to any one of claims 1 - 32 , wherein said binding protein exists as a crystal.
70 . The crystallized binding protein according to claim 69 , wherein said crystal is a carrier-free pharmaceutical controlled release crystal.
71 . The crystallized binding protein according to claim 69 , wherein said binding protein has a greater half life in vivo than the soluble counterpart of said binding protein.
72 . The crystallized binding protein according to claim 69 , wherein said binding protein retains biological activity.
73 . A composition for the release of a binding protein said composition comprising:
(a) a formulation, wherein said formulation comprises a crystallized binding protein, according to any one of claims 69 - 72 , and an ingredient; and (b) at least one polymeric carrier.
74 . The composition according to claim 73 , wherein said polymeric carrier is a polymer selected from one or more of the group consisting of: poly (acrylic acid), poly (cyanoacrylates), poly (amino acids), poly (anhydrides), poly (depsipeptide), poly (esters), poly (lactic acid), poly (lactic-co-glycolic acid) or PLGA, poly (b-hydroxybutryate), poly (caprolactone), poly (dioxanone), poly (ethylene glycol), poly ((hydroxypropyl) methacrylamide, poly [(organo)phosphazene], poly (ortho esters), poly (vinyl alcohol), poly (vinylpyrrolidone), maleic anhydride-alkyl vinyl ether copolymers, pluronic polyols, albumin, alginate, cellulose and cellulose derivatives, collagen, fibrin, gelatin, hyaluronic acid, oligosaccharides, glycaminoglycans, sulfated polyeaccharides, blends and copolymers thereof.
75 . The composition according to claim 73 , wherein said ingredient is selected from the group consisting of albumin, sucrose, trehalose, lactitol, gelatin, hydroxypropyl-β-cyclodextrin, methoxypolyethylene glycol and polyethylene glycol.
76 . A method for treating a mammal comprising the step of administering to the mammal an effective amount of the composition according to claim 73 .
77 . A pharmaceutical composition comprising the binding protein of any one of claims 1 - 32 , and a pharmaceutically acceptable carrier.
78 . The pharmaceutical composition of claim 77 which further comprises at least one additional therapeutic agent for treating a disorder in which DLL4 activity is detrimental.
79 . The pharmaceutical composition of claim 78 , wherein said additional agent is selected from the group consisting of: angiogenesis inhibitors; kinase inhibitors; co-stimulation molecule blockers; adhesion molecule blockers; anti-cytokine antibody or functional fragment thereof; methotrexate; corticosteroids; cyclosporine; rapamycin; FK506; and non-steroidal anti-inflammatory agents.
80 . A method for reducing human DLL4 activity comprising contacting human DLL4 with the binding protein of any one of claims 1 - 32 such that human DLL4 activity is reduced.
81 . A method for reducing human DLL4 activity in a human subject suffering from a disorder in which DLL4 activity is detrimental, comprising administering to the human subject the binding protein of any one of claims 1 - 32 such that human IL-17 activity in the human subject is reduced.
82 . A method for treating a subject for a disease or a disorder in which DLL4 activity is detrimental by administering to the subject the binding protein of any one of claims 1 - 32 such that treatment is achieved.
83 . The method of claim 82 , wherein said disorder is selected from the group consisting of: breast cancer, colon cancer, rectal cancer, lung cancer, oropharynx cancer, hypopharynx cancer, esophageal cancer, stomach cancer, pancreas cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, female genital tract cancer, male genital tract cancer, endocrine gland cancer, skin cancer, hemangiomas, melanomas, sarcomas, brain tumor, nerve cancer, eye tumor, meninges cancer, solid tumors from hematopoietic malignancies, tumor metastases, ocular neovascularization, edema, rheumatoid arthritis, multiple sclerosis, atheroscleorotic plaques, Crohn's disease, inflammatory bowel disease, refractory ascites, psoriasis, sarcoidosis, arterial arteriosclerosis, sepsis, peptic ulcers, burns, and pancreatitis, polycystic ovarian disease (POD), endometriosis, uterine fibroids, benign prostate hypertrophy, and other angiogenesis independent and dependent diseases characterized by abberant DLL4 activity.
84 . The method according to claim 83 , wherein the disorder is a primary and metastatic cancer.
85 . The method according to claim 83 , wherein the urinary tract cancer is selected from the group consisting of renal cancer, bladder cancer, and urothelium cancer.
86 . The method according to claim 83 , wherein the female genital tract cancer is selected from the group consisting of cervical cancer, uterine cancer, ovarian cancer, choriocarcinoma, and gestational trophoblastic disease.
87 . The method according to claim 83 , wherein the male genital tract cancer is selected from the group consisting of prostate cancer, seminal vesicles cancer, testicular cancer, and germ cell tumor.
88 . The method according to claim 83 , wherein the endocrine gland cancer is selected from the group consisting of thyroid cancer, adrenal cancer, and pituitary gland cancer.
89 . The method according to claim 83 , wherein the sarcoma is selected from the group consisting of a bone sarcoma, a soft tissue sarcoma, and Kaposi's sarcoma.
90 . The method according to claim 83 , wherein the meninges cancer is selected from the group consisting of an astrocytoma, a glioma, a glioblastoma, a retinoblastoma, a neuroma, a neuroblastoma, a Schwannoma, and a meningiomas.
91 . The method according to claim 83 , wherein the solid tumor from a hematopoietic malignancy is a leukemia, a Hodgkin's leukemia, a non-Hodgkin's leukemia, a lymphoma, a Hodgkin's lymphoma, and a non-Hodgkin's lymphomas.
92 . The method according to claim 83 , wherein the ocular neovascularization is selected from the group consisting of diabetic blindness, a retinopathy, an age-induced macular degeneration, and a rubeosis.
93 . A method of treating a patient suffering from a disorder in which DLL4 is detrimental comprising the step of administering the binding protein of any one of claims 1 - 32 before, concurrent with, or after the administration of a second agent, wherein the second agent is selected from the group consisting of an antibody or fragment thereof capable of binding human VEGFR2; methotrexate; an antibody, or fragment thereof, capable of binding human TNF; corticosteroids, cyclosporine, rapamycin, FK506, and non-steroidal anti-inflammatory agents.
94 . A binding protein comprising an antigen binding domain capable of binding human DLL4, said antigen binding domain comprising at least one or more CDRs selected from the group consisting of:
CDR-H1:
(SEQ ID NO: 105)
X 1 -X 2 -X 3 -X 4 -X 5 ,
wherein;
X 1 is S, N, or D;
X 2 is H or Y;
X 3 is W;
X 4 is M;
X 5 is S or H;
CDR-H2:
(SEQ ID NO: 106)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 -X 11 -X 12 -X 13 -X 14 -
X 15 -X 16 -X 17 ,
wherein;
X 1 is I, D, M, or T;
X 2 is I;
X 3 is S;
X 4 is Y, N, S, Q, V, T, H, or D;
X 5 is D;
X 6 is G;
X 7 is S, R, I, T, G, K, H, or N;
X 8 is N, Y, S, I, or T;
X 9 is K, M, N, Q, E, T, R, S, A, or L;
X 10 is Y, D, or E;
X 11 is S or Y;
X 12 is A;
X 13 is D;
X 14 is S;
X 15 is V;
X 16 is K; and
X 17 is G;
CDR-H3:
(SEQ ID NO: 107)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 ,
wherein;
X 1 is A;
X 2 is G, A, or R;
X 3 is G;
X 4 is G, S, or A;
X 5 is N;
X 6 is V or M;
X 7 is G;
X 8 is F, L, Y, or M;
X 9 is D; and
X 10 is I, S, or L;
CDR-L1:
(SEQ ID NO: 108)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 -X 11 ,
wherein;
X 1 is S;
X 2 is A or G;
X 3 is D;
X 4 is K, N, L, Q, M, E, S, T, G, or D;
X 5 is L;
X 6 is G;
X 7 is T, S, N, A, G, or E;
X 8 is K, Q, N, or R;
X 9 is Y;
X 10 is V or I; and
X 11 is S;
CDR-L2:
(SEQ ID NO: 109)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 ,
wherein;
X 1 is Q;
X 2 is D;
X 3 is A, G, W, S, or D;
X 4 is K, M, Q, N, L, T, I, or E;
X 5 is R;
X 6 is P; and
X 7 is S;
and
CDR-L3:
(SEQ ID NO: 110)
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 ,
wherein;
X 1 is Q;
X 2 is S or A;
X 3 is W;
X 4 is D;
X 5 is R, S, Q, P, A, V, W, or M;
X 6 is S, G, I, N, R, or T;
X 7 is D or G;
X 8 is V, A, P, or E; and
X 9 is V.
95 . The binding protein according to claim 94 , wherein said at least one CDR comprises an amino acid sequence selected from the group consisting of:
residues 31-35 of SEQ ID NO:112 (CDR-H1); residues 50-66 of SEQ ID NO:112 (CDR-H2); residues 99-108 of SEQ ID NO:112 (CDR-H3); residues 23-33 of SEQ ID NO:113 (CDR-L1); residues 49-55 of SEQ ID NO:113 (CDR-L2); residues 88-96 of SEQ ID NO:113 (CDR-L3); residues 31-35 of SEQ ID NO:316 (CDR-H1); residues 50-66 of SEQ ID NO:316 (CDR-H2); residues 99-108 of SEQ ID NO:316 (CDR-H3); residues 31-35 of SEQ ID NO:317 (CDR-H1); residues 50-66 of SEQ ID NO:317 (CDR-H2); residues 99-108 of SEQ ID NO:317 (CDR-H3); residues 31-35 of SEQ ID NO:318 (CDR-H1); residues 50-66 of SEQ ID NO:318 (CDR-H2); residues 99-108 of SEQ ID NO:318 (CDR-H3); residues 31-35 of SEQ ID NO:319 (CDR-H1); residues 50-66 of SEQ ID NO:319 (CDR-H2); residues 99-108 of SEQ ID NO:319 (CDR-H3); residues 31-35 of SEQ ID NO:320 (CDR-H1); residues 50-66 of SEQ ID NO:320 (CDR-H2); residues 99-108 of SEQ ID NO:320 (CDR-H3); residues 31-35 of SEQ ID NO:321 (CDR-H1); residues 50-66 of SEQ ID NO:321 (CDR-H2); residues 99-108 of SEQ ID NO:321 (CDR-H3); residues 31-35 of SEQ ID NO:322 (CDR-H1); residues 50-66 of SEQ ID NO:322 (CDR-H2); residues 99-108 of SEQ ID NO:322 (CDR-H3); residues 31-35 of SEQ ID NO:323 (CDR-H1); residues 50-66 of SEQ ID NO:323 (CDR-H2); residues 99-108 of SEQ ID NO:323 (CDR-H3); residues 31-35 of SEQ ID NO:324 (CDR-H1); residues 50-66 of SEQ ID NO:324 (CDR-H2); residues 99-108 of SEQ ID NO:324 (CDR-H3); residues 31-35 of SEQ ID NO:325 (CDR-H1); residues 50-66 of SEQ ID NO:325 (CDR-H2); residues 99-108 of SEQ ID NO:325 (CDR-H3); residues 31-35 of SEQ ID NO:326 (CDR-H1); residues 50-66 of SEQ ID NO:326 (CDR-H2); residues 99-108 of SEQ ID NO:326 (CDR-H3); residues 23-33 of SEQ ID NO:327 (CDR-L1); residues 49-55 of SEQ ID NO:327 (CDR-L2); residues 88-96 of SEQ ID NO:327 (CDR-L3); residues 23-33 of SEQ ID NO:328 (CDR-L1); residues 49-55 of SEQ ID NO:328 (CDR-L2); residues 88-96 of SEQ ID NO:328 (CDR-L3); residues 23-33 of SEQ ID NO:329 (CDR-L1); residues 49-55 of SEQ ID NO:329 (CDR-L2); residues 88-96 of SEQ ID NO:329 (CDR-L3); residues 23-33 of SEQ ID NO:330 (CDR-L1); residues 49-55 of SEQ ID NO:330 (CDR-L2); residues 88-96 of SEQ ID NO:330 (CDR-L3); residues 23-33 of SEQ ID NO:331 (CDR-L1); residues 49-55 of SEQ ID NO:331 (CDR-L2); residues 88-96 of SEQ ID NO:331 (CDR-L3); residues 23-33 of SEQ ID NO:332 (CDR-L1); residues 49-55 of SEQ ID NO:332 (CDR-L2); residues 88-96 of SEQ ID NO:332 (CDR-L3); residues 23-33 of SEQ ID NO:333 (CDR-L1); residues 49-55 of SEQ ID NO:333 (CDR-L2); residues 88-96 of SEQ ID NO:333 (CDR-L3).
96 . The binding protein according to claim 95 , wherein said binding protein comprises at least 3 CDRs.
97 . The binding protein according to claim 96 , wherein said at least 3 CDRs comprises a variable domain CDR set selected from the group consisting of:
VH A10 CDR Set CDR-H1: residues 31-35 of SEQ ID NO:112 CDR-H2: residues 50-66 of SEQ ID NO:112 CDR-H3: residues 99-108 of SEQ ID NO:112 VL A10 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:113 CDR-L2: residues 49-55 of SEQ ID NO:113 CDR-L3: residues 88-96 of SEQ ID NO:113 VH A10.3 CDR Set CDR-H1: residues 31-35 of SEQ ID NO:316 CDR-H2: residues 50-66 of SEQ ID NO:316 CDR-H3: residues 99-108 of SEQ ID NO:316 VL A10.3 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:327 CDR-L2: residues 49-55 of SEQ ID NO:327 CDR-L3: residues 88-96 of SEQ ID NO:327 VH A10.K30 CDR Set CDR-H1: residues 31-35 of SEQ ID NO:317 CDR-H2: residues 50-66 of SEQ ID NO:317 CDR-H3: residues 99-108 of SEQ ID NO:317 VH A10.K42 CDR Set CDR-H1: residues 31-35 of SEQ ID NO:318 CDR-H2: residues 50-66 of SEQ ID NO:318 CDR-H3: residues 99-108 of SEQ ID NO:318 VH A10.9A CDR Set CDR-H1: residues 31-35 of SEQ ID NO:319 CDR-H2: residues 50-66 of SEQ ID NO:319 CDR-H3: residues 99-108 of SEQ ID NO:319 VH A10.8A CDR Set CDR-H1: residues 31-35 of SEQ ID NO:320 CDR-H2: residues 50-66 of SEQ ID NO:320 CDR-H3: residues 99-108 of SEQ ID NO:320 VH A10.1A CDR Set CDR-H1: residues 31-35 of SEQ ID NO:321 CDR-H2: residues 50-66 of SEQ ID NO:321 CDR-H3: residues 99-108 of SEQ ID NO:321 VH A10.5D CDR Set CDR-H1: residues 31-35 of SEQ ID NO:322 CDR-H2: residues 50-66 of SEQ ID NO:322 CDR-H3: residues 99-108 of SEQ ID NO:322 VH A10.3A CDR Set CDR-H1: residues 31-35 of SEQ ID NO:323 CDR-H2: residues 50-66 of SEQ ID NO:323 CDR-H3: residues 99-108 of SEQ ID NO:323 VL A10.3A CDR Set CDR-L1: residues 23-33 of SEQ ID NO:330 CDR-L2: residues 49-55 of SEQ ID NO:330 CDR-L3: residues 88-96 of SEQ ID NO:330 VH A10.6B CDR Set CDR-H1: residues 31-35 of SEQ ID NO:324 CDR-H2: residues 50-66 of SEQ ID NO:324 CDR-H3: residues 99-108 of SEQ ID NO:324 VL A10.6B CDR Set CDR-L1: residues 23-33 of SEQ ID NO:331 CDR-L2: residues 49-55 of SEQ ID NO:331 CDR-L3: residues 88-96 of SEQ ID NO:331 VH A10.3D CDR Set CDR-H1: residues 31-35 of SEQ ID NO:325 CDR-H2: residues 50-66 of SEQ ID NO:325 CDR-H3: residues 99-108 of SEQ ID NO:325 VL A10.3D CDR Set CDR-L1: residues 23-33 of SEQ ID NO:332 CDR-L2: residues 49-55 of SEQ ID NO:332 CDR-L3: residues 88-96 of SEQ ID NO:332 VH A10.4C CDR Set CDR-H1: residues 31-35 of SEQ ID NO:326 CDR-H2: residues 50-66 of SEQ ID NO:326 CDR-H3: residues 99-108 of SEQ ID NO:326 VL A10.4C CDR Set CDR-L1: residues 23-33 of SEQ ID NO:333 CDR-L2: residues 49-55 of SEQ ID NO:333 CDR-L3: residues 88-96 of SEQ ID NO:333 VL A10.L45 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:328 CDR-L2: residues 49-55 of SEQ ID NO:328 CDR-L3: residues 88-96 of SEQ ID NO:328 VL A10.L73 CDR Set CDR-L1: residues 23-33 of SEQ ID NO:329 CDR-L2: residues 49-55 of SEQ ID NO:329 CDR-L3: residues 88-96 of SEQ ID NO:329
98 . The binding protein according to claim 97 , comprising at least two variable domain CDR Sets.
99 . The binding protein according to claim 98 , wherein said at least two variable domain CDR sets are selected from a group consisting of:
VH A10 CDR Set and VL A10 CDR Set; VH A10.3 CDR Set and VL A10.3 CDR Set; VH A10.3A CDR Set and VL A10.3A Set; VH A10.6B CDR Set and VL A10.6B Set; VH A10.3D CDR Set and VL A10.3D CDR Set; VH A10.4C CDR Set and VL A10.4C CDR Set; VH A10.K30 CDR Set and VL A10.3 CDR Set; VH A10.K42 CDR Set and VL A10.3 CDR Set; VH A10.3 CDR Set and VL A10.L45 CDR Set; VH A10.3 CDR Set and VL A10.L73 CDR Set; VH A10.9A CDR Set and VL A10.3 CDR Set; VH A10.8A CDR Set and VL A10.3 CDR Set; VH A10.1A CDR Set and VL A10.3 CDR Set; and VH A10.5D CDR Set and VL A10.3 CDR Set.
100 . The binding protein according to any one of claims 94 - 99 , further comprising a human acceptor framework.
101 . The binding protein according to claim 100 , wherein said human acceptor framework comprises an amino acid sequence selected from the group consisting of:
heavy chain acceptor framework sequences SEQ ID NOS:6-22, heavy chain acceptor sequences SEQ ID NOS:35-62, light chain acceptor sequences SEQ ID NOS:23-34, and light chain acceptor sequences SEQ ID NOS:63-98.
102 . The binding protein according to claim 100 or claim 101 , wherein said human acceptor framework comprises at least one framework region amino acid substitution, wherein the amino acid sequence of the framework is at least 65% identical to the sequence of said human acceptor framework and comprising at least 70 amino acid residues identical to said human acceptor framework.
103 . The binding protein according to claim 101 , wherein said human acceptor framework comprises at least one framework region amino acid substitution at a key residue, said key residue selected from the group consisting of:
a residue adjacent to a CDR; a glycosylation site residue; a rare residue; a residue capable of interacting with human DLL4 a canonical residue; a contact residue between heavy chain variable region and light chain variable region; a residue within a Vernier zone; and a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework.
104 . The binding protein according to claim 103 , wherein key residue selected from the group consisting of: 2H, 4H, 24H, 26H, 27H, 29H, 34H, 35H, 37H, 39H, 44H, 45H, 47H, 48H, 49H, 50H, 51H, 58H, 59H, 60H, 63H, 67H, 69H, 71H, 73H, 76H, 78H, 91H, 93H, 94H, 2L, 4L, 25L, 29L, 27bL, 33L, 34L, 36L, 38L, 43L, 44L, 46L, 47L, 48L, 49L, 55L, 58L, 62L, 64L, 71L, 87L, 89L, 90L, 91L, 94L, 95L.
105 . The binding protein according to claim 104 , where the binding protein is a consensus human variable domain.
106 . The binding protein according to claim 94 , where said binding protein comprises at least one variable domain having an amino acid sequence selected from the group consisting of:
SEQ ID NOS:112, 113, 316, 327, 317, 318, 319, 320, 321, 322, 323, 330, 324, 331, 325, 332, 326, 333, 328, and 329.
107 . The binding protein according to claim 106 , wherein said binding protein comprises two variable domains, wherein said two variable domains have amino acid sequences selected from the group consisting of: SEQ ID NOS:112 and 113, SEQ ID NOS:316 and 327, SEQ ID NOS:323 and 330, SEQ ID NOS:324 and 331, SEQ ID NOS:325 and 332, and SEQ ID NOS:326 and 333.
108 . The binding protein according to claim 104 , wherein said binding protein comprises at least one variable domain having an amino acid sequence selected from the group consisting of: SEQ ID NOS: 112, 113, 316, 327, 317, 318, 319, 320, 321, 322, 323, 330, 324, 331, 325, 332, 326, 333, 328, and 329.
109 . The binding protein according to claim 95 wherein said antigen binding domain comprises a V H .
110 . The binding protein according to claim 109 wherein said V H comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:112, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, and 326.
111 . The binding protein according to claim 95 wherein said antigen binding domain comprises a V L .
112 . The binding protein according to claim 111 wherein said V L comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:113, 327, 328, 329, 330, 331, 332, and 333.
113 . The binding protein according to claim 95 wherein said antigen binding domain comprises a V H and a V L .
114 . The binding protein according to claim 112 further comprising a V H wherein said V H comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS: SEQ ID NOS:112, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, and 326.
115 . The binding protein according to claim 20 wherein said V L comprises an amino acid sequence selected from the group consisting of:
SEQ ID NOS:113, 327, 328, 329, 330, 331, 332, and 333.
116 . The binding protein according to claim 95 , further comprising a heavy chain immunoglobulin constant domain selected from the group consisting of: a human IgM constant domain; a human IgG1 constant domain; a human IgG2 constant domain; a human IgG3 constant domain; a human IgG4 constant domain; a human IgE constant domain and a human IgA constant domain.
117 . The binding protein according to claim 117 wherein said heavy chain immunoglobulin constant region domain is a human IgG1 constant domain.
118 . The binding protein according to claim 117 wherein said human IgG1 constant domain comprises amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:3.
119 . The binding protein according to claim 95 , further comprising a light chain immunoglobulin constant domain selected from the group consisting of: a human Ig kappa constant domain and a human Ig lambda constant domain.
120 . The binding protein according to claim 119 wherein said light chain immunoglobulin constant region domain is a human Ig kappa constant domain comprising amino acid sequence SEQ ID NO:4.
121 . The binding protein according to claim 119 wherein said light chain immunoglobulin constant region domain is a human Ig lambda constant domain comprising amino acid sequence SEQ ID NO:5.
122 . The binding protein according to claim 94 wherein said binding protein is selected from the group consisting of: an immunoglobulin molecule, an scFv, a monoclonal antibody, a human antibody, a chimeric antibody, a humanized antibody, a single domain antibody, a Fab fragment, a Fab′ fragment, an F(ab′) 2 , an Fv, a disulfide linked Fv, a single domain antibody, a diabody, a multispecific antibody, a bispecific antibody, and a dual specific antibody.
123 . The binding protein according to claim 122 wherein said binding protein is a human antibody.
124 . A binding protein capable of binding human DLL-4, said binding protein comprising:
an Ig constant heavy region having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:3; an Ig constant light region having an amino acid sequence selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:5; an Ig variable heavy region having an amino acid sequence selected from the group consisting: SEQ ID NOS: SEQ ID NOS:112, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, and 326; and an Ig variable light region having an amino acid sequence selected from the group consisting: SEQ ID NOS:113, 327, 328, 329, 330, 331, 332, and 333.
125 . The binding protein according to claim 124 , wherein the Ig constant light region is SEQ ID NO:5.
126 . The binding protein according to any one of claims 94 - 125 , wherein the binding protein is capable of blocking DLL4 interaction with a Notch protein selected from the group consisting of Notch-1, Notch-2, Notch-3, Notch-4, and combinations thereof.
127 . The binding protein according to claim 126 , wherein the binding protein is capable of blocking DLL4 interaction with Notch-1 and Notch-4.
128 . The binding protein according to claim 126 , wherein the binding protein is capable of blocking DLL4 interaction with Notch-1.
129 . The binding protein according to claim 126 , wherein the binding protein is capable of blocking DLL4 interaction with Notch-4.
130 . The binding protein according to any one of claims 94 - 125 , wherein the binding protein is capable of modulating a biological function of DLL4.
131 . The binding protein according to any one of claims 94 - 125 , wherein said binding protein is capable of neutralizing a DLL4.
132 . The neutralizing binding protein according to claim 131 wherein said DLL4 is selected from the group consisting of: human DLL4, mouse DLL4, cynomolgus DLL4, and rat DLL4.
133 . The neutralizing binding protein according to claim 131 wherein said neutralizing binding protein diminishes the ability of DLL4 to bind to its receptor.
134 . The neutralizing binding protein according to claim 131 wherein said neutralizing binding protein is capable of reducing normal angiogenesis.
135 . The neutralizing binding protein according to claim 131 , wherein said neutralizing binding protein has a dissociation constant (K D ) selected from the group consisting of: at most about 10 −7 M; at most about 10 −8 M; at most about 10 −9 M; at most about 10 −10 M; at most about 10 −11 M; at most about 10 −12 M; and at most 10 −13 M.
136 . The neutralizing binding protein according to claim 131 , wherein said neutralizing binding protein has an on rate selected from the group consisting of: at least about 10 2 M −1 s −1 ; at least about 10 3 M −1 s −1 ; at least about 10 4 M −1 s −1 ; at least about 10 −1 M −1 s −1 ; and at least about 10 6 M −1 s −1 .
137 . The neutralizing binding protein according to claim 131 , wherein said neutralizing binding protein has an off rate selected from the group consisting of: at most about 10 −3 s −1 ; at most about 10 −4 s −1 ; at most about 10 −5 s −1 ; and at most about 10 −6 s −1 .
138 . A labeled binding protein comprising a binding protein of any one of claims 94 - 137 , wherein said binding protein is conjugated to a detectable label.
139 . The labeled binding protein of claim 138 , wherein the detectable label is selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
140 . The labeled binding protein of claim 139 , wherein said label is a radiolabel selected from the group consisting of: 3 H, 14 C, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho, and 153 Sm.
141 . An antibody construct comprising a binding protein described in any of claims 94 - 137 and further comprising a linker polypeptide or an immunoglobulin constant domain.
142 . The antibody construct according to claim 141 , selected from the group consisting of:
an immunoglobulin molecules, a monoclonal antibody, a chimeric antibody, a CDR-grafted antibody, a humanized antibody, a Fab, a Fab′, a F(ab′) 2 , a Fv, a disulfide linked Fv, a scFv, a single domain antibody, a diabody, a multispecific antibody, a dual specific antibody, and a bispecific antibody.
143 . The antibody construct according to claim 141 , wherein said antibody construct comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:
a human IgM constant domain, a human IgG1 constant domain, a human IgG2 constant domain, a human IgG3 constant domain, a human IgG4 constant domain, a human IgE constant domain, a human IgA constant domain, and a IgG constant domain variant with one or more mutations altering binding strength to Fc neonatal receptor, Fc gamma receptors, or C1q.
144 . The antibody construct according to claim 141 , comprising an immunoglobulin constant domain having an amino acid sequence selected from consisting of: SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and combinations thereof.
145 . An antibody conjugate comprising an antibody construct as described in claim 141 , wherein said antibody construct is conjugated to a therapeutic or cytotoxic agent.
146 . The antibody conjugate of claim 145 , wherein said therapeutic or cytotoxic agent is selected from the group consisting of: an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.
147 . An isolated nucleic acid encoding a polypeptide selected from the group consisting of: a polypeptide comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises one or more of CDR-H1, a CDR-H2, or a CDR-H3 as described in claim 94 ; a polypeptide comprising a light chain variable domain, wherein the light chain variable domain comprises one or more of CDR-L1, a CDR-L2, or a CDR-L3 as described in claim 94 ; or a combination of both polypeptides.
148 . A vector comprising an isolated nucleic acid according to claim 147 .
149 . The vector of claim 148 wherein said vector is selected from the group consisting of: pcDNA, pTT, pTT3, pEFBOS, pBV, pJV, pHybE and pBJ.
150 . A host cell comprising a vector according to any one of claims 148 and 149 .
151 . The host cell according to claim 150 , wherein said host cell is a prokaryotic cell.
152 . The host cell according to claim 151 , wherein said host cell is Escherichia coli.
153 . The host cell according to claim 152 , wherein said host cell is a eukaryotic cell.
154 . The host cell according to claim 153 , wherein said eukaryotic cell is selected from the group consisting of: a protist cell, an animal cell, a plant cell, and a fungal cell.
155 . The host cell according to claim 154 , wherein said eukaryotic cell is an animal cell selected from the group consisting of: a mammalian cell, an avian cell, and an insect cell.
156 . The host cell according to claim 155 , wherein said mammalian cell is a CHO cell.
157 . The host cell according to claim 155 , wherein said mammalian cell is a COS cell.
158 . The host cell according to claim 154 , wherein said fungal cell is Saccharomyces cerevisiae.
159 . The host cell according to claim 155 , wherein said insect cell is an Sf9 cell.
160 . A method of producing a binding protein that binds human DLL4, comprising culturing the host cell of any one of claims 150 - 159 in a culture medium under conditions sufficient to produce a binding protein that binds human DLL4.
161 . A binding protein produced according to the method of claim 160 .
162 . A crystallized binding protein comprising a binding protein according to any one of claims 94 - 137 , wherein said binding protein exists as a crystal.
163 . The crystallized binding protein according to claim 162 , wherein said crystal is a carrier-free pharmaceutical controlled release crystal.
164 . The crystallized binding protein according to claim 162 , wherein said binding protein has a greater half life in vivo than the soluble counterpart of said binding protein.
165 . The crystallized binding protein according to claim 162 , wherein said binding protein retains biological activity.
166 . A composition for the release of a binding protein said composition comprising:
(a) a formulation, wherein said formulation comprises a crystallized binding protein, according to any one of claims 162 - 165 , and an ingredient; and (b) at least one polymeric carrier.
167 . The composition according to claim 166 , wherein said polymeric carrier is a polymer selected from one or more of the group consisting of poly (acrylic acid), poly (cyanoacrylates), poly (amino acids), poly (anhydrides), poly (depsipeptide), poly (esters), poly (lactic acid), poly (lactic-co-glycolic acid) or PLGA, poly (b-hydroxybutryate), poly (caprolactone), poly (dioxanone), poly (ethylene glycol), poly ((hydroxypropyl) methacrylamide, poly [(organo)phosphazene], poly (ortho esters), poly (vinyl alcohol), poly (vinylpyrrolidone), maleic anhydride-alkyl vinyl ether copolymers, pluronic polyols, albumin, alginate, cellulose and cellulose derivatives, collagen, fibrin, gelatin, hyaluronic acid, oligosaccharides, glycaminoglycans, sulfated polyeaccharides, blends and copolymers thereof.
168 . The composition according to claim 166 , wherein said ingredient is selected from the group consisting of albumin, sucrose, trehalose, lactitol, gelatin, hydroxypropyl-β-cyclodextrin, methoxypolyethylene glycol and polyethylene glycol.
169 . A method for treating a mammal comprising the step of administering to the mammal an effective amount of the composition according to claim 166 .
170 . A pharmaceutical composition comprising the binding protein of any one of claims 94 - 137 , and a pharmaceutically acceptable carrier.
172 . The pharmaceutical composition of claim 170 which further comprises at least one additional therapeutic agent for treating a disorder in which DLL4 activity is detrimental.
173 . The pharmaceutical composition of claim 172 , wherein said additional agent is selected from the group consisting of: angiogenesis inhibitors; kinase inhibitors; co-stimulation molecule blockers; adhesion molecule blockers; anti-cytokine antibody or functional fragment thereof; methotrexate; corticosteroids; cyclosporine; rapamycin; FK506; and non-steroidal anti-inflammatory agents.
174 . A method for reducing human DLL4 activity comprising contacting human DLL4 with the binding protein of any one of claims 94 - 137 such that human DLL4 activity is reduced.
175 . A method for reducing human DLL4 activity in a human subject suffering from a disorder in which DLL4 activity is detrimental, comprising administering to the human subject the binding protein of any one of claims 94 - 137 such that human IL-17 activity in the human subject is reduced.
176 . A method for treating a subject for a disease or a disorder in which DLL4 activity is detrimental by administering to the subject the binding protein of any one of claims 94 - 137 such that treatment is achieved.
177 . The method of claim 176 , wherein said disorder is selected from the group consisting of: breast cancer, colon cancer, rectal cancer, lung cancer, oropharynx cancer, hypopharynx cancer, esophageal cancer, stomach cancer, pancreas cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, female genital tract cancer, male genital tract cancer, endocrine gland cancer, skin cancer, hemangiomas, melanomas, sarcomas, brain tumor, nerve cancer, eye tumor, meninges cancer, solid tumors from hematopoietic malignancies, tumor metastases, ocular neovascularization, edema, rheumatoid arthritis, multiple sclerosis, atheroscleorotic plaques, Crohn's disease, inflammatory bowel disease, refractory ascites, psoriasis, sarcoidosis, arterial arteriosclerosis, sepsis, peptic ulcers, burns, and pancreatitis, polycystic ovarian disease (POD), endometriosis, uterine fibroids, benign prostate hypertrophy, and other angiogenesis independent and dependent diseases characterized by abberant DLL4 activity.
178 . The method according to claim 177 , wherein the disorder is a primary and metastatic cancer.
179 . The method according to claim 177 , wherein the urinary tract cancer is selected from the group consisting of renal cancer, bladder cancer, and urothelium cancer.
180 . The method according to claim 177 , wherein the female genital tract cancer is selected from the group consisting of cervical cancer, uterine cancer, ovarian cancer, choriocarcinoma, and gestational trophoblastic disease.
181 . The method according to claim 177 , wherein the male genital tract cancer is selected from the group consisting of prostate cancer, seminal vesicles cancer, testicular cancer, and germ cell tumor.
182 . The method according to claim 177 , wherein the endocrine gland cancer is selected from the group consisting of thyroid cancer, adrenal cancer, and pituitary gland cancer.
183 . The method according to claim 177 , wherein the sarcoma is selected from the group consisting of a bone sarcoma, a soft tissue sarcoma, and Kaposi's sarcoma.
184 . The method according to claim 177 , wherein the meninges cancer is selected from the group consisting of an astrocytoma, a glioma, a glioblastoma, a retinoblastoma, a neuroma, a neuroblastoma, a Schwannoma, and a meningiomas.
185 . The method according to claim 177 , wherein the solid tumor from a hematopoietic malignancy is a leukemia, a Hodgkin's leukemia, a non-Hodgkin's leukemia, a lymphoma, a Hodgkin's lymphoma, and a non-Hodgkin's lymphomas.
186 . The method according to claim 177 , wherein the ocular neovascularization is selected from the group consisting of diabetic blindness, a retinopathy, an age-induced macular degeneration, and a rubeosis.
187 . A method of treating a patient suffering from a disorder in which DLL4 is detrimental comprising the step of administering the binding protein of any one of claims 94 - 137 before, concurrent with, or after the administration of a second agent, wherein the second agent is selected from the group consisting of an antibody or fragment thereof capable of binding human VEGFR2; methotrexate; an antibody, or fragment thereof, capable of binding human TNF; corticosteroids, cyclosporine, rapamycin, FK506, and non-steroidal anti-inflammatory agents.Join the waitlist — get patent alerts
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