Therapeutic DLL4 Binding Proteins
DLL4 binding proteins are described herein, including antibodies, CDR-grafted antibodies, humanized antibodies, and DLL4 binding fragments thereof, proteins that bind DLL4 with high affinity, and DLL4 binding proteins that neutralize DLL4 and/or VEGF activity. The DLL4 binding proteins are useful for treating or preventing cancers and tumors and especially for treating or preventing tumor angiogenesis.
1 . A binding protein capable of binding DLL4, comprising at least one amino acid sequence selected from the group of amino acid sequences consisting of SEQ ID NO:157, SEQ ID NO:158, SEQ ID NO:159, SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:163, and SEQ ID NO:164.
2 . A binding protein comprising an antigen binding domain wherein the binding protein is capable of binding human DLL4, said antigen binding domain comprising at least one CDR, wherein:
CDR-H1 is selected from the group consisting of:
X 1 -X 2 -X 3 -X 4 -X 5 (SEQ ID NO:151), wherein;
X 1 is N, H, or Y;
X 2 is F;
X 3 is P;
X 4 is M; and
X 5 is A or S;
residues 31-35 of SEQ ID NO:157 (CDR-H1 38H12);
residues 31-35 of SEQ ID NO:161 (CDR-H1 37D10);
residues 31-35 of SEQ ID NO:163 (CDR-H1 32C7);
residues 31-35 of SEQ ID NO:165 (CDR-H1 14G1);
residues 31-35 of SEQ ID NO:167 (CDR-H1 14A11);
residues 31-35 of SEQ ID NO:169 (CDR-H1 15D6);
residues 31-35 of SEQ ID NO:171 (CDR-H1 VH.1 1A11);
residues 31-35 of SEQ ID NO:172 (CDR-H1 VH.1a 1A11);
residues 31-35 of SEQ ID NO:173 (CDR-H1 VH.1b 1A11);
residues 31-35 of SEQ ID NO:174 (CDR-H1 VH.2a 1A11);
residues 31-35 of SEQ ID NO:179 (CDR-H1 VH.1 38H12);
residues 31-35 of SEQ ID NO:180 (CDR-H1 VH.1A 38H12);
residues 31-35 of SEQ ID NO:181 (CDR-H1 VH.1b 38H12);
residues 31-35 of SEQ ID NO:182 (CDR-H1 VH.2a 38H12);
residues 31-35 of SEQ ID NO:187 (CDR-H1 h1A11VH.1);
residues 31-35 of SEQ ID NO:188 (CDR-H1 h1A11.A6);
residues 31-35 of SEQ ID NO:189 (CDR-H1 h1A11.A8);
residues 31-35 of SEQ ID NO:190 (CDR-H1 h1A11.C6);
residues 31-35 of SEQ ID NO:191 (CDR-H1 h1A11.A11);
residues 31-35 of SEQ ID NO:192 (CDR-H1 h1A11.B5);
residues 31-35 of SEQ ID NO:193 (CDR-H1 h1A11.E12);
residues 31-35 of SEQ ID NO:194 (CDR-H1 h1A11.G3);
residues 31-35 of SEQ ID NO:195 (CDR-H1 h1A11.F5); and
residues 31-35 of SEQ ID NO:196 (CDR-H1 h1A11.H2);
CDR-H2 is selected from the group consisting of:
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 (SEQ ID NO:152), wherein;
X 1 is T or S;
X 2 is I;
X 3 is S;
X 4 is S or G;
X 5 is S;
X 6 is D;
X 7 is G, A, D, S, or E;
X 8 is T or W;
X 9 is T, P, or A;
X 10 is Y, S, T, or N;
X 11 is Y or I;
X 12 is R or G;
X 13 is D;
X 14 is 5;
X 15 is V;
X 16 is K; and
X 17 is G;
residues 50-66 of SEQ ID NO:157 (CDR-H2 38H12);
residues 50-68 of SEQ ID NO:161 (CDR-H2 37D10);
residues 50-66 of SEQ ID NO:163 (CDR-H2 32C7);
residues 50-66 of SEQ ID NO:165 (CDR-H2 14G1);
residues 50-66 of SEQ ID NO:167 (CDR-H2 14A11);
residues 50-66 of SEQ ID NO:169 (CDR-H2 15D6);
residues 50-66 of SEQ ID NO:171 (CDR-H2 VH.1 1A11);
residues 50-66 of SEQ ID NO:172 (CDR-H2 VH.1a 1A11);
residues 50-66 of SEQ ID NO:173 (CDR-H2 VH.1b 1A11);
residues 50-66 of SEQ ID NO:174 (CDR-H2 VH.2a 1A11);
residues 50-66 of SEQ ID NO:179 (CDR-H2 VH.1 38H12);
residues 50-66 of SEQ ID NO:180 (CDR-H2 VH.1A 38H12);
residues 50-66 of SEQ ID NO:181 (CDR-H2 VH.1b 38H12);
residues 31-35 of SEQ ID NO:182 (CDR-H1 VH.2a 38H12);
residues 50-66 of SEQ ID NO:187 (CDR-H2 h1A11VH.1);
residues 50-66 of SEQ ID NO:188 (CDR-H2 h1A11.A6);
residues 50-66 of SEQ ID NO:189 (CDR-H2 h1A11.A8);
residues 50-66 of SEQ ID NO:190 (CDR-H2 h1A11.C6);
residues 50-66 of SEQ ID NO:191 (CDR-H2 h1A11.A11);
residues 50-66 of SEQ ID NO:192 (CDR-H2 h1A11.B5);
residues 50-66 of SEQ ID NO:193 (CDR-H2 h1A11.E12);
residues 50-66 of SEQ ID NO:194 (CDR-H2 h1A11.G3);
residues 50-66 of SEQ ID NO:195 (CDR-H2 h1A11.F5); and
residues 50-66 of SEQ ID NO:196 (CDR-H2 h1A11.H2);
CDR-H3 is selected from the group consisting of:
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 (SEQ ID NO:153), wherein;
X 1 is G;
X 2 is Y;
X 3 is Y;
X 4 is N;
X 5 is S;
X 6 is P;
X 7 is F;
X 8 is A; and
X 9 is Y, F, or S;
residues 99-107 of SEQ ID NO:157 (CDR-H3 38H12);
residues 101-111 of SEQ ID NO:161 (CDR-H3 37D10);
residues 99-105 of SEQ ID NO:163 (CDR-H3 32C7);
residues 99-105 of SEQ ID NO:165 (CDR-H3 14G1);
residues 99-110 of SEQ ID NO:167 (CDR-H3 14A11);
residues 99-110 of SEQ ID NO:169 (CDR-H3 15D6);
residues 99-107 of SEQ ID NO:171 (CDR-H3 VH.1 1A11);
residues 99-107 of SEQ ID NO:172 (CDR-H3 VH.1a 1A11);
residues 99-107 of SEQ ID NO:173 (CDR-H3 VH.1b 1A11);
residues 99-107 of SEQ ID NO:174 (CDR-H3 VH.2a 1A11);
residues 99-107 of SEQ ID NO:179 (CDR-H3 VH.1 38H12);
residues 99-107 of SEQ ID NO:180 (CDR-H3 VH.1A 38H12);
residues 99-107 of SEQ ID NO:181 (CDR-H2 VH.1b 38H12);
residues 99-107 of SEQ ID NO:182 (CDR-H1 VH.2a 38H12);
residues 99-107 of SEQ ID NO:187 (CDR-H3 h1A11VH.1);
residues 99-107 of SEQ ID NO:188 (CDR-H3 h1A11.A6);
residues 99-107 of SEQ ID NO:189 (CDR-H3 h1A11.A8);
residues 99-107 of SEQ ID NO:190 (CDR-H3 h1A11.C6);
residues 99-107 of SEQ ID NO:191 (CDR-H3 h1A11.A11);
residues 99-107 of SEQ ID NO:192 (CDR-H3 h1A11.B5);
residues 99-107 of SEQ ID NO:193 (CDR-H3 h1A11.E12);
residues 99-107 of SEQ ID NO:194 (CDR-H3 h1A11.G3);
residues 99-107 of SEQ ID NO:195 (CDR-H3 h1A11.F5); and
residues 99-107 of SEQ ID NO:196 (CDR-H3 h1A11.H2);
CDR-L1 is selected from the group consisting of:
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 -X 10 -X 11 (SEQ ID NO:154), wherein;
X 1 is R;
X 2 is A;
X 3 is S;
X 4 is E or Q;
X 5 is D or E;
X 6 is I;
X 7 is Y or W;
X 8 is S, I, Y, N, or R;
X 9 is N;
X 10 is L; and
X 11 is A;
residues 24-34 of SEQ ID NO:158 (CDR-L1 38H12);
residues 24-34 of SEQ ID NO:162 (CDR-L1 37D10);
residues 24-34 of SEQ ID NO:164 (CDR-L1 32C7);
residues 24-34 of SEQ ID NO:166 (CDR-L1 14G1);
residues 23-37 of SEQ ID NO:168 (CDR-L1 14A11);
residues 23-37 of SEQ ID NO:170 (CDR-L1 15D6);
residues 24-34 of SEQ ID NO:175 (CDR-L1 VL.1 1A11);
residues 24-34 of SEQ ID NO:176 (CDR-L1 VL.1a 1A11);
residues 24-34 of SEQ ID NO:177 (CDR-L1 VL.1b 1A11);
residues 24-34 of SEQ ID NO:178 (CDR-L1 VL.2a 1A11);
residues 24-34 of SEQ ID NO:183 (CDR-L1 VL.1 38H12);
residues 24-34 of SEQ ID NO:184 (CDR-L1 VL.1a 38H12);
residues 24-34 of SEQ ID NO:185 (CDR-L1 VL.1b 38H12);
residues 24-34 of SEQ ID NO:186 (CDR-L1 VL.2a 38H12);
residues 24-34 of SEQ ID NO:197 (CDR-L1 h1A11VL.1);
residues 24-34 of SEQ ID NO:198 (CDR-L1 h1A11.A2);
residues 24-34 of SEQ ID NO:199 (CDR-L1 h1A11.A12);
residues 24-34 of SEQ ID NO:200 (CDR-L1 h1A11.A7);
residues 24-34 of SEQ ID NO:201 (CDR-L1 h1A11.B4);
residues 24-34 of SEQ ID NO:202 (CDR-L1 h1A11.B5); and
residues 24-34 of SEQ ID NO:203 (CDR-L1 h1A11.E12);
CDR-L2 is selected from group consisting of:
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 (SEQ ID NO:155), wherein;
X 1 is D;
X 2 is T;
X 3 is N or S;
X 4 is N, D, S, I, Y, or V;
X 5 is L;
X 6 is A; and
X 7 is D;
residues 50-56 of SEQ ID NO:158 (CDR-L2 38H12);
residues 50-56 of SEQ ID NO:162 (CDR-L2 37D10);
residues 50-56 of SEQ ID NO:164 (CDR-L2 32C7);
residues 50-56 of SEQ ID NO:166 (CDR-L2 14G1);
residues 53-59 of SEQ ID NO:168 (CDR-L2 14A11);
residues 53-59 of SEQ ID NO:170 (CDR-L2 15D6);
residues 50-56 of SEQ ID NO:175 (CDR-L2 VL.1 1A11);
residues 50-56 of SEQ ID NO:176 (CDR-L2 VL.1a 1A11);
residues 50-56 of SEQ ID NO:177 (CDR-L2 VL.1b 1A11);
residues 50-56 of SEQ ID NO:178 (CDR-L2 VL.2a 1A11);
residues 50-56 of SEQ ID NO:183 (CDR-L2 VL.1 38H12);
residues 50-56 of SEQ ID NO:184 (CDR-L2 VL.1a 38H12);
residues 50-56 of SEQ ID NO:185 (CDR-L2 VL.1b 38H12);
residues 50-56 of SEQ ID NO:186 (CDR-L2 VL.2a 38H12);
residues 50-56 of SEQ ID NO:197 (CDR-L2 h1A11VL.1);
residues 50-56 of SEQ ID NO:198 (CDR-L2 h1A11.A2);
residues 50-56 of SEQ ID NO:199 (CDR-L2 h1A11.A12);
residues 50-56 of SEQ ID NO:200 (CDR-L2 h1A11.A7);
residues 50-56 of SEQ ID NO:201 (CDR-L2 h1A11.B4);
residues 50-56 of SEQ ID NO:202 (CDR-L2 h1A11.B5); and
residues 50-56 of SEQ ID NO:203 (CDR-L2 h1A11.E12);
and
CDR-L3 is selected from the group consisting of:
X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -X 9 (SEQ ID NO:156), wherein;
X 1 is Q;
X 2 is Q;
X 3 is Y;
X 4 is N, D, or T;
X 5 is N, Y, or W;
X 6 is Y or V;
X 7 is P;
X 8 is P; and
X 9 is T.
residues 89-97 of SEQ ID NO:158 (CDR-L3 38H12);
residues 89-97 of SEQ ID NO:162 (CDR-L3 37D10);
residues 89-97 of SEQ ID NO:164 (CDR-L3 32C7);
residues 89-98 of SEQ ID NO:166 (CDR-L3 14G1);
residues 92-100 of SEQ ID NO:168 (CDR-L3 14A11);
residues 92-100 of SEQ ID NO:170 (CDR-L3 15D6);
residues 89-97 of SEQ ID NO:175 (CDR-L3 VL.1 1A11);
residues 89-97 of SEQ ID NO:176 (CDR-L3 VL.1a 1A11);
residues 89-97 of SEQ ID NO:177 (CDR-L3 VL.1b 1A11);
residues 89-97 of SEQ ID NO:178 (CDR-L3 VL.2a 1A11);
residues 89-97 of SEQ ID NO:183 (CDR-L3 VL.1 38H12);
residues 89-97 of SEQ ID NO:184 (CDR-L3 VL.1a 38H12);
residues 89-97 of SEQ ID NO:185 (CDR-L3 VL.1b 38H12);
residues 89-97 of SEQ ID NO:186 (CDR-L3 VL.2a 38H12);
residues 89-97 of SEQ ID NO:197 (CDR-L3 h1A11VL.1);
residues 89-97 of SEQ ID NO:198 (CDR-L3 h1A11.A2);
residues 89-97 of SEQ ID NO:199 (CDR-L3 h1A11.A12);
residues 89-97 of SEQ ID NO:200 (CDR-L3 h1A11.A7);
residues 89-97 of SEQ ID NO:201 (CDR-L3 h1A11.B4);
residues 89-97 of SEQ ID NO:202 (CDR-L3 h1A11.B5); and
residues 89-97 of SEQ ID NO:203 (CDR-L3 h1A11.E12).
3 . The binding protein according to claim 2 , wherein said at least one CDR comprises an amino acid sequence selected from the group consisting of:
residues 31-35 of SEQ ID NO:157 (CDR-H1 38H12); residues 50-66 of SEQ ID NO:157 (CDR-H2 38H12); residues 99-107 of SEQ ID NO:157 (CDR-H3 38H12); residues 24-34 of SEQ ID NO:158 (CDR-L1 38H12); residues 50-56 of SEQ ID NO:158 (CDR-L2 38H12); residues 89-97 of SEQ ID NO:158 (CDR-L3 38H12); residues 31-35 of SEQ ID NO:159 (CDR-H1 1A11); residues 50-66 of SEQ ID NO:159 (CDR-H2 1A11); residues 99-107 of SEQ ID NO:159 (CDR-H3 1A11); residues 24-34 of SEQ ID NO:160 (CDR-L1 1A11); residues 50-56 of SEQ ID NO:160 (CDR-L2 1A11); residues 89-97 of SEQ ID NO:160 (CDR-L3 1A11); residues 31-35 of SEQ ID NO:161 (CDR-H1 37D10); residues 50-68 of SEQ ID NO:161 (CDR-H2 37D10); residues 101-111 of SEQ ID NO:161 (CDR-H3 37D10);
residues 24-34 of SEQ ID NO:162 (CDR-L1 37D10); residues 50-56 of SEQ ID NO:162 (CDR-L2 37D10); residues 89-97 of SEQ ID NO:162 (CDR-L3 37D10); residues 31-35 of SEQ ID NO:163 (CDR-H1 32C7); residues 50-66 of SEQ ID NO:163 (CDR-H2 32C7); residues 99-105 of SEQ ID NO:163 (CDR-H3 32C7); residues 24-34 of SEQ ID NO:164 (CDR-L1 32C7); residues 50-56 of SEQ ID NO:164 (CDR-L2 32C7); residues 89-98 of SEQ ID NO:164 (CDR-L3 32C7); residues 31-35 of SEQ ID NO:165 (CDR-H1 14G1); residues 50-66 of SEQ ID NO:165 (CDR-H2 14G1); residues 99-105 of SEQ ID NO:165 (CDR-H3 14G1); residues 24-34 of SEQ ID NO:166 (CDR-L1 14G1); residues 50-56 of SEQ ID NO:166 (CDR-L2 14G1); residues 89-98 of SEQ ID NO:166 (CDR-L3 14G1); residues 31-35 of SEQ ID NO:167 (CDR-H1 14A11); residues 50-66 of SEQ ID NO:167 (CDR-H2 14A11); residues 99-110 of SEQ ID NO:167 (CDR-H3 14A11); residues 23-37 of SEQ ID NO:168 (CDR-L1 14A11); residues 53-59 of SEQ ID NO:168 (CDR-L2 14A11); residues 92-100 of SEQ ID NO:168 (CDR-L3 14A11); residues 31-35 of SEQ ID NO:169 (CDR-H1 15D6); residues 50-66 of SEQ ID NO:169 (CDR-H2 15D6); residues 99-110 of SEQ ID NO:169 (CDR-H3 15D6); residues 23-37 of SEQ ID NO:170 (CDR-L1 15D6); residues 53-59 of SEQ ID NO:170 (CDR-L2 15D6); residues 92-100 of SEQ ID NO:170 (CDR-L3 15D6); residues 31-35 of SEQ ID NO:171 (CDR-H1 VH.1 1A11); residues 50-66 of SEQ ID NO:171 (CDR-H2 VH.1 1A11); residues 99-107 of SEQ ID NO:171 (CDR-H3 VH.1 1A11); residues 31-35 of SEQ ID NO:172 (CDR-H1 VH.1a 1A11); residues 50-66 of SEQ ID NO:172 (CDR-H2 VH.1a 1A11); residues 99-107 of SEQ ID NO:172 (CDR-H3 VH.1a 1A11); residues 31-35 of SEQ ID NO:173 (CDR-H1 VH.1b 1A11); residues 50-66 of SEQ ID NO:173 (CDR-H2 VH.1b 1A11); residues 99-107 of SEQ ID NO:173 (CDR-H3 VH.1b 1A11); residues 31-35 of SEQ ID NO:174 (CDR-H1 VH.2a 1A11); residues 50-66 of SEQ ID NO:174 (CDR-H2 VH.2a 1A11); residues 99-107 of SEQ ID NO:174 (CDR-H3 VH.2a 1A11); residues 24-34 of SEQ ID NO:175 (CDR-L1 VL.1 1A11); residues 50-56 of SEQ ID NO:175 (CDR-L2 VL.1 1A11); residues 89-97 of SEQ ID NO:175 (CDR-L3 VL.1 1A11); residues 24-34 of SEQ ID NO:176 (CDR-L1 VL.1a 1A11); residues 50-56 of SEQ ID NO:176 (CDR-L2 VL.1a 1A11); residues 89-97 of SEQ ID NO:176 (CDR-L3 VL.1a 1A11); residues 24-34 of SEQ ID NO:177 (CDR-L1 VL.1b 1A11); residues 50-56 of SEQ ID NO:177 (CDR-L2 VL.1b 1A11); residues 89-97 of SEQ ID NO:177 (CDR-L3 VL.1b 1A11); residues 24-34 of SEQ ID NO:178 (CDR-L1 VL.2a 1A11); residues 50-56 of SEQ ID NO:178 (CDR-L2 VL.2a 1A11); residues 89-97 of SEQ ID NO:178 (CDR-L3 VL.2a 1A11); residues 31-35 of SEQ ID NO:179 (CDR-H1 VH.1 38H12); residues 50-66 of SEQ ID NO:179 (CDR-H2 VH.1 38H12); residues 99-107 of SEQ ID NO:179 (CDR-H3 VH.1 38H12); residues 31-35 of SEQ ID NO:180 (CDR-H1 VH.1A 38H12); residues 50-66 of SEQ ID NO:180 (CDR-H2 VH.1A 38H12); residues 99-107 of SEQ ID NO:180 (CDR-H3 VH.1A 38H12); residues 31-35 of SEQ ID NO:181 (CDR-H1 VH.1b 38H12); residues 50-66 of SEQ ID NO:181 (CDR-H2 VH.1b 38H12); residues 99-107 of SEQ ID NO:181 (CDR-H3 VH.1b 38H12); residues 31-35 of SEQ ID NO:182 (CDR-H1 VH.2a 38H12); residues 50-66 of SEQ ID NO:182 (CDR-H2 VH.2a 38H12); residues 99-107 of SEQ ID NO:182 (CDR-H3 VH.2a 38H12); residues 24-34 of SEQ ID NO:183 (CDR-L1 VL.1 38H12); residues 50-56 of SEQ ID NO:183 (CDR-L2 VL.1 38H12); residues 89-97 of SEQ ID NO:183 (CDR-L3 VL.1 38H12); residues 24-34 of SEQ ID NO:184 (CDR-L1 VL.1a 38H12); residues 50-56 of SEQ ID NO:184 (CDR-L2 VL.1a 38H12); residues 89-97 of SEQ ID NO:184 (CDR-L3 VL.1a 38H12); residues 24-34 of SEQ ID NO:185 (CDR-L1 VL.1b 38H12); residues 50-56 of SEQ ID NO:185 (CDR-L2 VL.1b 38H12); residues 89-97 of SEQ ID NO:185 (CDR-L3 VL.1b 38H12); residues 24-34 of SEQ ID NO:186 (CDR-L1 VL.2a 38H12); residues 50-56 of SEQ ID NO:186 (CDR-L2 VL.2a 38H12); residues 89-97 of SEQ ID NO:186 (CDR-L3 VL.2a 38H12); residues 31-35 of SEQ ID NO:187 (CDR-H1 h1A11VH.1), residues 50-66 of SEQ ID NO:187 (CDR-H2 h1A11VH.1); residues 99-107 of SEQ ID NO:187 (CDR-H3 h1A11VH.1); residues 31-35 of SEQ ID NO:188 (CDR-H1 h1A11.A6), residues 50-66 of SEQ ID NO:188 (CDR-H2 h1A11.A6); residues 99-107 of SEQ ID NO:188 (CDR-H3 h1A11.A6); residues 31-35 of SEQ ID NO:189 (CDR-H1 h1A11.A8), residues 50-66 of SEQ ID NO:189 (CDR-H2 h1A11.A8); residues 99-107 of SEQ ID NO:189 (CDR-H3 h1A11.A8); residues 31-35 of SEQ ID NO:190 (CDR-H1 h1A11.C6), residues 50-66 of SEQ ID NO:190 (CDR-H2 h1A11.C6); residues 99-107 of SEQ ID NO:190 (CDR-H3 h1A11.C6); residues 31-35 of SEQ ID NO:191 (CDR-H1 h1A11.A11), residues 50-66 of SEQ ID NO:191 (CDR-H2 h1A11.A11); residues 99-107 of SEQ ID NO:191 (CDR-H3 h1A11.A11); residues 31-35 of SEQ ID NO:192 (CDR-H1 h1A11.B5), residues 50-66 of SEQ ID NO:192 (CDR-H2 h1A11.B5); residues 99-107 of SEQ ID NO:192 (CDR-H3 h1A11.B5); residues 31-35 of SEQ ID NO:193 (CDR-H1 h1A11.E12), residues 50-66 of SEQ ID NO:193 (CDR-H2 h1A11.E12); residues 99-107 of SEQ ID NO:193 (CDR-H3 h1A11.E12); residues 31-35 of SEQ ID NO:194 (CDR-H1 h1A11.G3), residues 50-66 of SEQ ID NO:194 (CDR-H2 h1A11.G3); residues 99-107 of SEQ ID NO:194 (CDR-H3 h1A11.G3); residues 31-35 of SEQ ID NO:195 (CDR-H1 h1A11.F5), residues 50-66 of SEQ ID NO:195 (CDR-H2 h1A11.F5); residues 99-107 of SEQ ID NO:195 (CDR-H3 h1A11.F5); residues 31-35 of SEQ ID NO:196 (CDR-H1 h1A11.H2), residues 50-66 of SEQ ID NO:196 (CDR-H2 h1A11.H2); residues 99-107 of SEQ ID NO:196 (CDR-H3 h1A11.H2); residues 24-34 of SEQ ID NO:197 (CDR-L1 h1A11VL.1), residues 50-56 of SEQ ID NO:197 (CDR-L2 h1A11VL.1); residues 89-97 of SEQ ID NO:197 (CDR-L3 h1A11VL.1); residues 24-34 of SEQ ID NO:198 (CDR-L1 h1A11.A2), residues 50-56 of SEQ ID NO:198 (CDR-L2 h1A11.A2); residues 89-97 of SEQ ID NO:198 (CDR-L3 h1A11.A2); residues 24-34 of SEQ ID NO:199 (CDR-L1 h1A11.A12), residues 50-56 of SEQ ID NO:199 (CDR-L2 h1A11.A12); residues 89-97 of SEQ ID NO:199 (CDR-L3 h1A11.A12); residues 24-34 of SEQ ID NO:200 (CDR-L1 h1A11.A7), residues 50-56 of SEQ ID NO:200 (CDR-L2 h1A11.A7); residues 89-97 of SEQ ID NO:200 (CDR-L3 h1A11.A7); residues 24-34 of SEQ ID NO:201 (CDR-L1 h1A11.B4), residues 50-56 of SEQ ID NO:201 (CDR-L2 h1A11.B4); residues 89-97 of SEQ ID NO:201 (CDR-L3 h1A11.B4); residues 24-34 of SEQ ID NO:202 (CDR-L1 h1A11.B5), residues 50-56 of SEQ ID NO:202 (CDR-L2 h1A11.B5); residues 89-97 of SEQ ID NO:202 (CDR-L3 h1A11.B5); residues 24-34 of SEQ ID NO:203 (CDR-L1 h1A11.E12), residues 50-56 of SEQ ID NO:203 (CDR-L2 h1A11.E12); and residues 89-97 of SEQ ID NO:203 (CDR-L3 h1A11.E12).
4 . The binding protein according to claim 2 , wherein said binding protein comprises at least three CDRs.
5 . The binding protein according to claim 4 , wherein said at least three CDRs are selected from a variable domain CDR set selected from the group consisting of:
VH 38H12 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:157
CDR-H2: residues 50-66 of SEQ ID NO:157
CDR-H3 residues 99-107 of SEQ ID NO:157
VL 38H12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:158
CDR-L2: residues 50-56 of SEQ ID NO:158
CDR-L3: residues 89-97 of SEQ ID NO:158
VH 1A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:159
CDR-H2: residues 50-66 of SEQ ID NO:159
CDR-H3: residues 99-107 of SEQ ID NO:159
VL 1A11 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:160
CDR-L2: residues 50-56 of SEQ ID NO:160
CDR-L3: residues 89-97 of SEQ ID NO:160
VH 37D10 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:161
CDR-H2: residues 50-68 of SEQ ID NO:161
CDR-H3: residues 101-111 of SEQ ID NO:161
VL 37D10 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:162
CDR-L2: residues 50-56 of SEQ ID NO:162
CDR-L3: residues 89-97 of SEQ ID NO:162
VH 32C7 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:163
CDR-H2: residues 50-66 of SEQ ID NO:163
CDR-H3: residues 99-105 of SEQ ID NO:163
VL 32C7 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:164
CDR-L2: residues 50-56 of SEQ ID NO:164
CDR-L3: residues 89-98 of SEQ ID NO:164
VH 14G1 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:165
CDR-H2: residues 50-66 of SEQ ID NO:165
CDR-H3: residues 99-105 of SEQ ID NO:165
VL 14G1 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:166
CDR-L2: residues 50-56 of SEQ ID NO:166
CDR-L3: residues 89-97 of SEQ ID NO:166
VH 14A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:167
CDR-H2: residues 50-66 of SEQ ID NO:167
CDR-H3: residues 99-110 of SEQ ID NO:167
VL 14A11 CDR Set
CDR-L1: residues 23-37 of SEQ ID NO:168
CDR-L2: residues 53-59 of SEQ ID NO:168
CDR-L3: residues 92-100 of SEQ ID NO:168
VH 15D6 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:169
CDR-H2: residues 50-66 of SEQ ID NO:169
CDR-H3: residues 99-110 of SEQ ID NO:169
VL 15D6 CDR Set
CDR-L1: residues 23-37 of SEQ ID NO:170
CDR-L2: residues 53-59 of SEQ ID NO:170
CDR-L3: residues 92-100 of SEQ ID NO:170
VH VH.1 1A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:171
CDR-H2: residues 50-66 of SEQ ID NO:171
CDR-H3: residues 99-107 of SEQ ID NO:171
VH VH.1a 1A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:172
CDR-H2: residues 50-66 of SEQ ID NO:172
CDR-H3: residues 99-107 of SEQ ID NO:172
VH VH.1b 1A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:173
CDR-H2: residues 50-66 of SEQ ID NO:173
CDR-H3: residues 99-107 of SEQ ID NO:173
VH VH.2a 1A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:174
CDR-H2: residues 50-66 of SEQ ID NO:174
CDR-H3: residues 99-107 of SEQ ID NO:174
VL VL.1 1A11 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:175
CDR-L2: residues 50-56 of SEQ ID NO:175
CDR-L3: residues 89-97 of SEQ ID NO:175
VL VL.1a 1A11 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:176
CDR-L2: residues 50-56 of SEQ ID NO:176
CDR-L3: residues 89-97 of SEQ ID NO:176
VL VL.1b 1A11 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:177
CDR-L2: residues 50-56 of SEQ ID NO:177
CDR-L3: residues 89-97 of SEQ ID NO:177
VL VL.2a 1A11 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:178
CDR-L2: residues 50-56 of SEQ ID NO:178
CDR-L3: residues 89-97 of SEQ ID NO:178
VH VH.1 38H12 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:179
CDR-H2: residues 50-66 of SEQ ID NO:179
CDR-H3: residues 99-107 of SEQ ID NO:179
VH VH.1a 38H12 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:180
CDR-H2: residues 50-66 of SEQ ID NO:180
CDR-H3: residues 99-107 of SEQ ID NO:180
VH VH.1b 38H12 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:181
CDR-H2: residues 50-66 of SEQ ID NO:181
CDR-H3: residues 99-107 of SEQ ID NO:181
VH VH.2a 38H12 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:182
CDR-H2: residues 50-66 of SEQ ID NO:182
CDR-H3: residues 99-107 of SEQ ID NO:182
VL VL.1 38H12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:183
CDR-L2: residues 50-56 of SEQ ID NO:183
CDR-L3: residues 89-97 of SEQ ID NO:183
VL VL.1a 38H12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:184
CDR-L2: residues 50-56 of SEQ ID NO:184
CDR-L3: residues 89-97 of SEQ ID NO:184
VL VL.1b 38H12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:185
CDR-L2: residues 50-56 of SEQ ID NO:185
CDR-L3: residues 89-97 of SEQ ID NO:185
VL VL.2a 38H12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:186
CDR-L2: residues 50-56 of SEQ ID NO:186
CDR-L3: residues 89-97 of SEQ ID NO:186
VH hA11VH.1 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:187
CDR-H2: residues 50-66 of SEQ ID NO:187
CDR-H3: residues 99-107 of SEQ ID NO:187
VH hA11.A6 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:188
CDR-H2: residues 50-66 of SEQ ID NO:188
CDR-H3: residues 99-107 of SEQ ID NO:188
VH hA11.A8 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:189
CDR-H2: residues 50-66 of SEQ ID NO:189
CDR-H3: residues 99-107 of SEQ ID NO:189
VH hA11.C6 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:190
CDR-H2: residues 50-66 of SEQ ID NO:190
CDR-H3: residues 99-107 of SEQ ID NO:190
VH hA11.A11 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:191
CDR-H2: residues 50-66 of SEQ ID NO:191
CDR-H3: residues 99-107 of SEQ ID NO:191
VH hA11.B5 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:192
CDR-H2: residues 50-66 of SEQ ID NO:192
CDR-H3: residues 99-107 of SEQ ID NO:192
VH hA11.E12 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:193
CDR-H2: residues 50-66 of SEQ ID NO:193
CDR-H3: residues 99-107 of SEQ ID NO:193
VH hA11.G3 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:194
CDR-H2: residues 50-66 of SEQ ID NO:194
CDR-H3: residues 99-107 of SEQ ID NO:194
VH hA11.F5 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:195
CDR-H2: residues 50-66 of SEQ ID NO:195
CDR-H3: residues 99-107 of SEQ ID NO:195
VH hA11.H2 CDR Set
CDR-H1: residues 31-35 of SEQ ID NO:196
CDR-H2: residues 50-66 of SEQ ID NO:196
CDR-H3: residues 99-107 of SEQ ID NO:196
VL h1A11VL.1 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:197
CDR-L2: residues 50-56 of SEQ ID NO:197
CDR-L3: residues 89-97 of SEQ ID NO:197
VL h1A11.A2 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:198
CDR-L2: residues 50-56 of SEQ ID NO:198
CDR-L3: residues 89-97 of SEQ ID NO:198
VL h1A11.A12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:199
CDR-L2: residues 50-56 of SEQ ID NO:199
CDR-L3: residues 89-97 of SEQ ID NO:199
VL h1A11.A7 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:200
CDR-L2: residues 50-56 of SEQ ID NO:200
CDR-L3: residues 89-97 of SEQ ID NO:200
VL h1A11.B4 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:201
CDR-L2: residues 50-56 of SEQ ID NO:201
CDR-L3: residues 89-97 of SEQ ID NO:201
VL h1A11.B5 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:202
CDR-L2: residues 50-56 of SEQ ID NO:202
CDR-L3: residues 89-97 of SEQ ID NO:202
and
VL h1A11.E12 CDR Set
CDR-L1: residues 24-34 of SEQ ID NO:203
CDR-L2: residues 50-56 of SEQ ID NO:203
CDR-L3: residues 89-97 of SEQ ID NO: 203
6 . The binding protein according to claim 5 , comprising CDRs from at least two variable domain CDR sets.
7 . The binding protein according to claim 6 , wherein said at least two variable domain CDR sets are a pair of CDR sets selected from the group consisting of:
VH 38H12 CDR Set and VL 38H12 CDR Set,
VH 1A11 CDR Set and VL 1A11 CDR Set,
VH 37D10 CDR Set and VL 37D10 CDR Set,
VH 32C7 CDR Set and VL 32C7 CDR Set,
VH 14G1 Set and VL 14G1 CDR Set,
VH 14A11 CDR Set and VL 14A11 CDR Set,
VH 15D6 CDR Set and VL 15D6 CDR Set,
VH VH.1 1A11 CDR Set and VL VL.1 1A11 CDR Set,
VH VH.1 1A11 CDR Set and VL VL.1a 1A11 CDR Set,
VH VH.1 1A11 CDR Set and VL VL.1b 1A11 CDR Set,
VH VH.1 1A11 CDR Set and VL VL.2a 1A11 CDR Set,
VH VH.1a 1A11 CDR Set and VL VL.1 1A11 CDR Set,
VH VH.1a 1A11 CDR Set and VL VL.1a 1A11 CDR Set,
VH VH.1a 1A11 CDR Set and VL VL.1b 1A11 CDR Set,
VH VH.1a 1A11 CDR Set and VL VL.2a 1A11 CDR Set,
VH VH.1b 1A11 CDR Set and VL VL.1 1A11 CDR Set,
VH VH.1b 1A11 CDR Set and VL VL.1a 1A11 CDR Set,
VH VH.1b 1A11 CDR Set and VL VL.1b 1A11 CDR Set,
VH VH.1b 1A11 CDR Set and VL VL.2a 1A11 CDR Set,
VH VH.2a 1A11 CDR Set and VL VL.1 1A11 CDR Set,
VH VH.2a 1A11 CDR Set and VL VL.1a 1A11 CDR Set,
VH VH.2a 1A11 CDR Set and VL VL.1b 1A11 CDR Set,
VH VH.2a 1A11 CDR Set and VL VL.2a 1A11 CDR Set,
VH VH.1 38H12 CDR Set and VL VL.1 38H12 CDR Set,
VH VH.1 38H12 CDR Set and VL VL.1a 38H12 CDR Set,
VH VH.1 38H12 CDR Set and VL VL.1b 38H12 CDR Set,
VH VH.1 38H12 CDR Set and VL VL.2a 38H12 CDR Set,
VH VH.1a 38H12 CDR Set and VL VL.1 38H12 CDR Set,
VH VH.1a 38H12 CDR Set and VL VL.1a 38H12 CDR Set,
VH VH.1a 38H12 CDR Set and VL VL.1b 38H12 CDR Set,
VH VH.1a 38H12 CDR Set and VL VL.2a 38H12 CDR Set,
VH VH.1b 38H12 CDR Set and VL VL.1 38H12 CDR Set,
VH VH.1b 38H12 CDR Set and VL VL.1a 38H12 CDR Set,
VH VH.1b 38H12 CDR Set and VL VL.1b 38H12 CDR Set,
VH VH.1b 38H12 CDR Set and VL VL.2a 38H12 CDR Set,
VH VH.2a 38H12 CDR Set and VL VL.1 38H12 CDR Set,
VH VH.2a 38H12 CDR Set and VL VL.1a 38H12 CDR Set,
VH VH.2a 38H12 CDR Set and VL VL.1b 38H12 CDR Set,
VH VH.2a 38H12 CDR Set and VL VL.2a 38H12 CDR Set,
VH h1A11.A6 CDR Set and VL h1A11VL.1 CDR Set,
VH h1A11.C6 CDR Set and VL h1A11VL.1 CDR Set,
VH h1A11.A11 CDR Set and VL h1A11VL.1 CDR Set,
VH h1A11.A8 CDR Set and VL h1A11VL.1 CDR Set,
VH h1A11VH.1 CDR Set and VL h1A11.B4 CDR Set,
VH h1A11VH.1 CDR Set and VL h1A11.A7 CDR Set,
VH h1A11VH.1 CDR Set and VL h1A11.A12 CDR Set,
VH h1A11VH.1 CDR Set and VL h1A11.A2 CDR Set,
VH h1A11.B5 CDR Set and VL h1A11.B5 CDR Set,
VH h1A11.E12 CDR Set and VL h1A11.E12 CDR Set,
VH h1A11.G3 CDR Set and VL h1A11.E12 CDR Set,
VH h1A11.F5 CDR Set and VL h1A11.E12 CDR Set, and
VH h1A11.H2 CDR Set and VL h1A11.E12 CDR Set.
8 . The binding protein according to claim 4 , further comprising a human acceptor framework sequence.
9 . The binding protein according to claim 5 , further comprising a human acceptor framework sequence.
10 . The binding protein according to claim 6 , further comprising a human acceptor framework sequence.
11 . The binding protein according to claim 7 , further comprising a human acceptor framework sequence.
12 . The binding protein according to claim 8 , wherein said human acceptor framework sequence is selected from the group of acceptor sequences listed in Tables 3 and 4.
13 . The binding protein according to claim 9 , wherein said human acceptor framework sequence is selected from the group of acceptor sequences listed in Tables 3 and 4.
14 . The binding protein according to claim 10 , wherein said human acceptor framework sequence is selected from the group of acceptor sequences listed in Tables 3 and 4
15 . The binding protein according to claim 11 , wherein said human acceptor framework sequence is selected from the group of acceptor sequences listed in Tables 3 and 4.
16 . The binding protein according to claim 8 , wherein said human acceptor framework sequence is selected from any framework sequence in a variable region sequence selected from the group consisting of:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
171
180
188
196
VH.1 1A11
VH.1a 38H12
VH h1A11.A6
VH h1A11.H2
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
172
181
189
197
VH.1a 1A11
VH.1b 38H12
VH h1A11.A8
VL h1A11VL.1
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
173
182
190
198
VH.1b 1A11
VH.2a 38H12
VH h1A11.C6
VL h1A11.A2
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
174
183
191
199
VH.2a 1A11
VL.1 38H12
VH h1A11.A11
VL h1A11.A12
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
175
184
192
200
VL.1 1A11
VL.1a
VH h1A11.B5
VL h1A11.A7
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
176
185
193
201
VL.1a 1A11
VL.1b
VH h1A11.E12
VL h1A11.B4
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
177
186
194
202
VL.1b 1A11
VL.2a
VH h1A11.G3
VL h1A11.B5
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
178
187
195
203
VL.2a 1A11
VH h1A11VH.1
VH h1A11.F5
VL h1A11.E12
SEQ ID NO:
179 VH.1
38H12
17 . The binding protein according to claim 9 , wherein said human acceptor framework sequence is selected from any framework sequence in a variable region sequence selected from the group consisting of:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
171
180
188
196
VH.1 1A11
VH.1a 38H12
VH h1A11.A6
VH h1A11.H2
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
172
181
189
197
VH.1a 1A11
VH.1b 38H12
VH h1A11.A8
VL h1A11VL.1
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
173
182
190
198
VH.1b 1A11
VH.2a 38H12
VH h1A11.C6
VL h1A11.A2
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
174
183
191
199
VH.2a 1A11
VL.1 38H12
VH h1A11.A11
VL h1A11.A12
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
175
184
192
200
VL.1 1A11
VL.1a
VH h1A11.B5
VL h1A11.A7
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
176
185
193
201
VL.1a 1A11
VL.1b
VH h1A11.E12
VL h1A1 1.B4
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
177
186
194
202
VL.1b 1A11
VL.2a
VH h1A11.G3
VL h1A11.B5
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
178
187
195
203
VL.2a 1A11
VH h1A11VH.1
VH h1A11.F5
VL h1A11.E12
SEQ ID NO:
179
VH.1 38H12
18 . The binding protein according to claim 10 , wherein said human acceptor framework sequence is selected from any framework sequence in a variable region sequence selected from the group consisting of:
SEQ ID NO:
SEQ ID NO: 180
SEQ ID NO: 188
SEQ ID NO: 196
171
VH.1a 38H12
VH h1A11.A6
VH h1A11.H2
VH.1 1A11
SEQ ID NO:
SEQ ID NO: 181
SEQ ID NO: 189
SEQ ID NO: 197
172
VH.1b 38H12
VH h1A11.A8
VL h1A11VL.1
VH.1a 1A11
SEQ ID NO:
SEQ ID NO: 182
SEQ ID NO: 190
SEQ ID NO: 198
173
VH.2a 38H12
VH h1A11.C6
VL h1A11.A2
VH.1b 1A11
SEQ ID NO:
SEQ ID NO: 183
SEQ ID NO: 191
SEQ ID NO: 199
174
VL.1 38H12
VH h1A11.A11
VL h1A11.A12
VH.2a 1A11
SEQ ID NO:
SEQ ID NO: 184
SEQ ID NO: 192
SEQ ID NO: 200
175
VL.1a
VH h1A11.B5
VL h1A11.A7
VL.1 1A11
SEQ ID NO:
SEQ ID NO: 185
SEQ ID NO: 193
SEQ ID NO: 201
176
VL.1b
VH h1A11.E12
VL h1A11.B4
VL.1a 1A11
SEQ ID NO:
SEQ ID NO: 186
SEQ ID NO: 194
SEQ ID NO: 202
177
VL.2a
VH h1A11.G3
VL h1A11.B5
VL.1b 1A11
SEQ ID NO:
SEQ ID NO: 187
SEQ ID NO: 195
SEQ ID NO: 203
178
VH h1A11VH.1
VH h1A11.F5
VL h1A11.E12
VL.2a 1A11
SEQ ID NO:
179
VH.1 38H12
19 . The binding protein according to claim 11 , wherein said human acceptor framework sequence is selected from any framework sequence in a variable region sequence elected from the group consisting of:
SEQ ID NO:
SEQ ID NO: 180
SEQ ID NO: 188
SEQ ID NO: 196
171
VH.1a 38H12
VH h1A11.A6
VH h1A11.H2
VH.1 1A11
SEQ ID NO:
SEQ ID NO: 181
SEQ ID NO: 189
SEQ ID NO: 197
172
VH.1b 38H12
VH h1A11.A8
VL h1A11VL.1
VH.1a 1A11
SEQ ID NO:
SEQ ID NO: 182
SEQ ID NO: 190
SEQ ID NO: 198
173
VH.2a 38H12
VH h1A11.C6
VL h1A11.A2
VH.1b 1A11
SEQ ID NO:
SEQ ID NO: 183
SEQ ID NO: 191
SEQ ID NO: 199
174
VL.1 38H12
VH h1A11.A11
VL h1A11.A12
VH.2a 1A11
SEQ ID NO:
SEQ ID NO: 184
SEQ ID NO: 192
SEQ ID NO: 200
175
VL.1a
VH h1A11.B5
VL h1A11.A7
VL.1 1A11
SEQ ID NO:
SEQ ID NO: 185
SEQ ID NO: 193
SEQ ID NO: 201
176
VL.1b
VH h1A11.E12
VL h1A11.B4
VL.1a 1A11
SEQ ID NO:
SEQ ID NO: 186
SEQ ID NO: 194
SEQ ID NO: 202
177
VL.2a
VH h1A11.G3
VL h1A11.B5
VL.1b 1A11
SEQ ID NO:
SEQ ID NO: 187
SEQ ID NO: 195
SEQ ID NO: 203
178
VH h1A11VH.1
VH h1A11.F5
VL h1A11.E12
VL.2a 1A11
SEQ ID NO:
179
VH.1 38H12
20 . The binding protein according to claim 8 , wherein said binding protein comprises at least one acceptor framework sequence selected from the group consisting of:
heavy chain framework-1 (H-FR1): E-V-Q-L-V-E-S-G-G-G-L-V-Q-P-G-G-S-L-R-L-S-C-A-A-S-G-F-T-F-X 30 (SEQ ID NO:143), wherein X 30 is S, R, or G;
heavy chain framework-2 (H-FR2): W-V-R-Q-A-P-G-K-G-L-E-W-V-A (SEQ ID NO:144);
heavy chain framework-3 (H-FR3): R-F-T-I-S-R-D-N-A-K-X 11 -S-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-X 31 -R (SEQ ID NO:145), wherein;
X 11 is N or S; and
X 31 is A or S;
heavy chain framework-4 (H-FR4): W-G-Q-G-T-L-V-T-V-S-S(SEQ ID NO:146);
light chain framework-1 (L-FR1): D-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C(SEQ ID NO:147);
light chain framework-2 (L-FR2): W-Y-Q-Q-K-P-G-K-X 9 -P-K-L-L-I-X 15 (SEQ ID NO:148), wherein;
X 9 is A or S; and
X 15 is F or Y;
light chain framework-3 (L-FR3): G-V-P-S-R-F-S-G-S-G-S-G-T-D-X 15 -T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C (SEQ ID NO:149), wherein;
X 15 is F or S; and
light chain framework-4 (L-FR4): F-G-Q-G-T-K-L-E-I-K (SEQ ID NO:150).
21 . The binding protein according to claim 9 , wherein said binding protein comprises at least one acceptor framework sequence selected from the group consisting of:
heavy chain framework-1 (H-FR1): E-V-Q-L-V-E-S-G-G-G-L-V-Q-P-G-G-S-L-R-L-S-C-A-A-S-G-F-T-F-X 30 (SEQ ID NO:143), wherein X 30 is S, R, or G;
heavy chain framework-2 (H-FR2): W-V-R-Q-A-P-G-K-G-L-E-W-V-A (SEQ ID NO:144);
heavy chain framework-3 (H-FR3): R-F-T-I-S-R-D-N-A-K-X 11 -S-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-X 31 -R (SEQ ID NO:145), wherein;
X 11 is N or S; and
X 31 is A or S;
heavy chain framework-4 (H-FR4): W-G-Q-G-T-L-V-T-V-S-S(SEQ ID NO:146);
light chain framework-1 (L-FR1): D-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C(SEQ ID NO:147);
light chain framework-2 (L-FR2): (SEQ ID NO:148), wherein;
X 9 is A or S; and
X 15 is F or Y;
light chain framework-3 (L-FR3):
G-V-P-S-R-F-S-G-S-G-S-G-T-D-X 15 -T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C (SEQ ID NO:149), wherein;
X 15 is F or S; and
light chain framework-4 (L-FR4): F-G-Q-G-T-K-L-E-I-K (SEQ ID NO:150).
22 . The binding protein according to claim 10 , wherein said binding protein comprises at least one acceptor framework sequence selected from the group consisting of:
heavy chain framework-1 (H-FR1): E-V-Q-L-V-E-S-G-G-G-L-V-Q-P-G-G-S-L-R-L-S-C-A-A-S-G-F-T-F-X 30 (SEQ ID NO:143), wherein X 30 is S, R, or G;
heavy chain framework-2 (H-FR2): W-V-R-Q-A-P-G-K-G-L-E-W-V-A (SEQ ID NO:144);
heavy chain framework-3 (H-FR3): R-F-T-I-S-R-D-N-A-K-X 11 -S-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-X 31 -R (SEQ ID NO:145), wherein;
X 11 is N or S; and
X 31 is A or S;
heavy chain framework-4 (H-FR4): W-G-Q-G-T-L-V-T-V-S-S(SEQ ID NO:146);
light chain framework-1 (L-FR1): D-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C(SEQ ID NO:147);
light chain framework-2 (L-FR2): (SEQ ID NO:148), wherein;
X 9 is A or S; and
X 15 is F or Y;
light chain framework-3 (L-FR3): G-V-P-S-R-F-S-G-S-G-S-G-T-D-X 15 -T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C (SEQ ID NO:149), wherein;
X 15 is F or S; and
light chain framework-4 (L-FR4): F-G-Q-G-T-K-L-E-I-K (SEQ ID NO:150).
23 . The binding protein according to claim 11 , wherein said binding protein comprises at least one acceptor framework sequence selected from the group consisting of:
heavy chain framework-1 (H-FR1): E-V-Q-L-V-E-S-G-G-G-L-V-Q-P-G-G-S-L-R-L-S-C-A-A-S-G-F-T-F-X 30 (SEQ ID NO:143), wherein X 30 is S, R, or G;
heavy chain framework-2 (H-FR2): W-V-R-Q-A-P-G-K-G-L-E-W-V-A (SEQ ID NO:144);
heavy chain framework-3 (H-FR3): R-F-T-I-S-R-D-N-A-K-X 11 -S-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-X 31 -R (SEQ ID NO:145), wherein;
X 11 is N or S; and
X 31 is A or S;
heavy chain framework-4 (H-FR4): W-G-Q-G-T-L-V-T-V-S-S(SEQ ID NO:146);
light chain framework-1 (L-FR1): D-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C(SEQ ID NO:147);
light chain framework-2 (L-FR2): W-Y-Q-Q-K-P-G-K-X 9 -P-K-L-L-I-X 15 (SEQ ID NO:148), wherein;
X 9 is A or S; and
X 15 is F or Y;
light chain framework-3 (L-FR3): G-V-P-S-R-F-S-G-S-G-S-G-T-D-X 15 -T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C(SEQ ID NO:149), wherein;
X 15 is F or S; and
light chain framework-4 (L-FR4): F-G-Q-G-T-K-L-E-I-K (SEQ ID NO:150).
24 . The binding protein according to claim 2 , wherein said binding protein comprises a human acceptor framework sequence and at least one variable domain having an amino acid sequence selected from the group consisting of:
SEQ ID NO: 171
SEQ ID NO: 188
VH VH.1 1A11
VH h1A11.A6
SEQ ID NO: 172
SEQ ID NO: 189
VH VH.1a 1A11
VH h1A11.A8
SEQ ID NO: 173
SEQ ID NO: 190
VH VH.1b 1A11
VH h1A11.C6
SEQ ID NO: 174
SEQ ID NO: 191
VH VH.2a 1A11
VH h1A11.A11
SEQ ID NO: 175
SEQ ID NO: 192
VL VL.1 1A11
VH h1A11.B5
SEQ ID NO: 176
SEQ ID NO: 193
VL VL.1a 1A11
VH h1A11.E12
SEQ ID NO: 177
SEQ ID NO: 194
VL VL.1b
VH h1A11.G3
SEQ ID NO: 178
SEQ ID NO: 195
VL VL.2a 1A11
VH h1A11.F5
SEQ ID NO: 179
SEQ ID NO: 196
VH VH.1 38H12
VH h1A11.H2
SEQ ID NO: 180
SEQ ID NO: 197
VH VH.1a 38H12
VL h1A11VL.1
SEQ ID NO: 181
SEQ ID NO: 198
VH VH.1b 38H12
VL h1A11.A2
SEQ ID NO: 182
SEQ ID NO: 199
VH VH.2a 38H12
VL h1A11.A12
SEQ ID NO: 183
SEQ ID NO: 200
VL VL.1 38H12
VL h1A11.A7
SEQ ID NO: 184
SEQ ID NO: 201
VL VL.1a 38H12
VL h1A11.B4
SEQ ID NO: 185
SEQ ID NO: 202
VL VL.1b 38H12
VL h1A11.B5
SEQ ID NO: 186
SEQ ID NO: 203
VL VL.2a 38H12
VL h1A11.E12
SEQ ID NO: 187
VH h1A11VH.1
25 . The binding protein according to claim 24 , 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 NO: 171 and SEQ ID NO: 175
SEQ ID NO: 180 and SEQ ID
VH.1 and VL.1 1A11 (Table 12)
NO: 186
VH.1a and VL.2a (Table 16)
SEQ ID NO: 171 and SEQ ID NO: 176
SEQ ID NO: 181 and SEQ ID
VH.1 and VL.1a 1A11 (Table 12)
NO: 183
VH.1b and VL.1 (Table 16)
SEQ ID NO: 171 and SEQ ID NO: 177
SEQ ID NO: 181 and SEQ ID
VH.1 and VL.1b 1A11 (Table 12)
NO: 184
VH.1b and VL.1a (Table 16)
SEQ ID NO: 171 and SEQ ID NO: 178
SEQ ID NO: 181 and SEQ ID
VH.1 and VL.2a 1A11(Table 12)
NO: 185
VH.1b and VL.1b (Table 16)
SEQ ID NO: 172 and SEQ ID NO: 175
SEQ ID NO: 181 and SEQ ID
VH.1a and VL.1 1A11 (Table 12)
NO: 186
VH.1b and VL.2a (Table 16)
SEQ ID NO: 172 and SEQ ID NO: 176
SEQ ID NO: 182 and SEQ ID
VH.1a and VL.1a 1A11 (Table 12)
NO: 183
VH.2a and VL.1 (Table 16)
SEQ ID NO: 172 and SEQ ID NO: 177
SEQ ID NO: 182 and SEQ ID
VH.1a and VL.1b 1A11 (Table 12)
NO: 184
VH.2a and VL.1a (Table 16)
SEQ ID NO: 172 and SEQ ID NO: 178
SEQ ID NO: 182 and SEQ ID
VH.1a and VL.2a 1A11 (Table 12)
NO: 185
VH.2a and VL.1b (Table 16)
SEQ ID NO: 173 and SEQ ID NO: 175
SEQ ID NO: 182 and SEQ ID
VH.1b and VL.1 1A11 (Table 12)
NO: 186
VH.2a and VL.2a (Table 16)
SEQ ID NO: 173 and SEQ ID NO: 176
SEQ ID NO: 188 and SEQ ID
VH.1b and VL.1a 1A11 (Table 12)
NO: 197
h1A11.A6 VH and h1A11VL.1
Tables 20/21
SEQ ID NO: 173 and SEQ ID NO: 177
SEQ ID NO: 190 and SEQ ID
VH.1b and VL.1b 1A11 (Table 12)
NO: 197
h1A11.C6 VH and h1A11VL.1
Tables 20/21
SEQ ID NO: 173 and SEQ ID NO: 178
SEQ ID NO: 191 and SEQ ID
VH.1b and VL.2a 1A11 (Table 12)
NO: 197
h1A11.All VH and h1A11VL.1
Tables 20/21
SEQ ID NO: 174 and SEQ ID NO: 175
SEQ ID NO: 189 and SEQ ID
VH.2a and VL.1 1A11 (Table 12)
NO: 197
h1A11.A8 VH and h1A11 VL.1
Tables 20/21
SEQ ID NO: 174 and SEQ ID NO: 176
SEQ ID NO: 187 and SEQ ID
VH.2a and VL.1a 1A11 (Table 12)
NO: 201
h1A11VH.1 and h1A11.B4 VL
Tables 20/21
SEQ ID NO: 174 and SEQ ID NO: 177
SEQ ID NO: 187 and SEQ ID
VH.2a and VL.1b 1A11 (Table 12)
NO: 200
h1A11VH.1 and h1All.A7 VL
Tables 20/21
SEQ ID NO: 174 and SEQ ID NO: 178
SEQ ID NO: 187 and SEQ ID
VH.2a and VL.2a 1A11 (Table 12)
NO: 199
h1A11VH.1 and h1All.A12 VL
Tables 20/21
SEQ ID NO: 179 and SEQ ID NO: 183
SEQ ID NO: 187 and SEQ ID
VH.1 and VL.1 38H12 (Table 16)
NO: 198
h1A11VH.1 VH and
h1All.A2 VL
Tables 20/21
SEQ ID NO: 179 and SEQ ID NO: 184
SEQ ID NO: 192 and SEQ ID
VH.1 and VL.1a 38H12 (Table 16)
NO: 202
h1A11.B5 VH and h1All.B5 VL
Tables 20/21
SEQ ID NO: 179 and SEQ ID NO: 185
SEQ ID NO: 193 and SEQ ID
VH.1 and VL.1b 38H12 (Table 16)
NO: 203
h1A11.E12 VH and
h1All.E12 VL
Tables 20/21
SEQ ID NO: 179 and SEQ ID NO: 186
SEQ ID NO: 194 and SEQ ID
VH.1 and VL.2a 38H12 (Table 16)
NO: 203
h1A11.G3 VH and
h1All.E12 VL
Tables 20/21
SEQ ID NO: 180 and SEQ ID NO: 183
SEQ ID NO: 195 and SEQ ID
VH.1a and VL.1 38H12 (Table 16)
NO: 203
h1A11.F5 VH and
h1All.E12 VL
Tables 20/21
SEQ ID NO: 180 and SEQ ID NO: 184
SEQ ID NO: 196 and SEQ ID
VH.1a and VL.1a 38H12 (Table 16)
NO: 203
h1A1 1.H2 VH and
h1All.E12 VL
Tables 20/21
SEQ ID NO: 180 and SEQ ID NO: 185
VH.1a and VL.1b 38H12 (Table 16)
26 . The binding protein according to claim 8 , wherein said human acceptor framework sequence 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 residue capable of interacting with a CDR;
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.
27 . The binding protein according to claim 11 , wherein said human acceptor framework sequence 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 residue capable of interacting with a CDR;
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 DR1 and a Kabat-defined first heavy chain framework.
28 . The binding protein according to claim 25 , wherein said human acceptor framework sequence 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 residue capable of interacting with a CDR;
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.
29 . The binding protein according to claim 26 , wherein the binding protein comprises a consensus human variable domain sequence.
30 . The binding protein according to claim 27 , wherein the binding protein comprises a consensus human variable domain sequence.
31 . The binding protein according to claim 28 , wherein the binding protein comprises a consensus human variable domain sequence.
32 . The binding protein according to 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 a sequence of a human germline acceptor framework and comprises at least 70 amino acid residues identical to the human germline acceptor framework.
33 . The binding protein according to claim 11 , 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 a sequence of a human germline acceptor framework and comprises at least 70 amino acid residues identical to the human germline acceptor framework.
34 . The binding protein according to claim 25 , 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 a sequence of a human germline acceptor framework and comprises at least 70 amino acid residues identical to the human germline acceptor framework.
35 . The binding protein according to claim 2 , wherein said binding protein comprises at least one variable domain having an amino acid sequence selected from the group consisting of:
SEQ ID NO: 157
SEQ ID NO: 181
VH 38H12
VH VH.1b 38H12
SEQ ID NO: 158
SEQ ID NO: 182
VL 38H12
VH VH.2a 38H12
SEQ ID NO: 159
SEQ ID NO: 182
VH 1A11
VL VL.1 38H12
SEQ ID NO: 160
SEQ ID NO: 184
VL 1A11
VL VL.1a 38H12
SEQ ID NO: 161
SEQ ID NO: 185
VH 37D10
VL VL.1b 38H12
SEQ ID NO: 162
SEQ ID NO: 186
VL 37D10
VL VL.2a 38H12
SEQ ID NO: 163
SEQ ID NO: 187
VH 32C7
VH h1A11VH.1
SEQ ID NO: 164
SEQ ID NO: 188
VL 32C7
VH h1A11.A6
SEQ ID NO: 165
SEQ ID NO: 189
VH 14G1
VH h1A11.A8
SEQ ID NO: 166
SEQ ID NO: 190
VL 14G1
VH h1A11.C6
SEQ ID NO: 167
SEQ ID NO: 191
VH 14A11
VH h1A11.A11
SEQ ID NO: 168
SEQ ID NO: 192
VL 14A11
VH h1A11.B5
SEQ ID NO: 169
SEQ ID NO: 193
VH 15D6
VH h1A11.E12
SEQ ID NO: 170
SEQ ID NO: 194
VL 15D6
VH h1A11.G3
SEQ ID NO: 171
SEQ ID NO: 195
VH VH.1 1A11
VH h1A11.F5
SEQ ID NO: 172
SEQ ID NO: 196
VH VH.1a 1A11
VH h1A11.H2
SEQ ID NO: 173
SEQ ID NO: 197
VH VH.1b 1A11
VL h1A11VL.1
SEQ ID NO: 174
SEQ ID NO: 198
VH VH.2a 1A11
VL h1A11.A2
SEQ ID NO: 175
SEQ ID NO: 199
VL VL.1 1A11
VL h1A11.A12
SEQ ID NO: 176
SEQ ID NO: 200
VL VL.1a 1A11
VL h1A11.A7
SEQ ID NO: 177
SEQ ID NO: 201
VL VL.1b
VL h1A11.B4
SEQ ID NO: 178
SEQ ID NO: 202
VL VL.2a 1A11
VL h1A11.B5
SEQ ID NO: 179
SEQ ID NO: 203
VH VH.1 38H12
VL h1A11.E12
SEQ ID NO: 180
VH VH.1a 38H12
36 . The binding protein according to claim 35 , 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 NO: 157 and SEQ ID NO: 158
SEQ ID NO: 179 and SEQ ID
38H12
NO: 186
VH.1 and VL.2a 38H12
SEQ ID NO: 159 and SEQ ID NO: 160
SEQ ID NO: 180 and SEQ ID
1A11
NO: 183
VH.1a and VL.1 38H12
SEQ ID NO: 161 and SEQ ID NO: 162
SEQ ID NO: 180 and SEQ ID
37D10
NO: 184
VH.1a and VL.1a 38H12
SEQ ID NO: 163 and SEQ ID NO: 164
SEQ ID NO: 180 and SEQ ID
32C7
NO: 185
VH.1a and VL.1b 38H12
SEQ ID NO: 165 and SEQ ID NO: 166
SEQ ID NO: 180 and SEQ ID
14G1
NO: 186
VH.1a and VL.2a 38H12
SEQ ID NO: 167 and SEQ ID NO: 168
SEQ ID NO: 181 and SEQ ID
14A11
NO: 183
VH.1b and VL.1 38H12
SEQ ID NO: 169 and SEQ ID NO: 170
SEQ ID NO: 181 and SEQ ID
15D6
NO: 184
VH.1b and VL.1a 38H12
SEQ ID NO: 171 and SEQ ID NO: 175
SEQ ID NO: 181 and SEQ ID
VH.1 and VL.1 1A11 (Table 11)
NO: 185
VH.1b and VL.1b 38H12
SEQ ID NO: 171 and SEQ ID NO: 176
SEQ ID NO: 181 and SEQ ID
VH.1 and VL.1a 1A11
NO: 186
VH.1b and VL.2a 38H12
SEQ ID NO: 171 and SEQ ID NO: 177
SEQ ID NO: 182 and SEQ ID
VH.1 and VL.1b 1A11
NO: 183
VH.2a and VL.1 38H12
SEQ ID NO: 171 and SEQ ID NO: 178
SEQ ID NO: 182 and SEQ ID
VH.1 and VL.2a
NO: 184
VH.2a and VL.1a 38H12
SEQ ID NO: 172 and SEQ ID NO: 175
SEQ ID NO: 182 and SEQ ID
VH.1a and VL.1
NO: 185
VH.2a and VL.1b 38H12
SEQ ID NO: 172 and SEQ ID NO: 176
SEQ ID NO: 182 and SEQ ID
VH.1a and VL.1a
NO: 186
VH.2a and VL.2a 38H12
SEQ ID NO: 172 and SEQ ID NO: 177
SEQ ID NO: 188 and SEQ ID
VH.1a and VL.1b
NO: 197 h1A11.A6
VH and h1A11VL.1
SEQ ID NO: 172 and SEQ ID NO: 178
SEQ ID NO: 190 and SEQ ID
VH.1a and VL.2a
NO: 197 h1A11.C6
VH and h1A11VL.1
SEQ ID NO: 173 and SEQ ID NO: 175
SEQ ID NO: 191 and SEQ ID
VH.1b and VL.1
NO: 197 h1A11.All
VH and h1A11VL.1
SEQ ID NO: 173 and SEQ ID NO: 176
SEQ ID NO: 189 and SEQ ID
VH.1b and VL.1a
NO: 197 h1A11.A8
VH and h1A11 VL.1
SEQ ID NO: 173 and SEQ ID NO: 177
SEQ ID NO: 187 and SEQ ID
VH.1b and VL.1b
NO: 201 h1A11VH.1
and h1A11.B4 VL
SEQ ID NO: 173 and SEQ ID NO: 178
SEQ ID NO: 187 and SEQ ID
VH.1b and VL.2a
NO: 200
h1A11VH.1 and h1All.A7 VL
SEQ ID NO: 174 and SEQ ID NO: 175
SEQ ID NO: 187 and SEQ ID
VH.2a and VL.1
NO: 199 h1A11VH.1
and h1All.A12 VL
SEQ ID NO: 174 and SEQ ID NO: 176
SEQ ID NO: 187 and SEQ ID
VH.2a and VL.1a
NO: 198 h1A11VH.1
VH and h1All.A2 VL
SEQ ID NO: 174 and SEQ ID NO: 177
SEQ ID NO: 192 and SEQ ID
VH.2a and VL.1b
NO: 202 h1A11.B5
VH and h1All.B5 VL
SEQ ID NO: 174 and SEQ ID NO: 178
SEQ ID NO: 193 and SEQ ID
VH.2a and VL.2a
NO: 203 h1A11.E12
VH and h1All.E12 VL
SEQ ID NO: 179 and SEQ ID NO: 183
SEQ ID NO: 194 and SEQ ID
VH.1 and VL.1 38H12
NO: 203 h1A11.G3
VH and h1All.E12 VL
SEQ ID NO: 179 and SEQ ID NO: 184
SEQ ID NO: 195 and SEQ ID
VH.1 and VL.1a 38H12
NO: 203 h1A11.F5 VH
and h1All.E12 VL
SEQ ID NO: 179 and SEQ ID NO: 185
SEQ ID NO: 196 and SEQ ID
VH.1 and VL.1b 38H12
NO: 203 h1A11.H2
VH and h1All.E12 VL
37 . The DLL4 binding protein according to claim 2 , wherein said binding protein is capable of blocking DLL4 interaction with a Notch protein.
38 . The DLL4 binding protein according to claim 36 , 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.
39 . The DLL4 binding protein according to claim 2 , wherein said binding protein is capable of modulating a biological function of DLL4.
40 . The DLL4 binding protein according to claim 2 , wherein said binding protein is capable of neutralizing a biological function of DLL4.
41 . The DLL4 binding protein according to claim 2 , wherein said binding protein is capable of inhibiting VEGFR2 activity, VEGFR1 activity, or both.
42 . The DLL4 binding protein according to claim 2 , wherein said binding protein is capable of diminishing the ability of DLL4 to bind to its receptor.
43 . The DLL4 binding protein according to claim 2 , wherein said binding protein is capable of inhibiting normal angiogenesis.
44 . The DLL4 binding protein according to claim 2 , wherein said binding protein has an on rate constant (Kon) to DLL4 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 , as measured by surface plasmon resonance.
45 . The DLL4 binding protein according to claim 8 , wherein said binding protein has an on rate constant (Kon) to DLL4 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 , as measured by surface plasmon resonance.
46 . The DLL4 binding protein according to claim 24 , wherein said binding protein has an on rate constant (Kon) to DLL4 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 , as measured by surface plasmon resonance.
47 . The DLL4 binding protein according to claim 2 , wherein said binding protein has an off rate constant (Koff) to DLL4 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 , as measured by surface plasmon resonance.
48 . The DLL4 binding protein according to claim 8 , wherein said binding protein has an off rate constant (Koff) to DLL4 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 , as measured by surface plasmon resonance.
49 . The DLL4 binding protein according to claim 24 , wherein said binding protein has an off rate constant (Koff) to DLL4 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 , as measured by surface plasmon resonance.
50 . The DLL4 binding protein according to claim 2 , wherein said binding protein has a dissociation constant (K D ) to DLL4 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.
51 . The DLL4 binding protein according to claim 8 , wherein said binding protein has a dissociation constant (K D ) to DLL4 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.
52 . The DLL4 binding protein according to claim 24 , wherein said binding protein has a dissociation constant (K D ) to DLL4 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.
53 . An antibody construct comprising a binding protein described in claim 2 , said antibody construct further comprising a linker polypeptide or an immunoglobulin constant domain.
54 . The antibody construct according to claim 53 , selected from the group consisting of:
an immunoglobulin molecule,
a monoclonal antibody,
a chimeric antibody,
a CDR-grafted antibody,
a Fab,
a Fab′,
a F(ab′) 2 ,
an Fv,
a disulfide linked Fv,
an scFv,
a single domain antibody,
a diabody,
a multispecific antibody,
a dual specific antibody, and
a bispecific antibody.
55 . The antibody construct according to claim 53 , 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,
and
a human IgA constant domain.
56 . The antibody construct according to claim 53 , comprising an immunoglobulin constant domain having an amino acid sequence selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and combinations thereof.
57 . An antibody conjugate comprising an antibody construct as described in claim 53 , said antibody conjugate further comprising an agent selected from the group consisting of: an imaging agent, a therapeutic agent, a cytotoxic agent, and an immunoadhesion molecule.
58 . The antibody conjugate according to claim 57 , wherein said agent is an imaging agent selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
59 . The antibody conjugate according to claim 57 , wherein said imaging agent 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.
60 . The antibody conjugate according to claim 57 , wherein said agent is a therapeutic or cytotoxic agent 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, a toxin, and an apoptotic agent.
61 . The antibody construct according to claim 53 , wherein said binding protein possesses a human glycosylation pattern.
62 . The antibody conjugate according to claim 57 , wherein said binding protein possesses a human glycosylation pattern.
63 . The binding protein according to claim 4 , wherein said binding protein exists as a crystal.
64 . The antibody construct according to claim 53 , wherein said antibody construct exists as a crystal.
65 . The antibody conjugate according to claim 57 , wherein said antibody construct exists as a crystal.
66 . The binding protein according to claim 63 , wherein said crystal is a carrier-free pharmaceutical controlled release crystal.
67 . The antibody construct according to claim 64 , wherein said crystal is a carrier-free pharmaceutical controlled release crystal.
68 . The antibody conjugate according to claim 65 , wherein said crystal is a carrier-free pharmaceutical controlled release crystal.
69 . The binding protein according to claim 63 , wherein said binding protein crystal has a greater half life in vivo than the soluble counterpart of the binding protein.
70 . The antibody construct according to claim 64 , wherein said antibody construct crystal has a greater half life in vivo than the soluble counterpart of the antibody construct.
71 . The antibody conjugate according to claim 65 , wherein said antibody conjugate crystal has a greater half life in vivo than the soluble counterpart of the antibody conjugate.
72 . The binding protein according to claim 63 , wherein said binding protein crystal retains biological activity of the non-crystal form of the binding protein.
73 . The antibody construct according to claim 64 , wherein said antibody construct crystal retains biological activity of the non-crystal form of the antibody construct.
74 . The antibody conjugate according to claim 65 , wherein said antibody conjugate crystal retains biological activity of the non-crystal form of the antibody conjugate.
75 . An isolated nucleic acid encoding a binding protein amino acid sequence described in claim 2 .
76 . 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 a CDR-H1, a CDR-H2, and a CDR-H3 as described in claim 2 ; a polypeptide comprising a light chain variable domain, wherein the light chain variable domain comprises one or more of a CDR-L1, a CDR-L2, and a CDR-L3 as described in claim 2 ; and a combination of both polypeptides.
77 . A vector comprising the isolated nucleic acid according to claim 76 .
78 . The vector according to claim 77 , wherein the vector is selected from the group consisting of: pcDNA, pTT, pTT3, pEFBOS, pBV, pJV, and pBJ.
79 . A host cell comprising the vector described in claim 77 .
80 . The host cell according to claim 79 , wherein said host cell is a prokaryotic cell.
81 . The host cell according to claim 80 , wherein said host cell is Escherichia coli.
82 . The host cell according to claim 79 , wherein said host cell is a eukaryotic cell.
83 . The host cell according to claim 82 , wherein said eukaryotic cell is selected from the group consisting of: a protist cell, an animal cell, a plant cell, and a fungal cell.
84 . The host cell according to claim 83 , wherein said eukaryotic cell is an animal cell selected from the group consisting of: a mammalian cell, an avian cell, and an insect cell.
85 . The host cell according to claim 84 , wherein said mammalian cell is a CHO cell.
86 . The host cell according to claim 84 , wherein said mammalian cell is a COS cell.
87 . The host cell according to claim 83 , wherein said fungal cell is Saccharomyces cerevisiae.
88 . The host cell according to claim 84 , wherein said insect cell is an Sf9 cell.
89 . A method of producing a binding protein that binds human DLL4, comprising culturing the host cell as described in claim 79 in a culture medium under conditions sufficient to produce a binding protein that binds human DLL4.
90 . A DLL4 binding protein produced according to the method of claim 89 .
91 . A composition for the release of a binding protein said composition comprising:
(a) a formulation, wherein said formulation comprises a crystallized binding protein according as described in claim 63 , and an ingredient; and
(b) at least one polymeric carrier.
92 . The composition according to claim 93 , 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-hydroxybutyrate), 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 polysaccharides, blends and copolymers thereof.
93 . The composition according to claim 91 , wherein said ingredient is selected from the group consisting of albumin, sucrose, trehalose, lactitol, gelatin, hydroxypropyl-□-cyclodextrin, methoxypolyethylene glycol and polyethylene glycol.
94 . A method for treating a mammal comprising the step of administering to the mammal an effective amount of the composition as described in claim 91 .
95 . A pharmaceutical composition comprising a DLL4 binding protein as described in claim 2 , and a pharmaceutically acceptable carrier.
96 . The pharmaceutical composition of claim 95 , further comprising at least one additional agent for treating a disorder in which DLL4 activity is detrimental.
97 . The pharmaceutical composition of claim 95 , wherein said additional agent is selected from the group consisting of: a therapeutic agent; an imaging agent; an antineoplastic agent; a chemotherapeutic agent; an angiogenesis inhibitor; an anti-VEGF antibody; an anti-EGFR antibody; an anti-cMet antibody; an anti-ErbB3 antibody; an anti-HER2 antibody; an anti-CD20 antibody; aflibercept; a kinase inhibitor; a co-stimulation molecule blocker; an anti-B7.2 antibody; a CTLA4-Ig; an adhesion molecule blocker; an anti-E selectin antibody; an anti-L selectin antibody; an anti-cytokine antibody or functional fragment thereof; an anti-IL-18 antibody; an anti-TNF antibody; anti-IL-6 antibody; methotrexate; a corticosteroid; a cyclosporin; a rapamycin; FK506; a DNA alkylating agent; cisplatin; carboplatin; an anti-tubulin agent; paclitaxel; docetaxel; doxorubicin; gemcitabine; gemzar; an anthracycline; adriamycin; a topoisiomersase I inhibitor; a topoisomerase II inhibitor; 5-fluorouracil (5-FU); leucovorin; irinotecan; a receptor tyrosine kinase inhibitor, an apoptosis inhibitor; a Bcl2/Bclx inhibitor; erlotinib, gefitinib, a COX-2 inhibitor, celecoxib, cyclosporin; rapamycin; a detectable label or reporter molecule; a TNF antagonist; an antirheumatic; a muscle relaxant; a narcotic; an analgesic; an anesthetic; a sedative; a local anesthetic; a neuromuscular blocker; an antimicrobial agent; an antipsoriatic agent; a corticosteroid; an anabolic steroid; an erythropoietin; an immunization; an immunoglobulin; an immunosuppressive agent; a growth hormone; a hormone replacement drug; a radiopharmaceutical drug; an antidepressant; an antipsychotic drug; a stimulant; an asthma medication; a beta agonist; an inhaled steroid; an epinephrine; an epinephrine analog thereof; a cytokine; and a cytokine antagonist.
98 . A method for reducing human DLL4□ activity comprising contacting human DLL4 with the binding protein as described in claim 2 such that human DLL4 activity is reduced.
99 . 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 a binding protein as described in claim 2 such that human DLL4 activity in the human subject is reduced.
100 . A method for treating a subject for a disease or a disorder in which DLL4 activity is detrimental by administering to the subject a DLL4 binding protein as described in claim 2 such that treatment is achieved.
101 . The method of claim 100 , 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, hemangioma, melanoma, sarcoma, brain tumor, nerve cancer, eye tumor, meninges cancer, solid tumors from hematopoietic malignancy, tumor metastases, ocular neovascularization, edema, rheumatoid arthritis, atherosclerotic plaques, Crohn's disease, inflammatory bowel disease, refractory ascites, psoriasis, sarcoidosis, arterial arteriosclerosis, sepsis, peptic ulcers, burns, pancreatitis, polycystic ovarian disease (POD), endometriosis, uterine fibroid, benign prostate hypertrophy, T-cell acute lymphoblastic leukemia (T-ALL), cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), multiple sclerosis (MS), tetralogy of Fallot (TOF), Alagille syndrome (AS), macular degeneration and age-related macular degeneration diseases, and other angiogenesis independent and dependent diseases characterized by aberrant DLL4 activity.
102 . The method according to claim 101 , wherein the disorder is a primary and metastatic cancer.
103 . The method according to claim 101 , wherein the urinary tract cancer is selected from the group consisting of renal cancer, bladder cancer, and urothelium cancer.
104 . The method according to claim 101 , wherein the female genital tract cancer is selected from the group consisting of cervical cancer, uterine cancer, ovarian cancer, choriocarcinoma, and gestational trophoblastic disease.
105 . The method according to claim 101 , wherein the male genital tract cancer is selected from the group consisting of prostate cancer, seminal vesicles cancer, testicular cancer, and germ cell tumor.
106 . The method according to claim 101 , wherein the endocrine gland cancer is selected from the group consisting of thyroid cancer, adrenal cancer, and pituitary gland cancer.
107 . The method according to claim 101 , wherein the sarcoma is selected from the group consisting of a bone sarcoma, a soft tissue sarcoma, and Kaposi's sarcoma.
108 . The method according to claim 101 , 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 meningioma.
109 . The method according to claim 101 , 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.
110 . The method according to claim 101 , wherein the ocular neovascularization is selected from the group consisting of diabetic blindness, a retinopathy, an age-related macular degeneration, and a rubeosis.
111 . The method according to claim 101 , wherein said administering to the subject is by at least one mode selected from the group consisting of: parenteral, subcutaneous, intramuscular, intravenous, intraarterial, intraarticular, intrabronchial, intraabdominal, intracapsular, intracartilaginous, intracavitary, intracelial, intracerebellar, intracerebroventricular, intracolic, intracervical, intragastric, intrahepatic, intramyocardial, intraosteal, intrapelvic, intrapericardiac, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravesical, bolus, vaginal, rectal, buccal, sublingual, intranasal, and transdermal.
112 . A method of treating a patient suffering from a disorder in which DLL4 is detrimental comprising the step of administering a DLL4 binding protein as described in claim 2 before, concurrent with, or after the administration of a therapeutically effective amount 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; a corticosteroid; a cyclosporine; a rapamycin; FK506; a non-steroidal anti-inflammatory agent (NSAID); a radiotherapeutic agent; an antineoplastic agent; a chemotherapeutic agent; a DNA alkylating agent; cisplatin; carboplatin; an anti-tubulin agent; paclitaxel; docetaxel; taxol; doxorubicin; gemcitabine; gemzar; an anthracycline; adriamycin; a topoisomerase I inhibitor; a topoisomerase II inhibitor; 5-fluorouracil (5-FU); leucovorin; irinotecan; a receptor tyrosine kinase inhibitor; erlotinib; gefitinib; a COX-2 inhibitor; celecoxib; a kinase inhibitor; an angiogenesis inhibitor; an anti-VEGF antibody; aflibercept; a co-stimulation molecule blocker; an anti-B7.1 antibody; an anti-B7.2 antibody; a CTLA4-Ig; an anti-CD20 antibody; an adhesion molecule blocker; an anti-LFA-1 antibody; an anti-E selectin antibody; and anti-L selectin antibody; a small molecule inhibitor; an anti-cytokine antibody or functional fragment thereof; an anti-IL-18 antibody; anti-TNF antibody; an anti-IL-6 antibody; an anti-cytokine receptor antibody; a detectable label or reporter; a TNF antagonist; an antirheumatic; a muscle relaxant; a narcotic; an analgesic; an anesthetic; a sedative; a local anesthetic; a neuromuscular blocker; an antimicrobial agent; an antipsoriatic drug; a corticosteroid; an anabolic steroid; an erythropoietin; an immunization; an immunoglobulin; an immunosuppressive agent; a growth hormone; a hormone replacement drug; a radiopharmaceutical drug; an antidepressant; an antipsychotic drug; a stimulant; an asthma medication; a beta agonist; an inhaled steroid; an epinephrine; an epinephrine analog; a cytokine; and a cytokine antagonist.
113 . A binding protein according to claim 2 , wherein said binding protein is an antibody.
114 . The antibody according to claim 113 , wherein the antibody is selected from the group consisting of a monoclonal antibody, a full-length tetrameric immunoglobulin, an IgG molecule, an IgG 1 molecule, a chimeric antibody, a CDR-grafted antibody, a humanized antibody, and an affinity matured antibody.
115 . The antibody according to claim 114 , wherein the antibody is a monoclonal antibody.
116 . The binding protein according to claim 36 , wherein said two variable domains have the amino acid sequences selected from the group consisting of:
SEQ ID NO:188 and SEQ ID NO:197,
SEQ ID NO:190 and SEQ ID NO:197, and
SEQ ID NO:191 and SEQ ID NO:197.
117 . The binding protein according to claim 36 , wherein said two variable domains have the amino acid sequences of SEQ ID NO:181 and SEQ ID NO:185.