Tumor-specific recognition molecules
The invention relates to recognition molecules which are directed towards tumors and can be used in the diagnosis and therapy of tumor diseases.
1. A method of providing a molecule for tumor recognition comprising:
a. providing a recombinant recognition molecule which comprises a variable heavy chain and a variable light chain and which specifically binds to core 1 antigen, wherein
the variable heavy chain comprises (i) a CDRH1 region that comprises the amino acid sequence of SEQ ID NO: 1, or a variant thereof having at least 70% homology with SEQ ID NO:1; (ii) a CDRH2 region that comprises the amino acid sequence of any one of SEQ ID NO: 2-3, or a variant thereof having at least 70% homology with SEQ ID NO:2 or SEQ ID NO:3; and (iii) a CDRH3 region comprises the amino acid sequence of any one of SEQ ID NO: 4-6, or a variant thereof having at least 70% homology to any one of SEQ ID NO:4-6; and
the variable light chain comprises (i) a CDRL1 region that comprises the amino acid sequence of any one of SEQ ID NO: 7-9, or a variant thereof having at least 70% homology to any one of SEQ ID NOS:7-9; (ii) a CDRL2 region that comprises the amino acid sequence of any one of SEQ ID NO: 10-11, or a variant thereof having at least 70% homology to any one of SEQ ID NO:10-11; and a CDRL3 region that comprises the amino acid sequence of any one of SEQ ID NO: 12-13 or a variant thereof having at least 70% homology to any one of SEQ ID NO: 12-13,
b. modifying the recombinant recognition molecule, and
c. selecting a modified recombinant recognition molecule whose core 1 antigen specificity is retained.
2. The method of claim 1 , wherein the modification is a mutation, deletion or insertion.
3. The method of claim 1 , wherein the modification is a replacement by an equivalent canonical structure.
4. The method of claim 1 , wherein the CDRH1 region comprises the amino acid sequence of SEQ ID NO: 1.
5. The method of claim 1 , wherein the CDRH1 region variant has I, M, W, or V substituted for L at position 4 of SEQ ID NO:1.
6. The method of claim 1 , wherein the CDRH2 region comprises the amino acid sequence of any one of SEQ ID NO: 2-3.
7. The method of claim 1 , wherein the CDRH2 region variant is selected from (a) a variant that has T or A substituted for P at position 4 of SEQ ID NO: 2 or 3; (b) a variant that has D or N substituted for G or S at position 7 of SEQ ID NO:2 or 3; and (c) a variant that has T or A substituted for P at position 4 and D or N substituted for G or S at position 7 of SEQ ID NO:2 or 3.
8. The method of claim 1 , wherein the recombinant recognition molecule comprises a CDRH3 region that comprises the amino acid sequence of any one of SEQ ID NO: 4-6.
9. The method of claim 1 , wherein the CDRL1 region comprises the amino acid sequence of any one of SEQ ID NO: 7-9.
10. The method of claim 1 , wherein the CDRL1 region variant is selected from (a) a variant that has P substituted for S at position 2 of SEQ ID NO: 7, 8 or 9; (b) a variant that has F substituted for L at position 15 of SEQ ID NO: 7, 8 or 9; and (c) a variant that has P substituted for S at position 2 of SEQ ID NO: 7, 8 or 9 and F substituted for L at position 15 of SEQ ID NO: 7, 8 or 9.
11. The method of claim 1 , wherein the CDRL2 region comprises the amino acid sequence of any one of SEQ ID NO: 10-11.
12. The method of claim 1 , wherein the CDRL3 region comprises the amino acid sequence of any one of SEQ ID NO: 12-13.
13. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRH1 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
1 Q or E
2 V
3 Q, K or T
4 L
5 K or V
6 E or Q
7 S
8 G
9 A
10 E
11 L or V
12 V or K
13 R or K
14 P
15 G
16 T or A
17 S
18 V
19 K
20 I or V
21 S or P
22 C
23 K
24 A, V, S or T
25 S
26 G
27 Y, F, S or D
28 T
29 F, L or I
30 T.
14. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRH2 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
36 W
37 V
38 K or R
39 Q
40 R or A
41 P
42 G
43 H or Q
44 G
45 L
46 E
47 W or R
48 I or M
49 G.
15. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRH3 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
66 K or R
67 A or V
68 T
69 L or M
70 T
71 A, L or T
72 D
73 T
74 S
75 S or T
76 S
77 T
78 A
79 Y
80 M
81 Q or E
82 L
82a S
82b S or R
82c L
83 T or R
84 S
85 E
86 D
87 S or T
88 A
89 V
90 Y
91 F or Y
92 C
93 A
94 Y, K or R.
16. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRH4 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
103 W
104 G
105 Q
106 G
107 T
108 T, S or L
109 V or L
110 T
111 V
112 S
113 S or A.
17. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRL1 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
1 D
2 I, V or L
3 Q or L
4 M
5 T
6 Q
7 T or S
8 P
9 L
10 S
11 L
12 P
13 V
14 S or T
15 L or P
16 G
17 D or E
18 Q or P
19 A
20 S
21 I
22 S
23 C.
18. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRL2 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
35 W
36 Y
37 L
38 Q
39 K
40 P
41 G
42 Q
43 S
44 P
45 K or Q
46 L
47 L
48 I or V
49 Y.
19. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRL3 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
57 G
58 V
59 P
60 D
61 R
62 F
63 S
64 G
65 S
66 G
67 S
68 G
69 T
70 D
70 F
72 T
73 L
74 K
75 I
76 S
77 R
78 V
79 E
80 A
81 E
82 D
83 L or V
84 G
85 V
86 Y
87 Y
88 C.
20. The method of claim 1 , wherein the recombinant recognition molecule comprises an FRL4 region comprising the following sequence, the amino acid position corresponding to the numbering according to Kabat,
98 F
99 G
100 G or Q
101 G
102 T
103 K
104 L
105 E
106 I or L
106a K
107 R
108A.
21. The method of claim 1 wherein the recombinant recognition molecule comprises:
a. a CDRH1 region comprising the amino acid sequence SEQ ID NO: 1,
b. a CDRH2 region comprising the amino acid sequence SEQ ID NO: 2 or 3,
c. a CDRH3 region comprising the amino acid sequence SEQ ID NO: 4, 5, or 6,
d. a CDRL1 region comprising the amino acid sequence SEQ ID NO: 7, 8 or 9,
e. a CDRL2 region comprising the amino acid sequence SEQ ID NO: 10 or 11, and
f. a CDRL3 region comprising the amino acid sequence SEQ ID NO: 12 or 13.
22. A molecule for tumor recognition that is obtained by the method of any one of claims 1 - 21 .