Light chain variable regions
Novel modulators, including antibodies and derivatives thereof, and methods of using such modulators to treat proliferative disorders are provided.
1. A polypeptide comprising a set of three non-contiguous sequences of amino acids, wherein the amino acids of the set of three non-contiguous sequences are:
(1) TASSSVSSSY (SEQ ID NO: 427);
(2) STSNLAS (SEQ ID NO: 428), wherein the serine residue at position 1 of SEQ ID NO: 428 is optionally substituted by another amino acid and the asparagine residue at position 4 of SEQ ID NO: 428 is optionally substituted by another amino acid; and
(3) HQX 1 X 2 RSPX 3 TFGX 4 GTKLX 5 X 6 X 7 (SEQ ID NO: 429), wherein X 1 is F or Y; X 2 is H or N; X 3 is L or F; X 4 is A or S; X 5 is E or K; X 6 is L or I; and X 7 is K or R.
2. The polypeptide of claim 1 , wherein the amino acid at position 1 of SEQ ID NO: 428 is a charged amino acid.
3. The polypeptide of claim 1 , wherein the amino acid at position 1 of SEQ ID NO: 428 is selected from the group consisting of lysine, arginine, histidine, aspartic acid, and glutamic acid.
4. The polypeptide of claim 1 , wherein the amino acid at position 1 of SEQ ID NO: 428 is arginine.
5. The humanized light chain of claim 1 , wherein the amino acid at position 4 of SEQ ID NO: 428 is a charged amino acid.
6. The polypeptide of claim 1 , wherein the amino acid at position 4 of SEQ ID NO: 428 is selected from the group consisting of lysine, arginine, histidine, aspartic acid, and glutamic acid.
7. The polypeptide of claim 1 , wherein the amino acid at position 4 of SEQ ID NO: 428 is arginine.
8. A humanized light chain comprising a light chain variable region having three complementarity determining regions (CDRs) derived from a donor sequence, wherein the three CDRs comprise:
(1) TASSSVSSSY (SEQ ID NO: 427);
(2) STSNLAS (SEQ ID NO: 428), wherein the serine residue at position 1 of SEQ ID NO: 428 is optionally substituted by another amino acid and the asparagine residue at position 4 of SEQ ID NO: 428 is optionally substituted by another amino acid; and
(3) HQX 1 X 2 RSPX 3 T (SEQ ID NO: 430), wherein X 1 is F or Y; X 2 is H or N; and X 3 is L or F.
9. The humanized light chain of claim 8 , wherein the light chain variable region comprises four VK framework regions, wherein each of the VK framework regions are derived from a human framework sequence.
10. The humanized light chain of claim 9 , wherein the VK framework regions comprise a second non-contiguous amino acid sequence comprising:
EIVLTQSPGTLSLSPGERATLSC (FR1) (SEQ ID NO: 414), wherein the glutamic acid residue at position 1, the leucine residue at position 21, and the serine residue at position 22 of SEQ ID NO: 414 are each independently and optionally substituted by a residue from the donor sequence;
WYQQKPGQAPRLLIY (FR2) (SEQ ID NO: 416), wherein the leucine residue at position 13 of SEQ ID NO: 416 is optionally substituted by a residue from the donor sequence;
GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC (FR3) (SEQ ID NO: 418), wherein the isoleucine residue at position 2, the phenylalanine residue at position 27, and the valine residue at position 29 of SEQ ID NO: 418 are each independently and optionally substituted by a residue from the donor sequence; and
FGQGTKVEIK (FR4) (SEQ ID NO: 420), wherein the glutamine residue at position 3 and the valine residue at position 7 of SEQ ID NO: 420 are each independently and optionally substituted by a residue from the donor sequence.
11. The humanized light chain of claim 10 , wherein the amino acid at position 1 of SEQ ID NO: 414 is glutamine, the amino acid at position 21 of SEQ ID NO: 414 is methionine, isoleucine, or leucine, and the amino acid at position 22 of SEQ ID NO: 414 is threonine.
12. The humanized light chain of claim 10 , wherein the amino acid at position 13 of SEQ ID NO: 416 is tryptophan.
13. The humanized light chain of claim 10 , wherein the amino acid at position 2 of SEQ ID NO: 418 is valine, the amino acid at position 27 of SEQ ID NO: 418 is alanine, and the amino acid at position 29 of SEQ ID NO: 418 is threonine.
14. The humanized light chain of claim 10 , wherein the amino acid at position 3 of SEQ ID NO: 420 is alanine or serine and the amino acid at position 7 of SEQ ID NO: 420 is leucine.
15. The humanized light chain of claim 10 , wherein the amino acid at position 1 of SEQ ID NO: 428 is a charged amino acid and the amino acid at position 4 of SEQ ID NO: 428 is a charged amino acid.
16. The humanized light chain of claim 15 , wherein the amino acid at position 1 of SEQ ID NO: 414 is glutamine, the amino acid at position 21 of SEQ ID NO: 414 is methionine, isoleucine, or leucine, and the amino acid at position 22 of SEQ ID NO: 414 is threonine; the amino acid at position 13 of SEQ ID NO: 416 is tryptophan; the amino acid at position 2 of SEQ ID NO: 418 is valine, the amino acid at position 27 of SEQ ID NO: 418 is alanine, and the amino acid at position 29 of SEQ ID NO: 418 is threonine; and the amino acid at position 3 of SEQ ID NO: 420 is alanine or serine and the amino acid at position 7 of SEQ ID NO: 420 is leucine.