Humanized anti-MUC1* antibodies
The present application discloses humanized antibodies and antibody like proteins and fragments thereof.
1. A chimeric antigen receptor (CAR) comprising
(a) an anti-MUC1* antibody fragment wherein the anti-MUC1* antibody fragment comprises a heavy chain variable region and a light chain variable region that has complementarity determining regions (CDRs) in the heavy chain variable region and the light chain variable region as follows:
CDR1 heavy chain SEQ ID NO: 123,
CDR1 light chain SEQ ID NO: 173,
CDR2 heavy chain SEQ ID NO: 127,
CDR2 light chain SEQ ID NO: 177,
CDR3 heavy chain SEQ ID NO: 131, and
CDR3 light chain SEQ ID NO: 181; and
(b) a T cell receptor co-stimulatory domain.
2. The chimeric antigen receptor of claim 1 , wherein the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 145 and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 195.
3. The chimeric antigen receptor of claim 1 , wherein the heavy chain variable region comprises an amino acid sequence that has at least 95% sequence identity to SEQ ID NO: 145 and the light chain variable region comprises an amino acid sequence that has at least 95% sequence identity to SEQ ID NO: 195.
4. The chimeric antigen receptor of claim 1 , wherein the heavy chain variable region comprises an amino acid sequence that has at least 98% sequence identity to SEQ ID NO: 145 and the light chain variable region comprises an amino acid sequence that has at least 98% sequence identity to SEQ ID NO: 195.
5. The chimeric antigen receptor of claim 1 , wherein the heavy chain variable region comprises the amino acid sequence according to SEQ ID NO: 145 and the light chain variable region comprises the amino acid sequence according to SEQ ID NO: 195.
6. The chimeric antigen receptor of claim 1 , wherein the heavy chain variable region and the light chain variable region are connected by a linker.
7. The chimeric antigen receptor of claim 6 , wherein the linker comprises the amino acid sequence according to SEQ ID NO: 402.
8. The chimeric antigen receptor of claim 1 , wherein the anti-MUC1* antibody fragment comprises a single chain variable fragment (scFv).
9. The chimeric antigen receptor of claim 8 , wherein the scFv comprises an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 239, 241, 243, or 398.
10. The chimeric antigen receptor of claim 8 , wherein the scFv comprises an amino acid sequence that has at least 95% sequence identity to any one of SEQ ID NOs: 239, 241, 243, or 398.
11. The chimeric antigen receptor of claim 8 , wherein the scFv comprises an amino acid sequence that has at least 98% sequence identity to any one of SEQ ID NOs: 239, 241, 243, or 398.
12. The chimeric antigen receptor of claim 8 , wherein the scFv comprises an amino acid sequence according to any one of SEQ ID NOs: 239, 241, 243, or 398.
13. The chimeric antigen receptor of claim 1 , wherein the T cell receptor co-stimulatory domain comprises a cytoplasmic domain of CD3-zeta, CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICAM-1, LFA-1, ICOS, CD2, CD5, CD7, or Fc receptor gamma domain.
14. The chimeric antigen receptor of claim 1 , wherein the T cell receptor co-stimulatory domain comprises a cytoplasmic domain of CD3-zeta.
15. The chimeric antigen receptor of claim 1 , wherein the T cell receptor co-stimulatory domain comprises a cytoplasmic domain of 4-1BB.
16. The chimeric antigen receptor of claim 1 , wherein the T cell receptor co-stimulatory domain comprises a cytoplasmic domain of CD3-zeta and a cytoplasmic domain of 4-1BB.
17. The chimeric antigen receptor of claim 16 , wherein the cytoplasmic domain of CD3-zeta comprises an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 374.
18. The chimeric antigen receptor of claim 16 , wherein the cytoplasmic domain of 4-1BB comprises an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 380.
19. The chimeric antigen receptor of claim 16 , wherein the T cell receptor co-stimulatory domain comprises the amino acid sequences of SEQ ID NO: 374 and SEQ ID NO: 380.
20. The chimeric antigen receptor of claim 16 , wherein the CAR comprises an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 611.
21. The chimeric antigen receptor of claim 16 , wherein the CAR comprises an amino acid sequence that has at least 95% sequence identity to SEQ ID NO: 611.
22. The chimeric antigen receptor of claim 16 , wherein the CAR comprises an amino acid sequence that has at least 98% sequence identity to SEQ ID NO: 611.
23. The chimeric antigen receptor of claim 8 , wherein the scFv binds to a MUC1* peptide, wherein the MUC1* peptide comprises the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 620, or SEQ ID NO: 621.
24. The chimeric antigen receptor of claim 8 , wherein the scFv binds to a MUC1* positive cancer cell or a MUC1* transfected cell.
25. The chimeric antigen receptor of claim 24 , wherein the MUC1* positive cancer cell is a MUC1* positive breast cancer cell.
26. The chimeric antigen receptor of claim 24 , wherein the MUC1* positive cancer cell is a MUC1* positive prostate cancer cell.
27. The chimeric antigen receptor of claim 8 , wherein the scFv inhibits growth of a MUC1* positive cancer cell.
28. The chimeric antigen receptor of claim 27 , wherein the MUC1* positive cancer cell is a MUC1* positive breast cancer cell.
29. A cell comprising the CAR of claim 1 .
30. The cell of claim 29 , wherein the cell comprises a T cell, a dendritic cell, or a mast cell.