Humanized antibodies

1. An isolated humanized monoclonal antibody or antigen-binding fragment thereof that binds to a target antigen prepared by a method comprising,

(1) obtaining the sequence of a non-human donor antibody that binds to said target antigen, and determining the donor CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 sequences of said non-human donor antibody;

(2) obtaining the sequences of a human germline VL and a human germline VH, and determining the germline framework and germline CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 sequences of said human VL and VH;

(3) aligning each of the non-human donor CDR-L1, CDR-L2, and CDR-L3 sequences with the corresponding germline CDR sequence from said human VL, and each of the non-human CDR-H1 and CDR-H2 sequences with corresponding germline CDR sequence from said human VH;

(4) identifying positions in CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 where human germline residue is the same as, or different from, the corresponding non-human donor residue; and

(5) generating the isolated humanized monoclonal antibody or antigen-binding fragment thereof by changing a non-human donor residue of the donor CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 to the corresponding human germline residue if the human germline residue at the position is different from the corresponding non-human residue, wherein:

(a) said isolated humanized monoclonal antibody or antigen-binding fragment thereof comprises (i) a VH domain comprising a human germline VH framework sequence from step (2) and a VL domain comprising a human germline VL framework sequence from step (2); and (ii) a CDR-L1, CDR-L2, CDR-L3, CDR-H1 and CDR-H2 from step (5); and (iii) a CDR-H3;

(b) for each position within said CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2, the residue is either human germline residue from said human germline VL or VH, or corresponding residue from said non-human donor antibody;

(c) said CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 each comprises at least one more human germline residue as compared to the corresponding non-human donor CDR,

(d) said CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 each comprises at least one more non-human donor residue as compared to the corresponding human germline VH or VL CDR; and

(e) for each position within CDR-H3, the residue is any one of the 20 natural amino acid residues.

2. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence comprises a VH3, VH1, or VH5 framework sequence.

3. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence comprises a VH germline consensus framework sequence.

4. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence comprises the VH framework sequence of any one of the consensus sequences listed in Table 2 and Table 5.

5. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence comprises a VK or V?, framework sequence.

6. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence comprises a VL germline consensus framework sequence.

7. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence comprises the VL framework sequence of any one of the human germline sequences listed in Table 3, Table 4 and Table 6.

8. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence is derived from the VH framework sequence of human germline DP54.

9. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence is derived from the VH framework sequence of SEQ ID NO:74.

10. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence is derived from a VK framework sequence.

11. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence is derived from the VL framework sequence of human germline DPK9.

12. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence is derived from the VL framework sequence of SEQ ID NO:144.

13. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment binds said target antigen with a binding affinity (Kd) value that is equal or less than the binding affinity (Kd) value of said non-human donor antibody.

14. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment maintains highly specific binding to said target antigen as compared to binding of said non-human donor antibody to said target antigen.