Method of screening antibodies with high antigen selectivity

1. A method of screening antibodies, the method comprising:

contacting an antigen protein with a solution of calcium or calcium salt, wherein the antigen protein is a truncated version of a wild-type protein of interest;

performing a negative selection on an antibody library, wherein the negative selection comprises contacting the library with a full-length version of the wild type protein of interest and removing any antibody bound to the full-length version of the wild type protein of interest from the antibody library;

contacting the antibody library with the antigen protein in the presence of the calcium or calcium salt; and

selecting one or more antibody or antibody fragments from the antibody library with the highest binding affinity to the antigen protein.

2. The method of claim 1 , wherein the antigen protein is a truncated version of a wild-type protein selected from the group consisting of Annexin, calmodulin, EF hand protein, and C2 domain protein.

3. The method of claim 2 , wherein the antigen protein is a truncated version of Annexin A1 protein.

4. The method of claim 3 , wherein the Annexin A1 protein has an amino acid sequence set forth in SEQ ID NO: 1.

5. The method of claim 1 , wherein the antigen protein is an Annexin A1 protein with a truncated N-terminal or C-terminal.

6. The method of claim 2 , wherein the Annexin has an N-terminal that is truncated.

7. The method of claim 6 , wherein the Annexin having the N-terminal that is truncated has an amino acid sequence set forth in SEQ ID NO: 2.

8. The method of claim 1 , wherein the antibody library is a phage display library.

9. The method of claim 5 , wherein the negative selection comprises contacting the antibody library with an Annexin full-length protein, and removing an antibody bound to the Annexin full-length protein from the antibody library.