Anti-hepcidin antibodies and methods of use
The invention relates to monoclonal antibodies that bind hepcidin and methods of making and using such antibodies. Also provided are methods of treating hepcidin-related disorders.
1. An isolated antibody comprising SEQ ID NOs: 334-339 wherein the antibody binds to human hepcidin of SEQ ID NO: 9 with an affinity K D of less than about 10 −8 M and which exhibits at least one of the properties selected from the group consisting of:
(a) at least about a 50-fold higher K D at a pH of about 5.5 compared to its K D for said hepcidin at a pH of about 7.4;
(b) at least about a 5-fold faster clearance of said hepcidin compared to antibody 1S1 having the heavy chain variable region of SEQ ID NO: 202 and the light chain variable region of SEQ ID NO: 128; and
(c) an off rate of about 6×10 −2 s −1 or higher at about pH 5.5.
2. An isolated antibody comprising the amino acid sequence SEQ ID NOs: 334-339 that binds to human hepcidin of SEQ ID NO: 9, with an affinity K D of at least 10 −8 M and said antibody has at least about a 50-fold higher K D at about pH 5.5 compared to its K D for said hepcidin at about pH 7.4 or said antibody has an off rate of about 6×10 −2 s −1 or higher at about pH 5.5, wherein said antibody is obtained by:
(a) replacing an amino acid in the heavy or light chain of said antibody with a histidine;
(b) screening the antibody obtained in (a) for differential pH binding;
(c) replacing another amino acid in the heavy or light chain of said antibody with a histidine; and
(d) screening said antibody for having at least one of the properties selected from the group consisting of:
(i) at least about 50-fold higher K D at about pH 5.5 compared to its K D for said hepcidin at about pH 7.4; and
(ii) an off rate of about 6×10 −2 s −1 or higher at about pH 5.5.