Dual specific antibodies for binding interleukin 4 (IL4) and interleukin 5 (IL5)
The invention provides dual specific antibodies capable of binding interleukin 4 (IL4) and interleukin 5 (IL5) and methods of making and using such antibodies. In general, the dual specific antibodies capable of binding IL4 and IL5 are generated by identification of a monospecific antibody having light chain variable region V L residues that are electrostatic or hydrophobic and altering the nucleic acid sequence encoding one or more solvent accessible residues in the V H of the antibody either alone or in combination with alteration of the nucleic acid sequence encoding the V L of the antibody. The altered V H and the V L are expressed and dual specific antibodies, or antigen-binding fragments thereof, are selected.
1. An isolated dual specific antibody, or antigen-binding fragment thereof, capable of binding interleukin 4 (IL4) and interleukin 5 (IL5), comprising the following six CDRs:
(i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9);
(ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10);
(iii) a CDRL3 comprising the amino acid sequence of QQDYKHPWTF (SEQ ID NO: 31);
(iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15);
(v) a CDRH2 comprising the amino acid sequence of AGIVYDATGFTVYADDFK (SEQ ID NO: 32); and
(vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).
2. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , further comprising a framework region 3 (FR3) comprising the amino acid sequence of GRX 1 TITX 2 DX 3 STSTX 4 (SEQ ID NO: 26), wherein X 1 is Val or Phe, X 2 is Arg or Ile, X 3 is Thr, Phe, Met, or Pro, and X 4 is Ala or Val.
3. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody binds IL4 with a Kd of 10 nM or lower and IL5 with a Kd of about 50 nM or lower.
4. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody is an IgG antibody.
5. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , wherein said fragment is a Fab fragment or a single chain variable fragment (scFv).
6. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , wherein at least a portion of the framework sequence is a human consensus framework sequence.
7. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , wherein the antibody is a chimeric antibody.
8. The isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , wherein the antibody is a humanized antibody.
9. A pharmaceutical composition comprising the isolated dual specific antibody, or antigen-binding fragment thereof, of claim 1 , and a pharmaceutically acceptable excipient.
10. An isolated dual specific antibody, or antigen-binding fragment thereof, capable of binding IL4 and IL5, comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 33 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 34.
11. A pharmaceutical composition comprising the isolated dual specific antibody, or antigen-binding fragment thereof, of claim 10 , and a pharmaceutically acceptable excipient.