Alteration of FcRn binding affinities or serum half-lives of antibodies by mutagenesis
View Patent ↗The present invention provides for a modified antibody of class IgG, in which at least one amino acid from the heavy chain constant region selected from the group consisting of amino acid residues 250, 314, and 428 is substituted with another amino acid which is different from that present in the unmodified antibody, thereby altering the binding affinity for FcRn and/or the serum half-life in comparison to the unmodified antibody.
1. A modified antibody of class IgG with FcRn binding affinity and/or serum half-life increased relative to that of an unmodified antibody, the modified antibody comprising a variable region from daclizumab and a heavy chain constant region, wherein the heavy chain constant region comprises glutamic acid or glutamine at amino acid residue 250 and leucine or phenylalanine at amino acid residue 428, EU numbering, and wherein
the variable region from daclizumab comprises a mature light chain variable region of SEQ ID NO:118 and a mature heavy chain variable region of SEQ ID NO: 122.
2. The modified antibody according to claim 1 , wherein said heavy chain constant region is that of IgG1.
3. An antibody comprising a variable region from daclizumab and a heavy chain constant region identical to that of a naturally occurring class IgG antibody except wherein
amino acid residue 250, EU numbering, is glutamic acid or glutamine and amino acid residue 428, EU numbering, is leucine or phenylalanine and different from residues 250 and 428 as present in the naturally occurring class IgG antibody,
the FcRn binding affinity and/or serum half-life of said antibody is altered relative to the naturally occurring antibody, and
the variable region from daclizumab comprises a mature light chain variable region of SEQ ID NO: 118 and a mature heavy chain variable region of SEQ ID NO: 122.
4. An antibody comprising a variable region from daclizumab and a heavy chain constant region, wherein:
(a) amino acid residue 250 from the heavy chain constant region is glutamic acid or glutamine;
(b) amino acid residue 428 from the heavy chain constant region is phenylalanine or leucine; and
(c) the variable region from daclizumab comprises a mature light chain variable region of SEQ ID NO:118 and a mature heavy chain variable region of SEQ ID NO: 122.
5. The antibody according to claim 3 , wherein said amino acid residue 250 from the heavy chain constant region is glutamine.
6. The antibody according to claim 3 , wherein said amino acid residue 428 from the heavy chain constant region is leucine.
7. The antibody according to claim 3 , wherein:
said amino acid residue 250 from the heavy chain constant region is glutamic acid and said amino acid residue 428 from the heavy chain constant region is phenylalanine;
said amino acid residue 250 from the heavy chain constant region is glutamine and said amino acid residue 428 from the heavy chain constant region is phenylalanine; or
said amino acid residue 250 from the heavy chain constant region is glutamine and said amino acid residue 428 from the heavy chain constant region is leucine.
8. A modified antibody of claim 1 with an in vivo clearance at least about 1.3-fold lower than that of the corresponding unmodified class IgG antibody.
9. A modified therapeutic antibody comprising a light chain amino acid sequence of SEQ ID NO: 118 and a heavy chain amino acid sequence selected from SEQ ID NOs: 122-123, and 127-128.
10. The modified therapeutic antibody of claim 9 comprising a light chain amino acid sequence of SEQ ID NO: 118 and a heavy chain amino acid sequence of SEQ ID NO: 122.
11. The modified therapeutic antibody of claim 9 comprising a light chain amino acid sequence of SEQ ID NO: 118 and a heavy chain amino acid sequence of SEQ ID NO: 127.
12. The modified therapeutic antibody of claim 9 comprising a light chain amino acid sequence of SEQ ID NO: 118 and a heavy chain amino acid sequence of SEQ ID NO: 123.
13. The modified therapeutic antibody of claim 9 comprising a light chain amino acid sequence of SEQ ID NO: 118 and a heavy chain amino acid sequence of SEQ ID NO: 128.
14. The modified antibody of claim 1 wherein said class IgG antibody is a human IgG1.
15. The modified antibody of claim 1 wherein said class IgG antibody is a human IgG2M3.
16. The antibody of claim 3 wherein said class IgG antibody is a human IgG1.