Alteration of Fc-fusion protein serum half-lives by mutagenesis
View Patent ↗The present invention provides for a modified Fc-fusion protein 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 Fc-fusion protein, thereby altering the binding affinity for FcRn and/or the serum half-life in comparison to the unmodified Fc-fusion protein.
1. A polynucleotide encoding an FcRN binding portion of an IgG constant region comprising amino acid residues 250 and 428 that differ from the residues present in an unmodified FcRN binding portion of an IgG constant region by amino acid residue 250 being glutamic acid or glutamine and amino acid residue 428 being leucine or phenylalanine, wherein the amino acid residues are numbered by the EU numbering system.
2. The polynucleotide of claim 1 , wherein the IgG isotype is a human IgG isotype.
3. The polynucleotide of claim 1 wherein the IgG isotype is human IgG1.
4. The polynucleotide of claim 1 wherein amino acid residue 250 is glutamine and amino acid residue 428 is leucine.
5. The polynucleotide of claim 1 , wherein the polynucleotide encoding an FcRN binding portion is linked in-frame to a bioactive protein.
6. The polynucleotide of claim 5 , wherein the FcRN binding portion is linked in-frame to the bioactive protein by a linker and hinge region.
7. The polynucleotide of claim 1 that is a component of an expression vector and operably linked to control sequences in the expression vector.