Fc-receptor binding modified asymmetric antibodies and methods of use
Herein is reported an IgG class Fc-region comprising a first variant Fc-region polypeptide and a second variant Fc-region polypeptide, wherein the first variant Fc-region polypeptide is derived from a first parent IgG class Fc-region polypeptide and the second variant Fc-region polypeptide is derived from a second parent IgG class Fc-region polypeptide, whereby the first parent IgG class Fc-region polypeptide is identical to or different from the second parent IgG class Fc-region polypeptide, and the first variant Fc-region polypeptide differs from the second variant Fc-region polypeptide in one or more amino acid residues other than those amino acid residues in which the first parent IgG class Fc-region polypeptide differs from the second parent IgG class Fc-region polypeptide, and the IgG class Fc-region comprising the first variant Fc-region polypeptide and the second variant Fc-region polypeptide has an affinity to a human Fc-receptor that is different than that of an IgG class Fc-region comprising the first parent IgG class Fc-region polypeptide of a) and the second parent IgG class Fc-region polypeptide of a).
1 . An IgG class Fc-region comprising a first variant Fc-region polypeptide and a second variant Fc-region polypeptide, wherein:
a) the first variant Fc-region polypeptide is derived from a first parent IgG class Fc-region polypeptide and the second variant Fc-region polypeptide is derived from a second parent IgG class Fc-region polypeptide, whereby the first parent IgG class Fc-region polypeptide is identical to or different from the second parent IgG class Fc-region polypeptide;
b) the first variant Fc-region polypeptide differs from the second variant Fc-region polypeptide in one or more amino acid residues other than those amino acid residues in which the first parent IgG class Fc-region polypeptide differs from the second parent IgG class Fc-region polypeptide; and
c) the IgG class Fc-region comprising the first variant Fc-region polypeptide and the second variant Fc-region polypeptide has an affinity to a human Fc-receptor that is different than that of an IgG class Fc-region comprising the first parent IgG class Fc-region polypeptide of a) and the second parent IgG class Fc-region polypeptide of a),
wherein:
the human Fc-receptor is the human neonatal Fc-receptor;
the first and the second Fc-region polypeptide are both of human IgG1 of SEQ ID NO: 60 and have the following mutations (numbering according to Kabat EU index numbering system): H433A in the first variant Fc-region polypeptide, and H310A and Y436A in the second variant Fc-region polypeptide, or H433A and H310A in the first variant Fc-region polypeptide, and Y436A in the second variant Fc-region polypeptide, or H433A and Y436A in the first variant Fc-region polypeptide, and H310A in the second variant Fc-region polypeptide;
the IgG1 Fc-region has a reduced binding to Staphylococcus protein A than an IgG1 Fc-region comprising the first IgG1 Fc-region polypeptide of a) and the second IgG1 Fc-region polypeptide of a); and
wherein:
i) the first IgG1 Fc-region polypeptide and the second IgG1 Fc-region polypeptide have no mutations other than those stated above, or
ii) the first IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A and the second IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, or
iii) the first IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, 1235A, P329G and the second IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, or
iv) the first IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, S354C, T366W and the second IgG1 FC-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, Y349C, T366S, L368A, Y407V, or
v) the first IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, S354C, T366W and the second IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, Y349C, T366S, L368A, Y407V, or
vi) the first IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutation K392D and the second IgG1 Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations D399K, D356K, and/or E357K.
2 . The IgG class Fc-region according to claim 1 , wherein the human Fc-receptor is the human neonatal Fc receptor (FcRn) or the human Fcgamma III receptor (FcγRIII).
3 . The IgG class Fc-region according to claim 2 , wherein the human Fc-receptor is the human neonatal Fc-receptor.
4 . The IgG class Fc-region according to claim 1 , wherein the affinity to a human Fc-receptor is increased or reduced by 10% or more determined by surface plasmon resonance (SPR).