Fc-region variants with modified fcrn- and maintained protein a-binding properties

Abstract

Herein is reported a polypeptide comprising a first polypeptide and a second polypeptide each comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain, wherein i) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations H310A, H433A and Y436A, or ii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251D, L314D and L432D, or iii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251S, L314S and L432S.

1 . A polypeptide comprising
 a first polypeptide and a second polypeptide each comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain,   wherein
 i) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations H310A, H433A and Y436A, or 
 ii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251D, L314D and L432D, or 
 iii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251S, L314S and L432S. 
   
     
     
         2 . The polypeptide according to  claim 1 , wherein the polypeptide does not specifically bind to the human FcRn and does specifically bind to Staphylococcal protein A. 
     
     
         3 . The polypeptide according to  claim 1 , wherein i) the first polypeptide further comprises the mutations Y349C, T366 S, L368A and Y407V and the second polypeptide comprises the mutations S354C and T366W, or ii) the first polypeptide further comprises the mutations S354C, T366 S, L368A and Y407V and the second polypeptide comprises the mutations Y349C and T366W. 
     
     
         4 . The polypeptide according to  claim 1 , wherein the immunoglobulin hinge region, the immunoglobulin CH2-domain and the immunoglobulin CH3-domain are of the human IgG1 subclass. 
     
     
         5 . The polypeptide according to  claim 1 , wherein the first polypeptide and the second polypeptide further comprise the mutations L234A and L235A. 
     
     
         6 . The polypeptide according to  claim 1 , wherein the immunoglobulin hinge region, the immunoglobulin CH2-domain and the immunoglobulin CH3-domain are of the human IgG2 subclass. 
     
     
         7 . The polypeptide according to  claim 1 , wherein the immunoglobulin hinge region, the immunoglobulin CH2-domain and the immunoglobulin CH3-domain are of the human IgG4 subclass. 
     
     
         8 . The polypeptide according to  claim 1 , wherein the first polypeptide and the second polypeptide further comprise the mutations S228P and L235E. 
     
     
         9 . The polypeptide according to  claim 1 , wherein the first polypeptide and the second polypeptide further comprise the mutation P329G. 
     
     
         10 . The polypeptide according to  claim 1 , wherein the polypeptide is a bispecific antibody. 
     
     
         11 . A polypeptide comprising:
 a first polypeptide comprising in N-terminal to C-terminal direction a first heavy chain variable domain, an immunoglobulin CH1-domain of the subclass IgG1, an immunoglobulin hinge region of the subclass IgG1, an immunoglobulin CH2-domain of the subclass IgG1 and an immunoglobulin CH3-domain of the subclass IgG1,   a second polypeptide comprising in N-terminal to C-terminal direction a second heavy chain variable domain, an immunoglobulin CH1-domain of the subclass IgG1, an immunoglobulin hinge region of the subclass IgG1, an immunoglobulin CH2-domain of the subclass IgG1 and an immunoglobulin CH3-domain of the subclass IgG1,   a third polypeptide comprising in N-terminal to C-terminal direction a first light chain variable domain and a light chain constant domain,   a fourth polypeptide comprising in N-terminal to C-terminal direction a second light chain variable domain and a light chain constant domain,   wherein the first heavy chain variable domain and the first light chain variable domain form a first binding site that specifically binds to a first antigen,   wherein the second heavy chain variable domain and the second light chain variable domain form a second binding site that specifically binds to a second antigen,   wherein i) the first polypeptide comprises the mutations Y349C, T366 S, L368A and Y407V and the second polypeptide comprises the mutations S354C and T366W, or ii) the first polypeptide comprises the mutations S354C, T366 S, L368A and Y407V and the second polypeptide comprises the mutations Y349C and T366W,   wherein the first and the second polypeptide further comprise the mutations L234A, L235A and P329G, and   wherein
 i) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations H310A, H433A and Y436A, or 
 ii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251D, L314D and L432D, or 
 iii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251S, L314S and L432S. 
   
     
     
         12 . A polypeptide comprising:
 a first polypeptide comprising in N-terminal to C-terminal direction a first heavy chain variable domain, an immunoglobulin light chain constant domain, an immunoglobulin hinge region of the subclass IgG1, an immunoglobulin CH2-domain of the subclass IgG1 and an immunoglobulin CH3-domain of the subclass IgG1,   a second polypeptide comprising in N-terminal to C-terminal direction a second heavy chain variable domain, an immunoglobulin CH1-domain of the subclass IgG1, an immunoglobulin hinge region of the subclass IgG1, an immunoglobulin CH2-domain of the subclass IgG1 and an immunoglobulin CH3-domain of the subclass IgG1,   a third polypeptide comprising in N-terminal to C-terminal direction a first light chain variable domain and an immunoglobulin CH1-domain of the subclass IgG1,   a fourth polypeptide comprising in N-terminal to C-terminal direction a second light chain variable domain and a light chain constant domain,   wherein the first heavy chain variable domain and the first light chain variable domain form a first binding site that specifically binds to a first antigen,   wherein the second heavy chain variable domain and the second light chain variable domain form a second binding site that specifically binds to a second antigen,   wherein i) the first polypeptide comprises the mutations Y349C, T366 S, L368A and Y407V and the second polypeptide comprises the mutations S354C and T366W, or ii) the first polypeptide comprises the mutations S354C, T366 S, L368A and Y407V and the second polypeptide comprises the mutations Y349C and T366W,   wherein the first and the second polypeptide further comprise the mutations L234A, L235A and P329G, and   wherein
 i) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations H310A, H433A and Y436A, or 
 ii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251D, L314D and L432D, or 
 iii) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251S, L314S and L432S. 
   
     
     
         13 . A polypeptide according to  claims 1 ,  11  or  12  for intravitreal application. 
     
     
         14 . A polypeptide according to  claims 1 ,  11  or  12  for the treatment of vascular eye diseases. 
     
     
         15 . A pharmaceutical formulation comprising a polypeptide according to  claims 1 ,  11  or  12  and optionally a pharmaceutically acceptable carrier. 
     
     
         16 .- 18 . (canceled) 
     
     
         19 . A method for the transport of a soluble receptor ligand from the eye over the blood-ocular-barrier into the blood circulation in an individual comprising administering to the individual an effective amount of the polypeptide according to  claim 1 ,  11  or  12 . 
     
     
         20 . A method for the removal of one or more soluble receptor ligands from the eye in an individual comprising administering to the individual an effective amount of the polypeptide according to  claim 1 ,  11  or  12 . 
     
     
         21 . A method for treatment of an eye disease with the polypeptide according to  claim 1 ,  11  or  12 .