Aspgr antibodies and uses thereof

Abstract

The present invention generally relates to antibodies specific for asialoglycoprotein receptor (ASGPR) and their use for selectively delivering effector moieties that influence cellular activity. In addition, the present invention relates to polynucleotides encoding such antibodies, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the antibodies of the invention, and to methods of using them in the treatment of disease.

1 - 24 . (canceled) 
     
     
         25 . A method for producing an antibody capable of specific binding to asialoglycoprotein (ASGPR), comprising the steps of:
 (a) culturing a host cell comprising:
 (i) a polynucleotide encoding the antibody or an antigen binding portion thereof, wherein the antibody competes with any of antibodies comprising:
 a) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 2; 
 b) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 30; 
 c) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 32; 
 d) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 34; 
 e) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 36; 
 f) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 38; 
 g) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 40; 
 h) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 42; 
 i) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 44; 
 i) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 46; or 
 j) a heavy chain variable region sequence of SEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 48; 
 
 for binding to an epitope of ASGPR, or 
 (ii) an expression vector, comprising the polynucleotide of step (i) under conditions for expression of said antibody, and 
   (b) recovering said antibody.   
     
     
         26 . The method of  claim 25 , wherein the antibody comprises the heavy chain variable region sequence of SEQ ID NO: 4 and the light chain variable region sequence of SEQ ID NO: 2. 
     
     
         27 . The method of  claim 25 , wherein the antibody is an affinity matured variant of any of the antibodies comprising a)-j). 
     
     
         28 . The method of  claim 25 , wherein the antibody is capable of specific binding to human and cynomolgus monkey ASGPR. 
     
     
         29 . The method of  claim 28 , wherein the antibody comprises a human Fc region. 
     
     
         30 . The method of  claim 29 , wherein the antibody comprises a modification within the interface between the two antibody heavy chains in the CH3 domain, wherein i) in the CH3 domain of one heavy chain, an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance (“knob”) within the interface in the CH3 domain of one heavy chain which is positionable in a cavity (“hole”) within the interface in the CH3 domain of the other heavy chain, and ii) in the CH3 domain of the other heavy chain, an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity (“hole”) within the interface in the second CH3 domain within which a protuberance (“knob”) within the interface in the first CH3 domain is positionable. 
     
     
         31 . The method of  claim 29 , wherein the antibody comprises the amino acid substitution T366W and optionally the amino acid substitution S354C in one of the antibody heavy chains, and the amino acid substitutions T366S, L368A, Y407V and optionally Y349C in the other of the antibody heavy chains. 
     
     
         32 . The method of  claim 28 , wherein the antibody is a full-length antibody. 
     
     
         33 . The method of  claim 28 , wherein the antibody comprises an Fc region modification that reduces binding affinity of the antibody to an Fc receptor, or an Fcγ receptor; wherein said Fc receptor is an activating Fc receptor and wherein the activating Fc receptor is FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), or FcαRI (CD89). 
     
     
         34 . The method of  claim 33 , wherein the Fc receptor is FcγRIIIa, or human FcγRIIIa. 
     
     
         35 . The method of  claim 33 , wherein the antibody comprises an amino acid substitution in the Fc region at a position selected from the group consisting of P329, L234 and L235 (EU numbering). 
     
     
         36 . The method of  claim 33 , wherein the antibody comprises the amino acid substitutions P329G, L234A and L235A in the Fc region (EU numbering). 
     
     
         37 . The method of  claim 28 , wherein the antibody comprises an effector moiety attached to the antibody, wherein the effector moiety is a cytokine molecule. 
     
     
         38 . The method of  claim 37 , wherein no more than one effector moiety is attached to the antibody. 
     
     
         39 . The method of  claim 37 , wherein the cytokine molecule is fused by an amino-terminal amino acid to a carboxy-terminal amino acid of one of the heavy chains of the antibody. 
     
     
         40 . The method of  claim 37 , wherein the cytokine molecule is a human interferon alpha molecule. 
     
     
         41 . The method of  claim 37 , wherein the cytokine molecule is an interferon molecule selected from the group consisting of human interferon alpha 2 and human interferon alpha 2a. 
     
     
         42 . The method of  claim 28 , wherein the antibody comprises an IgG Fc region or an IgG1 Fc region. 
     
     
         43 . The method of  claim 28 , wherein the antibody is an IgG class antibody or an IgG subclass antibody.