Glycoengineered binding protein compositions

Abstract

Provided are glycoengineered populations of Fc domain-containing binding proteins with a reduced anti-drug immune response (ADA). Also provided are methods of treating disease using such compositions, and methods and host for making such compositions.

1 . A glycoengineered binding protein composition comprising a population of Fc domain-containing binding proteins having an G/M ratio of at least 10:1, wherein the total percent amount of G1 and G2 glycoforms in the population is more than 50%, wherein the total percent amount of M3-M9 glycoforms is less than 10%, wherein Fc domain containing binding proteins of the composition comprise the same polypeptide sequence, and wherein the glycoengineered binding protein composition exhibits a lower ADA response and/or a greater serum half-life than a non-hypergalactosylated population of Fc domain-containing binding proteins. 
     
     
         2 . The composition of  claim 1 , wherein the total percent amount of G1 and G2 glycoforms in the population is more than 80%, or more than 99%. 
     
     
         3 . The composition of  claim 1 , wherein less than 5%, or less than 1%, or less than 0.1% of the Fc domain-containing binding proteins in the population comprise M3-M9 glycoforms. 
     
     
         4 . The composition of  claim 1 , wherein the population of Fc domain-containing binding proteins has a G1/2:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1. 
     
     
         5 . The composition of  claim 1 , wherein the population of Fc domain-containing binding proteins has a GS:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1. 
     
     
         6 . The composition of  claim 1 , wherein the population of Fc domain-containing binding proteins has a Gtotal:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1. 
     
     
         7 . The composition of  claim 1 , wherein the Fc domain-containing binding proteins in the population comprises an antigen-binding portion of an antibody or wherein the Fc domain-containing binding proteins in the population comprise a non-antibody antigen-binding portion, wherein the antigen-binding portion binds to tumor necrosis factor alpha (TNFα). 
     
     
         8 .- 10 . (canceled) 
     
     
         11 . The composition of  claim 7 , wherein the population of Fc domain-containing binding proteins comprises the polypeptide sequence of adalimumab or a variant thereof, wherein the variant of adalimumab is D2E7SS22 and comprises the heavy chain variable region sequence of SEQ ID NO:1 and the light chain variable region sequence of the light chain of SEQ ID NO:2. 
     
     
         12 .- 16 . (canceled) 
     
     
         17 . The composition of  claim 1 , wherein the composition is obtained from a cultured mammalian host cell line, wherein the host cell line is a CHO cell line containing a heterologous galactosyltransferase gene. 
     
     
         18 . (canceled) 
     
     
         19 . The composition of  claim 17 , wherein the host cell line is a CHO cell line containing a knockdown of one or both the alleles of a Beta galactosidase gene. 
     
     
         20 . A composition comprising the population of Fc domain-containing binding proteins of  claim 1  and a pharmaceutically acceptable carrier or excipient. 
     
     
         21 . A method of reducing a subject's anti-drug antibody (ADA) response to a first population of Fc domain-containing binding proteins, the method comprising glycoengineering the first population of Fc domain-containing binding proteins to obtain a binding protein composition comprising a second population of Fc-domain containing binding proteins having a G/M ratio of at least 10:1, a total percent amount of G1 and G2 glycoforms of more than 50%, and a total percent amount of M3-M9 glycoforms of less than 10%, wherein the second population of Fc domain-containing binding proteins has a greater serum half-life than the first population of Fc domain-containing binding proteins, wherein the Fc domain-containing binding proteins bind to tumor necrosis factor alpha (TNFα). 
     
     
         22 . The method of  claim 21 , wherein the total percent amount of G1 and G2 glycoforms in the second population is more than 80%, or more than 99%. 
     
     
         23 . The method of  claim 21 , wherein less than 5%, or less than 1%, or less than 0.1% of the Fc domain-containing binding proteins in the second population comprise M3-M9 glycoforms. 
     
     
         24 . The method of  claim 21 , wherein the second population of Fc domain-containing binding proteins has a G1/2:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1. 
     
     
         25 . The method of  claim 21 , wherein the second population of Fc domain-containing binding proteins has a GS:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1. 
     
     
         26 . The method of  claim 21 , wherein the second population of Fc domain-containing binding proteins has a Gtotal:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1. 
     
     
         27 .- 28 . (canceled) 
     
     
         29 . The method of  claim 21 , wherein the first population of Fc domain-containing binding proteins comprises adalimumab or a variant thereof, wherein the variant of adalimumab is D2E7SS22 and comprises the heavy chain variable region sequence of SEQ ID NO:1 and the light chain variable region sequence of the light chain of SEQ ID NO:2. 
     
     
         30 .- 33 . (canceled) 
     
     
         34 . The method of  claim 21 , wherein said glycoengineering comprises expressing the Fc-domain containing binding proteins in cultured mammalian host cell line that has been glycoengineered to produce hypergalactosylated and/or hypomannosylated binding proteins, wherein the host cell line is a CHO cell line containing a heterologous galactosyltransferase gene, wherein the host cell line is a CHO cell line containing a knockdown of one or both the alleles of a Beta galactosidase gene. 
     
     
         35 .- 37 . (canceled) 
     
     
         38 . A method of treating a disorder in a subject in need thereof, comprising administering to the subject an effective amount of the glycoengineered composition of any one of  claim 1 , wherein the disorder is a TNFα associated disorder. 
     
     
         39 . (canceled)