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.

We claim: 
     
         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  or  2 , 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 any of  claims 1 - 3 , 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 any of  claims 1 - 3 , 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 any of  claims 1 - 3 , 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 any of the  claims 1 - 6 , wherein the Fc domain-containing binding proteins in the population comprises an antigen-binding portion of an antibody. 
     
     
         8 . The composition of any of the  claims 1 - 6 , wherein the Fc domain-containing binding proteins in the population comprise a non-antibody antigen-binding portion. 
     
     
         9 . The composition of  claim 7  or  8 , wherein the antigen-binding portion binds to tumor necrosis factor alpha (TNFα). 
     
     
         10 . The composition of  claim 9 , wherein the population of Fc domain-containing binding proteins comprises the polypeptide sequence of etanercept, infliximab, adalimumab, or golimumab. 
     
     
         11 . The composition of  claim 10 , wherein the population of Fc domain-containing binding proteins comprises the polypeptide sequence of adalimumab or a variant thereof. 
     
     
         12 . The composition of  claim 11 , wherein the variant of adalimumab exhibits pH-sensitive binding to the TNF antigen. 
     
     
         13 . The composition of  claim 12 , 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. 
     
     
         14 . The composition of any of  claims 11 - 13 , wherein the variant of adalimumab comprises a variant Fc region. 
     
     
         15 . The composition of  claim 14 , wherein the variant Fc region is a human IgG1 Fc region comprising the mutations T250Q and M428L relative to a wild-type human IgG1 sequence (numbering according to the EU convention as in Kabat). 
     
     
         16 . The composition of any of  claims 1 - 9 , wherein the Fc binding polypeptide is a dual variable domain immunoglobulin (DVD-Ig). 
     
     
         17 . The composition of any of  claims 1 - 16 , wherein the composition is obtained from a cultured mammalian host cell line. 
     
     
         18 . The composition of  claim 17 , wherein the host cell line is a CHO cell line containing a heterologous galactosyltransferase gene. 
     
     
         19 . The composition of  claim 17  or  18 , 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 any of the  claims 1 - 19  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. 
     
     
         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  or  22 , 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 any of the  claims 21 - 23 , 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 any of the  claims 21 - 24 , 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 any of the  claims 21 - 24 , 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 . The method of any of the  claims 21 - 26 , wherein the Fc domain-containing binding proteins bind to tumor necrosis factor alpha (TNFα). 
     
     
         28 . The method of  claim 27 , wherein the first population of Fc domain-containing binding proteins comprises etanercept, infliximab, adalimumab, or golimumab. 
     
     
         29 . The method of  claim 28 , wherein the first population of Fc domain-containing binding proteins comprises adalimumab or a variant thereof. 
     
     
         30 . The method of  claim 29 , wherein the variant of adalimumab exhibits pH-sensitive binding to the TNF antigen. 
     
     
         31 . The method of  claim 30 , 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. 
     
     
         32 . The method of any of  claims 29 - 31 , wherein the variant of adalimumab comprises a variant Fc region. 
     
     
         33 . The method of  claim 32 , wherein the variant Fc region is a human IgG1 Fc region comprising the mutations T250Q and M428L relative to a wild-type human IgG1 sequence (numbering according to the EU convention as in Kabat). 
     
     
         34 . The method of any of  claim 21 - 33 , 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. 
     
     
         35 . The method of  claim 34 , wherein the host cell line is a CHO cell line containing a heterologous galactosyltransferase gene. 
     
     
         36 . The method of  claim 34 , wherein the host cell line is a CHO cell line containing a knockdown of one or both the alleles of a Beta galactosidase gene. 
     
     
         37 . A host cell that produces a glycoengineered population of Fc domain-containing binding proteins wherein the population has one or more of the following properties:
 a) the total percent amount of G1 and G2 glycoforms in the population is more than 50%, more than 80%, or more than 99%;   b) less than 10%, less than 5%, less than 1%, or less than 0.1% of the Fc domain-containing binding proteins in the population comprise M3-M9 glycoforms;   c) a G1/2:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1;   d) a GS:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1; and   e) a Gtotal:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1.   
     
     
         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  claims 1 - 20 . 
     
     
         39 . The method of  claim 38 , wherein the disorder is a TNFα associated disorder.