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 is selected from the group consisting of:
 (1) a population of Fc domain-containing binding proteins having a G1/2:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1,   (2) a population of Fc domain-containing binding proteins having a GS:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1 and   (3) a population of Fc domain-containing binding proteins having a Gtotal:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1.   
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         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 a non-antibody antigen binding portion. 
     
     
         8 . (canceled) 
     
     
         9 . The composition of  claim 7 , 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 or the polypeptide sequence of a variant of etanercept, infliximab, adalimumab, or golimumab. 
     
     
         11 . (canceled) 
     
     
         12 . The composition of  claim 11 , wherein the variant of adalimumab is selected from the group consisting of:
 (1) a variant of adalimumab exhibiting pH-sensitive binding to the TNF antigen,   (2) a variant of adalimumab that is D2E7SS22 and comprises a heavy chain variable region sequence of SEQ ID NO:1 and a light chain variable region sequence of the light chain of SEQ ID NO:2,   (3) a variant of adalimumab comprising a variant Fc region, and   (4) a variant of adalimumab comprising a variant Fc region that 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).   
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . The composition of  claim 1 , wherein the Fc binding polypeptide is a dual variable domain immunoglobulin (DVD-Ig). 
     
     
         17 . The composition of  claim 1 , 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 or a knockdown of one or both the alleles of a Beta galactosidase gene. 
     
     
         19 . (canceled) 
     
     
         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. 
     
     
         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 is selected from the group consisting of
 (1) a second population of Fc domain-containing binding proteins having a G1/2:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1,   (2) a second population of Fc domain-containing binding proteins having a GS:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1, and   (4) a second population of Fc domain-containing binding proteins having a Gtotal:M ratio of at least 10:1, at least 50:1, at least 80:1, or at least 99:1.   
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . The method of  claim 21 , 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 the polypeptide sequence of etanercept, infliximab, adalimumab, or golimumab or the polypeptide sequence of a variant of etanercept, infliximab, adalimumab, or golimumab. 
     
     
         29 . (canceled) 
     
     
         30 . The method of  claim 28 , wherein the variant of adalimumab is selected from the group consisting of
 (1) a variant of adalimumab that exhibits pH-sensitive binding to the TNF antigen,   (2) a variant of adalimumab that is D2E7SS22 and comprises a heavy chain variable region sequence of SEQ ID NO: 1 and a light chain variable region sequence of the light chain of SEQ ID NO:2.   (3) a variant of adalimumab comprising a variant Fc region, and   (2) a variant of adalimumab comprising a variant Fc region that 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).   
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         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. 
     
     
         35 . The method of  claim 34 , wherein the host cell line is a CHO cell line containing a heterologous galactosyltransferase gene or a knockdown of one or both the alleles of a Beta galactosidase gene. 
     
     
         36 . (canceled) 
     
     
         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  claim 1 . 
     
     
         39 . The method of  claim 38 , wherein the disorder is a TNFα associated disorder.