Low acidic species compositions and methods for producing and using the same using displacement chromatography

Abstract

The present invention relates to low acidic species (AR) compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, and methods for producing such low AR compositions using displacement chromatography. Methods for using such compositions to treat a disorder, e.g., a disorder in which TNFα is detrimental, are also provided.

We claim: 
     
         1 . A method for producing a low acidic species composition comprising an antibody, or antigen-binding portion thereof, the method comprising:
 (a) contacting a first sample comprising the antibody, or antigen-binding portion thereof, with a chromatography media, wherein the antibody, or antigen-binding portion thereof, binds to the chromatography media;   (b) displacing the antibody, or antigen-binding portion thereof, bound to the chromatography media with a displacing buffer comprising at least one displacer molecule; and   (c) collecting a chromatography sample,   wherein the chromatography sample comprises a composition of the antibody, or antigen-binding portion thereof, which contains less than about 10% acidic species, thereby producing a low acidic species composition comprising an antibody, or antigen-binding portion thereof.   
     
     
         2 . The method of  claim 1 , wherein the pH of the displacing buffer is lower than the isoelectric point of the antibody, or antigen-binding portion thereof. 
     
     
         3 . The method of  claim 1 , wherein the displacing buffer carries positive charge and wherein the concentration of the displacer in the displacing buffer is at least about 0.1 mM. 
     
     
         4 . The method of  claim 1 , wherein the conductivity of the displacing buffer is about 2 mS/cm to about 20 mS/cm. 
     
     
         5 . The method of  claim 1 , wherein one displacing buffer is used. 
     
     
         6 . The method of  claim 1 , wherein a first displacing buffer and a second displacing buffer are used, and wherein the first and second displacing buffers comprise different concentrations of displacer. 
     
     
         7 . The method of  claim 1 , wherein the method is run in a linear displacement chromatography mode, a two-step displacement chromatography mode, or a multiple-step displacement chromatography mode. 
     
     
         8 . The method of  claim 1 , wherein the chromatography sample comprises a reduced level of host cell proteins as compared to the first sample. 
     
     
         9 . The method of  claim 1 , wherein the chromatography sample comprises a reduced level of one or more of charge variants, structure variants or fragmentation variants as compared to the first sample. 
     
     
         10 . The method of  claim 9 , wherein the chromatography sample comprises a reduced level of basic species as compared to the first sample. 
     
     
         11 . The method of  claim 10 , wherein the reduced level of basic species comprise a reduced level of a lysine species Lys 0 as compared to the first sample. 
     
     
         12 . The method of  claim 1 , wherein the chromatography sample comprises a reduced level of aggregates as compared to the first sample. 
     
     
         13 . The method of  claim 1 , wherein the chromatography sample comprises a reduced level of antibody fragments as compared to the first sample. 
     
     
         14 . The method of  claim 1 , wherein the antibody, or antigen-binding portion thereof, is an anti-TNFα antibody, or antigen-binding portion thereof. 
     
     
         15 . A low acidic species composition comprising an antibody, or antigen-binding portion thereof, wherein the composition comprises less than about 15% acidic species. 
     
     
         16 . The composition of  claim 15 , wherein the acidic species comprise a first acidic region (AR1) and a second acidic region (AR2). 
     
     
         17 . The composition of  claim 15 , wherein the composition comprises about 5% or less acidic species (AR). 
     
     
         18 . The composition of  claim 15 , wherein the composition comprises about 3% or less acidic species (AR). 
     
     
         19 . The composition of  claim 16 , wherein the composition comprises about 0.1% or less AR1 and about 3% or less AR2. 
     
     
         20 . The composition of  claim 17 , wherein the composition comprises about 1.4% or less acidic species (AR). 
     
     
         21 . The composition of  claim 16 , wherein the composition comprises about 1.4% AR2 and about 0.0% AR1. 
     
     
         22 . The composition of  claim 15 , wherein the composition exhibits increased cartilage tissue penetration, increased TNF affinity, reduced cartilage destruction, reduced bone erosion, reduced synovial proliferation, reduced cell infiltration, reduced chondrocyte death, or reduced proteoglycan loss as compared to a non-low acidic species composition, or
 wherein the composition exhibits increased protection against the development of arthritic scores or histopathology scores as compared to a non-low acidic species composition when administered to an animal model of arthritis.   
     
     
         23 . A low acidic species composition comprising an anti-TNFα antibody, or antigen-binding portion thereof, comprising a light chain variable region (LCVR) having a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 7, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3; and a heavy chain variable region (HCVR) having a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 6, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, wherein the composition comprises less than about 10% acidic species (AR). 
     
     
         24 . The composition of  claim 23 , wherein the anti-TNFα antibody, or antigen-binding portion thereof, comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1 and a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2. 
     
     
         25 . The composition of  claim 23 , wherein the anti-TNFα antibody, or antigen-binding portion thereof, is adalimumab, or an antigen binding-portion thereof. 
     
     
         26 . The composition of  claim 23 , wherein the acidic species comprise a first acidic region (AR1) and a second acidic region (AR2), and wherein the composition comprises about 0.1% or less AR1 and about 3% or less AR2. 
     
     
         27 . The composition of  claim 23 , wherein the composition exhibits increased cartilage tissue penetration, increased TNF affinity, reduced cartilage destruction, reduced bone erosion, reduced synovial proliferation, reduced cell infiltration, reduced chondrocyte death, or reduced proteoglycan loss as compared to a non-low acidic species composition, or
 wherein the composition exhibits increased protection against the development of arthritic scores or histopathology scores as compared to a non-low acidic species composition when administered to an animal model of arthritis.   
     
     
         28 . The composition of  claim 23 , wherein the acidic species are product preparation-derived acidic species, cell-culture-derived acidic species or storage-derived acidic species. 
     
     
         29 . A method for treating a subject having a disorder in which TNFα is detrimental, comprising administering to the subject the composition of  claim 15 , thereby treating the subject having a disorder in which TNFα is detrimental. 
     
     
         30 . The method of  claim 29 , wherein the disorder in which TNFα is detrimental is rheumatoid arthritis (RA), psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's Disease, Ulcerative Colitis, juvenile idiopathic arthritis, active axial spondyloarthritis or non-radiographic axial spondyloarthritis.