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

A method for producing a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, the method comprising:

• (a) contacting a first sample comprising the immunoglobulin, wherein the sample comprises more than 10% total acidic species of the immunoglobulin, to an affinity chromatography media in a first loading buffer, and eluting a first eluted sample from the affinity chromatography media;
• (b) contacting the first eluted sample to a second chromatography media in a second loading buffer and eluting a second eluted sample from the second chromatography media;
• and
• (c) contacting the second eluted sample to a third chromatography media in a third loading buffer and eluting a third eluted sample from the third chromatography media;
• wherein the third eluted sample comprises a composition of the immunoglobulin comprising less than 10% total acidic species of the immunoglobulin, wherein the acidic species of the immunoglobulin correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of the immunoglobulin, wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.

A method for producing a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, the method comprising:

• (a) contacting a first sample comprising the immunoglobulin, wherein the sample comprises more than 10% total acidic species of the immunoglobulin, to a protein A chromatography media in a first loading buffer, and eluting a first eluted sample from the protein A chromatography media;
• (b) contacting the first eluted sample to an anion exchange (AEX) chromatography media in a second loading buffer and eluting a second eluted sample from the AEX chromatography media;
• and
• (c) contacting the second eluted sample to a cation exchange (CEX) chromatography media in a third loading buffer and eluting a third eluted sample from the CEX chromatography media;
• wherein the third eluted sample comprises a composition of the immunoglobulin comprising less than 10% total acidic species of the immunoglobulin, wherein the acidic species of the immunoglobulin correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of the immunoglobulin, wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.

A method for producing a composition comprising adalimumab, the method comprising:

• (a) contacting a first sample comprising adalimumab, wherein the sample comprises more than 10% total acidic species of adalimumab, to a protein A chromatography media in a first loading buffer, and eluting a first eluted sample from the protein A chromatography media;
• (b) contacting the first eluted sample to an anion exchange (AEX) chromatography media in a second loading buffer and eluting a second eluted sample from the AEX chromatography media;
• and
• (c) contacting the second eluted sample to a cation exchange (CEX) chromatography media in a third loading buffer and eluting a third eluted sample from the CEX chromatography media;
• wherein the third eluted sample comprises a composition of adalimumab comprising less than 10% total acidic species of adalimumab, wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab, wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.