Protein purification methods to reduce acidic species

  • US 9,346,879 B2
  • Filed: 12/29/2014
  • Issued: 05/24/2016
  • Est. Priority Date: 04/20/2012
  • Status: Active Grant
First Claim
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1. A method for producing a composition comprising adalimumab, the method comprising:

  • contacting a first sample comprising adalimumab comprising more than 10% total acidic species of adalimumab to a first chromatography media in the presence of a loading buffer, wherein the first chromatography media is selected from the group consisting of an ion exchange chromatography media, an affinity chromatography media and a hydrophobic interaction chromatography (HIC) media, andcollecting a first chromatography sample, wherein the first chromatography 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.

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