Methods of cleaning chromatography matrices
AssigneeEli Lilly and Company
InventorsNoah TAN SHIZHAO, Jean LORDAN
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Abstract
Provided herein are improved methods of cleaning a protein A chromatography matrix that has previously been used to purify an Fc-containing protein. The methods generally involve washing the protein A chromatography matrix with a series of three solutions, the first containing an acid, the second containing Tris, and the third containing NaOH (e.g., 0.001-0.075 M NaOH).
Claims46
A method for cleaning a protein A chromatography matrix that has previously been used to purify dulaglutide, the method comprising sequentially washing a chromatography column comprising the protein A chromatography matrix with:
- a) a first solution comprising one or more acid selected from the group consisting of acetic acid and phosphoric acid;
- b) a second solution comprising Tris;
- and
- c) a third solution comprising 0.001-0.075 M NaOH.
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A method for cleaning a protein A chromatography matrix that has previously been used to purify dulaglutide, the method comprising sequentially washing a chromatography column comprising the protein A chromatography matrix with:
- a) 2-3 column volumes of a first solution comprising about 1% acetic acid and about 1% phosphoric acid at a flow rate of about 270 cm/hr;
- b) 1-2 column volumes of a second solution comprising about 50 mM Tris at a pH of about 8 at a flow rate of about 270 cm/hr;
- and
- c) 2-3 column volumes of a third solution comprising about 0.01 M NaOH at a flow rate of about 270 cm/hr;
- wherein the chromatography column has a diameter of about 100 cm and an average particle size of about 85 μm;
- wherein the protein A chromatography matrix a comprises a protein A ligand that has increased stability under alkali conditions relative to wild type Staphylococcus aureus protein A;
- and
- wherein the method does not include a static hold of any of the first solution, the second solution or the third solution.
Dependencies are inferred from the claim text.