Purification of recombinantly produced polypeptides
Described herein is a method for separating a recombinantly produced polypeptide from host cell protein. The method includes a step of loading a clarified cell culture supernatant that includes the recombinantly produced polypeptide and the HCP onto a Protein A chromatography column and washing the Protein A chromatography column with a wash buffer comprising a fatty acid having a chain length of at least about 6 carbon atoms, or a fatty acid salt thereof to remove HCP and then recovering the recombinantly produced polypeptide.
1. A method of reducing host cell protein (HCP) level in a composition comprising a recombinantly produced polypeptide, the method comprising:
providing a clarified cell culture supernatant comprising the recombinantly produced polypeptide and one or more HCP;
loading the clarified cell culture supernatant onto a Protein A chromatography column;
washing the Protein A chromatography column with a wash buffer comprising a fatty acid having a chain length of at least about 6 carbon atoms, or a fatty acid salt thereof to remove HCP; wherein the wash buffer comprises between about 25 mM to about 200 mM fatty acid; and wherein the wash buffer comprises sodium chloride present at a concentration of between about 1.0 M to about 2.5 M.
2. The method according to claim 1 , wherein the HCP comprises one or more proteases.
3. The method according to claim 2 , wherein the one or more proteases are selected from: serine proteases, aspartyl proteases, cysteine proteases, metalloproteases, aminopeptidases and combinations thereof.
4. The method of claim 1 , wherein the chain length of the fatty acid or fatty acid salt is between 6 and 12 carbon atoms.
5. The method of claim 1 , wherein the wash buffer has a pH between about 7 to about 9.
6. The method of claim 1 , further comprising a step of clarifying the cell culture harvest to obtain a clarified cell culture harvest and loading the clarified cell culture harvest onto the Protein A chromatography column.
7. The method of claim 1 , further comprising equilibrating the loaded Protein A column with an equilibration buffer prior to washing the column with the wash buffer.
8. The method of claim 7 , wherein the equilibration buffer comprises sodium phosphate.
9. The method of claim 8 , wherein the equilibration buffer comprises between about 10 mM and about 100 mM sodium phosphate.
10. The method of claim 1 , further comprising a second wash step after the column is washed with the fatty acid wash buffer.
11. The method of claim 10 , wherein the buffer used in the second wash step comprises sodium phosphate.