Downstream process for purification of viral proteins with hydrophobic membrane domain for use in vaccine compositions

A method for purifying a protein from a solution comprising:

• (a) a capture step comprising:
• (i) passing the solution over a first chromatography column, wherein the first chromatography column is a hydrophobic interaction column;
• and
• (ii) eluting a crude protein eluate from the first chromatography column;
• and
• (b) a polish step comprising:
• (i) passing the crude protein eluate obtained from step (a) over a second chromatography column, wherein the second chromatography column is a ligand affinity column;
• (ii) recovering a first flow through intermediate from the second chromatography column;
• (iii) passing the first flow through intermediate obtained from step (b)(ii) over a third chromatography column, wherein the third chromatography column is an anion exchange column;
• (iv) recovering a second flow through intermediate from the third chromatography column;
• (v) passing the second flow through intermediate obtained from step (b)(iv) over a fourth chromatography column, wherein the fourth chromatography column is a ligand affinity column;
• and
• (vi) recovering a purified protein eluate from the fourth chromatography column,
• wherein the purified protein is a hemagglutinin (HA) protein derived from an influenza virus.

A method of purifying a protein from a solution comprising:

• (a) a capture step comprising:
• (i) passing the solution over a first chromatography column, wherein the first chromatography column is a hydrophobic interaction column;
• and
• (ii) eluting a crude protein eluate from the first chromatography column;
• and
• (b) a polish step comprising:
• (i) passing the crude protein eluate obtained from step (a) over a second chromatography column, wherein the second chromatography column is a ligand affinity column;
• (ii) recovering a first flow through intermediate from the second chromatography column;
• (iii) passing the first flow through intermediate obtained from step (b)(ii) over a third chromatography column, wherein the third chromatography column is an anion exchange column;
• (iv) recovering a second flow through intermediate from the third chromatography column;
• (v) passing the second flow through intermediate obtained from step (b)(iv) over a fourth chromatography column, wherein the fourth chromatography column is a ligand affinity column;
• and
• (vi) recovering a purified protein eluate from the fourth chromatography column,
• wherein the purified protein is a fusion (F) protein derived from a respiratory syncytial virus (RSV).