Isolation and Purification of Antibodies Using Protein A Affinity Chromatography

A method for making a pharmaceutical grade preparation comprising adalimumab, wherein the adalimumab is purified from a fermentation harvest of cultured Chinese hamster ovary (CHO) cells expressing adalimumab, and the method comprises:

• a) binding said expressed adalimumab to a Protein A resin at a loading amount of no less than 32 g/L, and
• b) eluting the bound adalimumab at an elution pH of 3.2-3.6 and holding the resulting eluate under acid condition for at least 1 hour, wherein adalimumab monomer level after said holding and prior to any further purification steps is at least 96.1%.

A method for making a pharmaceutical grade preparation comprising adalimumab, wherein said adalimumab is purified from a fermentation harvest of cultured CHO cells expressing adalimumab, and the method comprises:

• a) binding said expressed adalimumab to a Protein A resin, wherein said expressed adalimumab is loaded onto said Protein A resin at a load amount that is 10 to 20% less than the loading amount that results in 10% breakthrough, and
• b) eluting the bound adalimumab at an elution pH of 3.2-3.6 and holding the resulting eluate under acid condition for at least 1 hour, wherein adalimumab monomer level after said holding and prior to any further purification steps is at least 96.1%.