Hydrophobic interaction chromatography as polishing step enables obtaining ultra-pure recombinant antibodies

Hydrophobic interaction chromatography is a versatile method to polish antibodies. Here, we present a polishing procedure in order to obtain an ultra-pure preparation of antitumor necrosis factor (TNF) alpha IgG₁. Hydrophobic interaction chromatography (HIC) was used with Toyopearl® Phenyl 650M adsorbent in the presence of ammonium sulfate. Adsorption isotherms, breakthrough curves and chromatographic runs were carried out. The eluted antibody was recovered with 99.9 % purity and 96.2 % yield. In the main peak, aggregates, host cell proteins (HCP) and DNA content were below the limit of detection of the analytical methods used. Thus, the method proposed here shows potential to be employed in a downstream process when an ultra-pure antibody preparation is required.