Maximizing productivity of chromatography steps for purification of monoclonal antibodies

✍ Scribed by Nihal Tugcu; David J. Roush; Kent E. Göklen

• Publisher: John Wiley and Sons
• Year: 2007
• Tongue: English
• Weight: 277 KB
• Volume: 99
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.21604

✦ Synopsis

Abstract

The large scale production of monoclonal antibodies presents a challenge to design efficient and cost effective downstream purification processes. We explored a two stage resin screening approach to identify the best candidates to be utilized for the platform purification of monoclonal antibodies. The study focused on commercially available affinity resins including Protein A, mimetic and mixed‐mode interaction resins as well as ion exchangers used in polishing steps. An initial screening using pure proteins was followed by a final screening where selected resins were utilized for the purification of MAbs in complex mixtures. Initial screenings aimed to measure the theoretical upper limit for dynamic binding capacity (DBC) at 1% breakthrough and productivity. We confirmed that DBC of affinity, mimetic and mixed‐mode resins was a strong function of the linear velocity used for loading. Productivities >27 g/(L‐h), were obtained for rProtein A FF, Mabselect and Prosep rA Ultra at 2 min residence time. For the cation exchangers, we identified UNOsphere S and Fractogel SO~3~ as the best candidates for our purification based on DBC. For anion exchangers operated in flowthrough mode, Q Sepharose XL and UNOsphere Q were selected from the initial screening based on DBC and resolution of IgG from BSA. Finally, a three step purification scheme was implemented using the selected affinity and ion exchangers for the purification of IgG from complex feedstocks. We found that Mabselect followed by UNOsphere Q and UNOsphere S provided the best purification scheme for our applications based on productivity. Biotechnol. Bioeng. 2008;99: 599–613. © 2007 Wiley Periodicals, Inc.