In this study, we demonstrated that when Capto MMC ImpRes chromatography is conducted under weak partitioning mode, high throughput, good yield, and effective byproduct removal are simultaneously achieved.īispecific antibody (bsAb) Capto MMC ImpRes Light chain (LC)-missing byproduct Mixed-mode chromatography Weak partitioning mode WuXiBody.Ĭopyright © 2021 Elsevier Inc. When using Capto MMC ImpRes, protein binding can be expected at pH values higher than those used with traditional ion exchangers due to the contribution from non-electrostatic interactions. However, the dynamic binding capacity (DBC) of Capto MMC ImpRes is relatively low under the selected condition, making the process less desirable for large-scale manufacturing. We previously showed that Capto MMC ImpRes mixed-mode chromatography under bind-elute mode can effectively remove this LC-missing species. While for bsAbs based on other platforms removal of such byproduct can pose considerable challenge to the downstream team, in this case WuXiBody's unique design makes separation relatively straightforward. The information provided in this study allows users to have additional options when selecting mixed-mode resin for intermediate. In addition, the resin under evaluation shows good lot-to-lot consistency. For purifying a WuXiBody-based symmetric bsAb, we identified a LC-missing species as a major byproduct. The data suggest that in comparison with Capto MMC ImpRes, Diamond MMC Mustang exhibits comparable binding capacity and resolution. Both chromatography media extend the well established Capto platform to include high-resolution media. BsAbs based on WuXiBody can adopt either asymmetric or symmetric format. Capto Phenyl ImpRes and Capto Butyl ImpRes are hydrophobic interaction chromatography (HIC) media (resins) developed for the intermediate and polishing steps in a downstream protein purification process (Fig 1). Its key feature is the replacement of one parental antibody's CH1/CL region with the T cell receptor (TCR) constant domain, a design that promotes cognate heavy chain (HC)-light chain (LC) pairing. Capto MMC/Capto MMC ImpRes: high ionic strength solution (e.g., 2 M NaCl in buffer) and at the same time increase pH to > 9 (e.g., Tris buffer with 2 M NaCl) Regenerate the medium by washing until the column effluent shows stable conductivity and pH values. In addition, Capto MMC ImpRes and Capto adhere ImpRes have also both been proposed to be effective polishing steps 26, 78, with Capto MMC ImpRes reported to be able to remove bsAb products such as holehole homodimer and half antibodies through the incorporation of additional pH and NaCl wash steps this, however, occurs at a compromise of. WuXiBody is a bispecific antibody (bsAb) platform developed by WuXi Biologics.
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