![]() ![]() ![]() Ionic interactions are commonly involved, although hydrogen bonding and hydrophobic interactions can also be significant. 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. Multimodal ligand The multimodal cation exchanger ligand of Capto MMC ImpRes is immobilized to the base matrix (Fig 2) and interacts with the target molecule through multiple types of 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. The multimodal functionality of Capto MMC ImpRes gives a different selectivity compared with traditional ion exchange columns and it binds proteins at high or low ionic strength. 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. Columns for screening of selectivity, binding, and elution conditions, as well as for small scale purifications. For purifying a WuXiBody-based symmetric bsAb, we identified a LC-missing species as a major byproduct. Fractions were collected and analyzed for aggregate concentration by analytical gel filtration. Sample load was varied between 45 and 80 of dynamic binding capacity (DBC) at residence times of 1RT 2 min or 4 min. BsAbs based on WuXiBody can adopt either asymmetric or symmetric format. 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. Dynamic binding capacity at 10 breakthrough as measured at a residence time of 2 minutes, 300 cm/h in a Tricorn 5/100 column with 10 cm bed height in 50 mM Na-acetate, pH 4.75, 250 mM NaCl. removal capacity of Capto adhere ImpRes resin, a study in B/E mode was performed. 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. WuXiBody is a bispecific antibody (bsAb) platform developed by WuXi Biologics. ![]()
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