The data suggest that in comparison with Capto MMC ImpRes, a well-established mixed-mode resin with similar properties, Diamond MMC Mustang exhibits comparable DBC and resolution. In this study we evaluated the performance of Diamond MMC Mustang, a relatively new mixed-mode resin, by conducting intermediate purification of two bsAbs with it. Thus, under the same condition Diamond MMC Mustang exhibits slightly higher DBC than Capto MMC ImpRes for both bsAbs. For comparison purpose, DBC of Capto MMC ImpRes was determined as well, which are 111 and 71 mg/ml for bsAb A and B, respectively. At 5 min residence time, the DBC as determined for bsAb A and B are 123 and 94 mg of protein per ml of resin, respectively. After loading, the column was washed consecutively with 50 mM Tris-HAc, 150 mM NaCl, pH 7.4, 50 mM NaAc-HAc, 1 M NaCl, pH 5.5 and DBC of Diamond MMC MustangĭBC of Diamond MMC Mustang was measured following the procedure described in the method section. The system was run at a flow rate of 180 cm/h (residence time: 5 min). The column was loaded with culture harvest clarified by depth filtration at 30 mg of protein per ml of resin and run in bind-elute mode. The column volume (CV) is approximately 106 ml. MabSelect SuRe LX (Protein A affinity medium) was packed in a 2.6 cm diameter column with 20.0 cm bed height. Precast SurePAGE 4–12% gradient Bis-Tris gels, liquid container and Protein A affinity chromatography Iodoacetamide and SDS were purchased from Sigma-Aldrich (St. Acetic acid and arginine hydrochloride were purchased from J.T.Baker (Phillipsburg, NJ, USA). Thus, this new mixed-mode resin can be a potential substitute of Capto MMC ImpRes for post-Protein A purification.Īmmonium sulfate, disodium hydrogen phosphate dihydrate, ethanol, MES, sodium dihydrogen phosphate dihydrate, sodium acetate trihydrate, sodium chloride, sodium hydroxide and tris(hydroxymethyl)aminomethane were purchased from Merck (Darmstadt, Germany). In addition, MMC Mustang shows good lot-to-lot consistency. The data suggest that in comparison with Capto MMC ImpRes, Diamond MMC Mustang exhibits comparable dynamic binding capacity (DBC) and resolution. In this study we evaluated the performance of Diamond MMC Mustang by conducting intermediate purification of two asymmetric bsAbs with it. As we rely heavily on Capto MMC ImpRes for post capture purification of bsAbs, there is a need for finding a substitute of MMC ImpRes in case the lead time of this resin is significantly extended considering that currently the supply chains of many manufacturers are negatively impacted by the coronavirus pandemic. In fact, Bestchrom's Diamond MMC Mustang and Cytiva's Capto MMC ImpRes share similar characteristics in many respects (Table 1). We previously showed that Capto MMC ImpRes is a powerful and indispensable tool for the purification of recombinant IgG-like bispecific antibody (bsAb), whose production is often associated with various byproducts including half antibody, homodimer and aggregates, ,, ].ĭiamond MMC Mustang is a relatively new mixed-mode resin launched by Bestchrom (Shanghai, China), which also mediates CEX and hydrophobic interactions. The major difference between Capto MMC and Capto MMC ImpRes is that the latter has reduced bead size and ligand density, and hence offers improved resolution. Capto MMC ImpRes from Cytiva (formerly GE Healthcare Life Sciences) is a mixed-mode resin whose ligand is the same as that of Capto MMC, which possesses cation exchange (CEX) and hydrophobic moieties.
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