![]() The high-throughput method developed can rapidly determine the optimal separation conditions for column chromatographic separation. The cation-exchange mixed-mode resins (especially MX-Trp-650M) were more suitable for direct capture of rHSA from crude feedstock. The results showed that no feedstock dilution was needed for Capto MMC and MX-Trp-650M, and separation performance of high dynamic binding capacity at 10% breakthrough (∼50 mg/mL), elution recovery (>95%), and purity (>96%) as well as excellent removal of pigments ( A 350 < 0.080) and HCPs (<581 ppm) could be obtained. Moreover, rHSA separation from Pichia pastoris broth was performed on column under optimal conditions based on the high-throughput screening. The optimal elution pH was determined to be pH 7.0 for SP Sepharose FF, pH 8.0 for MX-Trp-650M and Nuvia cPrime, and pH 9.0 for Capto MMC. The loading conditions at pH 4.0 and dilution rate of 1.0 were suitable for SP Sepharose FF, and no dilution worked for mixed-mode resins. The results indicated that resins with cation-exchange groups had better adsorption performance. Capto MMC is a multimodal cation exchanger with a different selectivity, compared with traditional ion exchangers.Protein purification systems are intended. In this work, four commercialized mixed-mode resins (Capto adhere, Capto MMC, MX-Trp-650M, and Nuvia cPrime) and two ion-exchange resins (Q Sepharose FF and SP Sepharose FF) were used for rHSA separation from Pichia pastoris cell culture broth, and loading and elution conditions were optimized via a high-throughput process using microtiter filter plates. ReadyToProcess Capto MMC columns, 1 L and larger columns for clinical manufacturing that minimize the need for cleaning and cleaning validation.Due to the shortage of blood donation and safe issues of blood-derived pathogens, recombinant human serum albumin (rHSA) expressed by Pichia pastoris is a potential substitute for plasma-derived HSA (pHSA). 7 1.0 M NaOH must only be used for cleaning. 6 Capto MMC can be used under cold-room conditions, but for some proteins the capacity can decrease. 5 pH range where resin can be subjected to cleaning- or sanitization-in-place without significant change in function. Process Characterization Kit Capto MMC, designed for studying the potential impact that resin ligand density might have on the process outcome. 4 pH range where resin can be operated without significant change in function. HiTrap Capto MMC columns, for easy use with a syringe, peristaltic pump, or chromatography system. Lab scale: prepacked for automated purification and analysis Lab scale: prepacked for manual purification HiScreen Capto MMC columns, optimized for method and process development.PreDictor Capto MMC RoboColumn™ units, miniaturized columns for high-throughput process development.PreDictor Capto MMC filter plates, 96-well plates for high-throughput screening or process development.In this work, we evaluated this resin using Cytivas Capto MMC ImpRes, a well-established mixed-mode resin with similar properties, as a benchmark. Prepacked columns and other formatsĬapto MMC resin is available in the following formats: Process development scale: prepacked Diamond MMC Mustang is a relatively new mixed-mode resin, which mediates both cation exchange and hydrophobic interactions. Hydrogen bonding and hydrophobic interactions are also involved. Thus, its features partly resemble a weak cation exchanger. Hydrogen bonding and hydrophobic interactions It can also be used to solve specific purification challenges at both high and low ionic strengths. This resin can be used to bind proteins at the conductivity of the feed material.
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