Innovative solutions to overcome current challenges in Purification of Monoclonal antibodies, antibody fragments and novel therapeutic proteins
Antibodies, related fragments and engineered constructs are the fastest growing class of protein therapeutic drugs. While conventional mAbs may be sufficiently purified with traditional Protein A affinity capture, followed by ion exchange steps for impurity removal and final polishing, new non-mAb drug molecules are extremely diversified and may not effectively be purified using this platform. In either case, the use of conventional ion exchange chromatography resins requires a feed stream conductivity adjustment by diafiltration or in-line dilution to achieve a sufficient level of protein Dynamic Binding Capacity (DBC). These unit operations are time, equipment and buffer consuming and contribute to increasing overall purification process costs. Novel solutions exist, using sorbents with differentiated selectivity, able to discriminate target protein from aggregates and impurities based on differences of both isoelectric points and hydrophobicity, while having high DBC and clean-in-place features compatible with industry needs.
Novel Mixed-Mode Cation Exchange chromatography resins offer robust alternatives to conventional cation exchangers for mAbs, antibody fragments and various recombinant proteins purification. In contrast with traditional sulphonate (S) or sulfopropyl (SP) cation exchangers, these mixed-mode sorbents maintain high DBC over a broad range of feed conductivity, allowing direct load from feedstock without UF/DF operations. Such a broad conductivity range also offers more process flexibility, according to the ionic strength conditions used during the previous chromatography step or needed for the following purification step.
Mixed-mode chromatography supports:
- Purification of “non-antibody” recombinant proteins through the unique selectivity, which can be optimized through pH and salt tuning to separate target proteins and closely related contaminants
- Removal of aggregates, HCP and various contaminants in various cases very efficient removal of aggregates, residual DNA, host cell proteins (HCPs) or protein misfolding can be achieved, to comply with regulatory end-product requirements
Pall has developed a flexible and scalable purification platform for conventional monoclonal antibody purification based on the innovative CMM HyperCel™ mixed-mode cation exchange resin and several other high performing chromatography sorbents and membranes:
- Protein A affinity capture with KANEKA KanCapA™ sorbent or the new KANEKA KanCapA 3G
- Intermediate DNA/HCP removal via anion exchange (AEX) chromatography on Mustang® Q membrane in flow through mode
- Final polishing/aggregate removal on CMM HyperCel mixed-mode cation exchange resin