Chromatographic technology that combined two orthogonal separation modalities exists for viral vectors and vaccines, however not for therapeutic proteins.
Apply the existing purification paradigm from viral vectors and vaccines to proteins.
Single-use cartridge for capturing antibody fragments and host cell proteins from Protein A elutes in flow-through mode
Optimize the mixed-mode ligands and the pore diameter of SEMM-silica beads to achieve efficient capture of the target species and high whole-mAb yield
Optimize particle size of silica beads and cartridge packing to enable fast flow, thus achieving fast processing with minimal pressure drops
Mixed-mode silica resins packed in cartridges for single-use applications, thus eliminating the need for cleaning and sanitization in place
By implementing SEMM‑silica flow‑through polishing (with LigaGuard™ pre‑capture), an organization will reduce downstream polishing operating costs by 30–40% and Protein A resin spend by 50%, enabling a three‑step continuous purification process that increases overall yield by 3–7 percentage points and accelerates throughput by 15–35%.
Demonstrated the applicability of the the paradigm of size-exclusion-mixed-mode chromatographic to the purification of therapeutic proteins Developed a size-exclusion-mixed-mode silica resins (SEMM-silica resins) whose morphology (particle size and pore diameter) and functionalization (ligand identity and chemistry) are tailored for polishing monoclonal antibodies from Protein A eluates Developed operational conditions (buffer composition, concentration, and pH; load ratio; residence time) that achieve >90% product recovery and < 1% antibody fragments/aggregates in the effluent.
Altern, S. H., Kocot, A. J., LeBarre, J. P., Boi, C., Phillips, M. W., Roush, D. J., Menegatti, S., & Cramer, S. M. (2024). Mechanistic model-based characterization of size-exclusion-mixed-mode resins for removal of monoclonal antibody fragments. Journal of Chromatography A, 1718, 464717. https://doi.org/10.1016/j.chroma.2024.464717
LeBarre, J. P., Chu, W., Altern, S. H., Kocot, A. J., Bhandari, D., Barbieri, E., Sly, J., Crapanzano, M., Cramer, S. M., Phillips, M., Roush, D., Carbonell, R., Boi, C., & Menegatti, S. (2024). Mixed-mode size-exclusion silica resin for polishing human antibodies in flow-through mode. Journal of Chromatography A, 1720. https://doi.org/10.1016/j.chroma.2024.464772
Menegatti, S., Presenter, PC3.1-119 Continuous removal of product- and process-related impurities in flow-through mode (PC1.0-035, PC3.1-119, and self-funded research), NIIMBL National Meeting, Washington, D.C., July 27, 2022.
Kocot, A., Altern, S., LeBarre, J., Roush, D., Phillips, M., Menegatti, S., Cramer, S., Model-based Characterization of Size-Exclusion Multimodal Resins. 2022 PREP Symposium, Baltimore, MD, May 17, 2022
LeBarre, J., and Menegatti, S., Removal of antibody fragments from product monomer mAbs in flow-through mode chromatography, ACS National Meeting;,San Diego, CA, March 22, 2022.
Single-use modules for continuous removal of antibody fragments (PC3.1-119), NIIMBL Member Forum, Virtual, June 24, 2021.
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North Carolina State University
Chromagenix
EMD Millipore Corporation
Merck Sharp & Dohme LLC
Rensselaer Polytechnic Institute
