Single-use modules for continuous removal of antibody fragments

Develop a single-use cartridge packed with a silica-based size-exclusion and mixed-mode (SEMM) adsorbent for capturing monoclonal antibody (mAb) fragments and residual host cell proteins (HCPs) from Protein A-eluates in flow-through mode

Project status

100% Completed

Industry Need

Chromatographic technology that combined two orthogonal separation modalities exists for viral vectors and vaccines, however not for therapeutic proteins.

Solution

Apply the existing purification paradigm from viral vectors and vaccines to proteins.

Outputs/Deliverables

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.

Impacts

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

Publications

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

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