A Fully Continuous Downstream Process for High Titer Products

Prepare a new intensified, continuous purification process for antibodies by eliminating “Protein A” affinity chromatography, by employing precipitation capture purification, tangential flow filtration, and flow-through chromatography for polishing.

Project status

100% Completed

Industry Need

New purification processes are required to meet the global need for high-purity, high-volume, high-dose protein therapeutics.
Monoclonal antibodies (mAbs) to treat pandemic infection, high cholesterol, and Alzheimer’s disease are at the forefront of this need.
“Protein A” affinity chromatography is the bottleneck of the current “platform” mAb manufacturing process.

Solution

The team developed a new type of intensified, continuous purification process that can meet the industry need by eliminating “Protein A” affinity chromatography, the bottleneck of the current “platform” mAb manufacturing process.

This new process employs:

Precipitation for capture purification
Tangential flow filtration for concentration and washing
Flow-through chromatography for polishing

The process can be operated in an integrated, fully continuous mode, increasing volumetric throughput while reducing buffer usage, cost-of-goods, and equipment and supply chain performance bottlenecks of Protein A-based mAb processing that limit global manufacturing capabilities.

Project steps included:

Screening up to 10 different crude mAbs from biomanufacturing partners to determine suitability and conditions for the new process, with up to 5 of the best-suited mAbs purified in the integrated process at bench-scale in extended time demonstration runs.
Transferring the precipitation-based capture technology to integrated pilot-scale equipment for further extended performance testing using 2 mAbs from the bench-scale demonstration.

Outputs/Deliverables

OUTPUTS

Achieved performance metrics that met or exceeded those of the protein A platform process in demonstration runs with three mAbs/mAb-related species
Continuous precipitation-based capture demonstration equipment established at Rensselaer
Parallel project with CESMII enabled instrumentation and smart manufacturing tool (CESMII Smart Manufacturing Innovation Platform) incorporation into the precipitate generation and dewatering operations of the capture step; opportunity for future collaboration
Multiple personnel-in-training at Rensselaer and Penn State worked on project, receiving training in downstream bioprocessing research and development via hands-on experience and abundant interactions with industrial partner personnel
5 graduate research assistant directly supported, 1 unsupported
2 unsupported undergraduate research assistants
1 unsupported postdoctoral researcher
1 paper and 12 conference presentations given in domestic, international venues thus far
Aspects of work incorporated into three courses at Rensselaer and into downstream processing workshop at Penn State

Impacts

Will provide results needed to facilitate widespread industry adoption of this new process.

Will enable de-risking of an innovative recombinant protein purification process that is based on non-traditional continuous precipitation and flowthrough chromatography purification operations

Will offer relief from the capacity, buffer usage, cost, and supply chain bottlenecks of the current "platform" purification process for antibodies

Will reduce process and supply chain complexity rendering process suitable for global deployment, including low-resource environments

Publications

Minervini, M., Mergy, M., Zhu, Y., Gutierrez Diaz, M. A., Pointer, C., Shinkazh, O., Oppenheim, S. F., Cramer, S. M., Przybycien, T. M., & Zydney, A. L. (2023). Continuous precipitation-filtration process for initial capture of a monoclonal antibody product using a four-stage countercurrent hollow fiber membrane washing step. Biotechnology and Bioengineering. https://doi.org/10.1002/bit.28525