Countercurrent Staged Diafiltration for Monoclonal Antibody Formulation
Open Access
- Author:
- Nambiar, Anirudh Manoj Kumar
- Area of Honors:
- Chemical Engineering
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Andrew Zydney, Thesis Supervisor
Andrew Zydney, Thesis Honors Advisor
Dr. Manish Kumar, Faculty Reader - Keywords:
- Ultrafiltration
Diafiltration
Protein
Continuous Processing - Abstract:
- There is significant interest in moving current biomanufacturing operations from traditional batch to continuous processes due to the potential for increased productivity, enhanced product quality, and greater flexibility. Considerable progress has been made in the development of continuous perfusion bioreactors and continuous chromatographic processes. However, the ultrafiltration / diafiltration step, which is currently used for concentration and final formulation of essentially all biotherapeutics, is currently an inherently batch operation. The objective of this project was to examine the feasibility and performance characteristics of a countercurrent staged diafiltration system for continuous protein formulation. Experimental studies were performed using Pall Cadence™ Inline Concentrators, which are specifically designed for continuous single-pass operation with high conversion (ratio of permeate to feed flow rate greater than 50%). Serum IgG was used as a model protein, with vitamin B12 as a model impurity. Data obtained in a 2-stage system with 90% conversion in each stage provided 98.8% removal of vitamin B12 with essentially 100% recovery of the IgG. Impurity removal was increased to 99.7% by increasing the conversion to 95%. The experimental results were in excellent agreement with model calculations based on the countercurrent staged configuration. 99.99% impurity removal could be achieved with 4 countercurrent stages, while using 40% less buffer due to the internal recycling of the buffer between the stages. These results provide the first demonstration that one can successfully use countercurrent staged diafiltration with single-pass tangential flow filtration modules for the continuous formulation of high value biological products.