Continuous Countercurrent Tangential Chromatography for Monoclonal Antibody Purification

Abstract

Recent increases in antibody titers have generated renewed interest in lower-cost continuous chromatographic processes for antibody purification. The objective of this study was to design, build, and test a continuous countercurrent tangential chromatography system in which the resin (in the form of a slurry) flows sequentially through a series of static mixers and hollow fiber membrane modules to accomplish the binding, washing, elution, stripping, and equilibration steps in continuous operation with true countercurrent staging. Experimental studies were performed for the purification of an IgG₄ monoclonal antibody from a model feed containing bovine serum albumin and myoglobin using a protein A affinity resin. Batch uptake / desorption experiments were used in combination with critical flux filtration data to design the continuous countercurrent tangential chromatography system. The process was stable during continuous operation, providing the IgG₄ at 94% yield with greater than 97% purity. The antibody productivity was more than 40 g protein per liter of resin per hour, which is significantly greater than can be obtained using conventional packed columns. The results clearly demonstrate the potential of using continuous countercurrent tangential chromatography for large-scale antibody purification.

Keywords:

• chromatography
• downstream processing
• hollow fiber membrane
• monoclonal antibody
• protein A
• protein purification

ACKNOWLEDGEMENTS

The authors would like to thank Spectrum Laboratories for donation of the MidiKros® hollow fiber membrane modules, Regeneron for donation of the IgG₄, and Life Technologies for donating and discounting the POROS MabCapture A resin. Funding for this project was provided by NIH Phase I SBIR from the National Institute of General Medical Sciences (GRANT10756648) with additional support from the Ben Franklin Technology Partners of Central PA. We would also like to thank our interns Matthew Long and David Currie for providing assistance with some of the experimental studies.