375810 Endonuclease Use and Clearance from a Cell Culture-Based Vaccine Process Using TFF

Wednesday, November 19, 2014
Galleria Exhibit Hall (Hilton Atlanta)
Adam Sokolnicki, Biomanufacturing Sciences Network, EMD Millipore Corp., Billerica, MA

Benzonase® endonuclease is a powerful tool for degrading all forms of (deoxy)ribonucleic acids in cell culture harvests to base pair (BP) lengths under 10 units.  It is effective over a wide range of conditions including temperature, pH, and concentrations of Mg2+, detergents, chelating agents, and monovalent cations.  It is often employed in the production of viral vaccines, completely digesting DNA and RNA to improve clearance and reduce solution viscosity.  Removing Benzonase, a 60kD dimer, from the process stream post treatment may be achieved with TFF with the appropriate molecular weight cutoff (MWCO) membrane, typically 300 kD.  This process has heretofore not been well described in the literature, so the present study fills this gap pairing Benzonase treatment of a DNA-spiked inactivated flu virus cell culture harvest with TFF technology, for clearance of the digested DNA and remaining Benzonase by diafiltration.

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See more of this Session: Poster Session: Bioengineering
See more of this Group/Topical: Food, Pharmaceutical & Bioengineering Division