Conflicts of interest: none.
Development of a platform process for adenovirus purification that removes human set and nucleolin and provides high purity vector for gene delivery†
Article first published online: 18 OCT 2012
Copyright © 2012 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 3, pages 848–856, March 2013
How to Cite
Riske, F., Berard, N., Albee, K., Pan, P., Henderson, M., Adams, K., Godwin, S. and Spear, S. (2013), Development of a platform process for adenovirus purification that removes human set and nucleolin and provides high purity vector for gene delivery. Biotechnol. Bioeng., 110: 848–856. doi: 10.1002/bit.24742
- Issue published online: 18 JAN 2013
- Article first published online: 18 OCT 2012
- Accepted manuscript online: 5 OCT 2012 07:28AM EST
- Manuscript Accepted: 26 SEP 2012
- Manuscript Revised: 23 AUG 2012
- Manuscript Received: 17 MAY 2012
- gene therapy;
- purification process;
The manufacturing of virus occurs at a modest scale in comparison to many therapeutic proteins mainly because a gene therapy dose is typically only µg of vector. Although modest in scale the generation of high purity virus is challenging due to low viral expression levels and the difficulties in adequately characterizing such a large and complex molecule. A 100 L bioreactor might produce only 100 mg of virus that must be separated from host and process impurities that are typically greater by several orders of magnitude. Furthermore, in the later purification stages the main milieu component is often virus at low concentration (µg/mL) which may non-specifically adsorb to purification surfaces resulting in a lowered virus recovery. This study describes our approach to develop a scalable, manufacturable robust process for an Adenovirus (Ad) gene therapy vector. A number of analytical tools were developed to guide the purification design. During process development, two human proteins, SET and nucleolin, were identified in viral preparations. To our knowledge, this is the first time that SET and nucleolin have been described in Ad. In this report we detail a process for their removal and the robust removal of all process, product and host cell impurities. Biotechnol. Bioeng. 2013; 110: 848–856. © 2012 Wiley Periodicals, Inc.