A membrane-based purification process for cell culture-derived influenza A virus

作者：Weigel Thomas^*; Solomaier Thomas; Wehmeyer Sebastian; Peuker Alessa; Wolff Michael W; Reichl Udo

来源：Journal of Biotechnology, 2016, 220: 12-20.

DOI：10.1016/j.jbiotec.2015.12.022

• Influenza virus vaccine purification process
• Salt-tolerant membrane chromatography
• Downstream process
• Sulfated cellulose membrane adsorber
• Primary amine anion exchange membrane
• Pseudo-affinity membrane
• chromatography

A simple membrane-based purification process for cell culture-derived influenza virus was established that relies on only two chromatographic unit operations to achieve the contamination limits required according to regulatory authorities. After clarification and concentration, a pseudo-affinity membrane adsorber (sulfated cellulose, SCMA) was applied for virus capture. The subsequent polishing step consisted of a salt-tolerant anion exchange membrane adsorber (STMA) to bind residual DNA. For the presented process neither a buffer exchange step nor a nuclease step for further DNA digestion were required. As a starting point, a two-salt strategy (including a polyvalent ion) was employed to screen STMA conditions in a 96-well plate format. After optimization on chromatographic laboratory scale, the virus recovery was up to 97% with a residual DNA level below 0.82%. In addition, the STMA was characterized regarding its dynamic binding capacity and the impact of flow rate on yields and contamination levels. Overall, the total virus yield for influenza virus A/PR/8/34 (H1/N1) of this two-step membrane process was 75%, while the protein and the DNA contamination level could be reduced to 24% and at least 0.5%, respectively. With 19.8 tig protein and 1.2 ng DNA per monovalent dose, this purity level complies with the limits of the European Pharmacopeia for cell culture-derived vaccines for human use. Overall, the presented downstream process might serve as a generic and economic platform technology for production of cell culture-derived viruses and viral vectors.

• 出版日期2016-2-20