A flow-through chromatography purification process for Vero cell-derived influenza virus (H7N9)

• Published in: Journal of virological methods Vol. 301; p. 114408
• Main Authors: Fei, ChengRui, Gao, JingXia, Fei, ChengHua, Ma, Lei, Zhu, WenYong, He, LingYu, Wu, YaNan, Song, ShaoHui, Li, WeiDong, Zhou, Jian, Liao, GuoYang
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2022
• Subjects: Flow-through chromatography; H7N9 influenza virus; Purification process; Vero cell; H7N9 influenza virus; Purification process; Vero cell; Flow-through chromatography
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2021.114408

•The virus strain used in this study was reassorted by reverse genetics.•The virus was cultured with serum-free virus maintenance solution that reduced the difficulty of virus purification in the downstream process.•The flow-through chromatography process avoids the virus capture step and can be used for other influenza virus.•the flow-through process has the advantages of high efficiency, easy operation, fast production in a short period and no restriction by strain. Immunization is the most effective way to respond to an influenza epidemic. To produce Vero cell-derived influenza vaccines, a more efficient, stable and economical purification process is required. In this study, we purified the H7N9 influenza virus grown in Vero cells that were cultured in a serum-free medium by using a combination of anion exchange chromatography (AEC) and ligand-activated core chromatography (LCC), which avoids the virus capture step. After purification, 99.95 % host cell DNA (hcDNA) (final concentration: 28.69 pg/dose) and 98.87 % host cell protein (HCP) (final concentration: 28.28 ng/dose) were removed. The albumin content was 11.36 ng/dose. All these remnants met the current Chinese Pharmacopoeia and WHO requirements. The final virus recovery rate was 58.74 %, with the concentration of hemagglutinin recorded at 132.12 μg/mL. The flow-through chromatography purification process represents an alternative to the existing processes for cell-derived influenza viruses and might be suitable for the purification of other viruses as well.