A fast and efficient purification platform for cell-based influenza viruses by flow-through chromatography.
Since newly emerging influenza viruses with pandemic potentials occurred in recent years, the demand for producing pandemic influenza vaccines for human use is high. For the development of a quick and efficient vaccine production, we proposed an efficient purification platform from the harvest to the purified bulk for the cell-based influenza vaccine production. This platform based on flow-through chromatography and filtration steps and the process only involves a few purification steps, including depth filtration, inactivation by formaldehyde, microfiltration, ultrafiltration, anion-exchange and ligand-core chromatography and sterile filtration. In addition, in the proposed chromatography steps, no virus capture steps were employed, and the purification results were not affected by the virus strain variation, host cells and culturing systems. The results from different virus strains which produced by Vero or MDCK cells in different culturing systems also obtained 33-46% HA recovery yields by this platform. The overall removal rates of the protein and DNA concentration in the purified bulk were over 96.1% and 99.7%, respectively. The low residual cellular DNA concentrations were obtained ranged from 30 to 130pg per human dose (15µg/dose). All influenza H5N1 purified bulks met the regulatory requirements for human vaccine use.
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan; Institute of Molecular and Cellular Biology, National Tsing Hua University, No. 101, Section 2, Kuang Fu Road, Hsinchu 300, Taiwan.
Yu-Fen
YF
Tseng
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan.
Tsai-Chuan
TC
Weng
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan.
Chia-Chun
CC
Lai
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan.
Po-Ling
PL
Chen
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan.
Min-Shi
MS
Lee
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan. Electronic address: alanhu@nhri.org.tw.
Alan Yung-Chih
AY
Hu
D000818:Animals; D002522:Chlorocebus aethiops; D002845:Chromatography / Q000379:methods*; D004285:Dogs; D005374:Filtration; D053124:Influenza A Virus, H5N1 Subtype / Q000302:isolation & purification*; D007252:Influenza Vaccines; D061985:Madin Darby Canine Kidney Cells; D008854:Microscopy, Electron; D014709:Vero Cells
D007252:Influenza Vaccines
Cell culture; Flow-through chromatography; Influenza vaccine; Purification process