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Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness.

Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness.
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Kim EH, Kwon HI, Park SJ, Kim YI, Si YJ, Lee IW, Kim SM, Kim SI, Ahn DH, Choi YK,


Kim EH, Kwon HI, Park SJ, Kim YI, Si YJ, Lee IW, Kim SM, Kim SI, Ahn DH, Choi YK, (click to view)

Kim EH, Kwon HI, Park SJ, Kim YI, Si YJ, Lee IW, Kim SM, Kim SI, Ahn DH, Choi YK,

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Journal of microbiology and biotechnology 2018 04 12() doi 10.4014/jmb.1712.12007
Abstract

As shown during the 2009 pandemic H1N1 [A(H1N1)pdm09] outbreak, egg-based influenza vaccine production technology is insufficient to meet global demand during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over 10 PFU/ml. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using of a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells (PB1D153N, M1A137T and NS1N176S). In addition, we confirmed that in the context of the high growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than 10PFU/ml) in both attached- and suspended-MDCK cells compared to each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines which are cheap, promptly and vigorously growing which can substitute the egg-based vaccines.Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

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