文献类型：期刊文献

英文题名：Highly efficient production of an influenza H9N2 vaccine using MDCK suspension cells

作者：Wu, Yixiao[1];Jia, Hanjing[1];Lai, Hanzhang[2,3];Liu, Xuping[1,4];Tan, Wen-Song[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd,POB 309, Shanghai 200237, Peoples R China;[2]Guangdong Wens Dahuanong Biotechnol Co Ltd, Key Lab Biotechnol & Bioprod Dev Anim Epidem Prev, Minist Agr, 6 Dongdi North Rd, Yunfu 527400, Xinxing County, Peoples R China;[3]Zhaoqing Dahuanong Biol Med Co Ltd, Guangdong Enterprise Key Lab Biotechnol R&D Vet B, Zhaoqing High Tech Dev Zone, Zhaoqing 526238, Guangdong, Peoples R China;[4]Shanghai BioEngine Sci Tech Co Ltd, 781 Cailun Rd, Shanghai 201203, Peoples R China

年份：2020

卷号：7

期号：1

外文期刊名：BIORESOURCES AND BIOPROCESSING

收录：;EI(收录号：20224513084343);WOS:【SCI-EXPANDED(收录号:WOS:000595373800001)】；

基金：This work was financially supported by the Fundamental Research Funds for the Central Universities (No. 22221818014) and the Development Funds of Shanghai Zhangjiang National Independent Innovation Demonstration Zone (No. ZJ2015-ZD-002).

语种：英文

外文关键词：MDCK suspension cells; Cell adaptation; Influenza virus; Feed strategy; Cell culture-based vaccine manufacturing

摘要：The use of H9N2 subtype avian influenza vaccines is an effective approach for the control of the virus spread among the poultry, and for the upgrading of vaccine manufacturing, cell culture-based production platform could overcome the limitations of conventional egg-based platform and alternate it. The development of serum-free suspension cell culture could allow even higher virus productivity, where a suspension cell line with good performance and proper culture strategies are required. In this work, an adherent Mardin-Darby canine kidney (MDCK) cell line was adapted to suspension growth to cell concentration up to 12 x 10(6) cells/mL in a serum-free medium in batch cultures. Subsequently, the H9N2 influenza virus propagation in this MDCK cell line was evaluated with the optimization of infection conditions in terms of MOI and cell concentration for infection. Furthermore, various feed strategies were tested in the infection phase for improved virus titer and a maximum hemagglutinin titer of 13 log(2) (HAU/50 mu L) was obtained using the 1:2 medium dilution strategy. The evaluation of MDCK cell growth and H9N2 virus production in bioreactors with optimized operating conditions showed comparable cell performance and virus yield compared to shake flasks, with a high cell-specific virus yield above 13,000 virions/cell. With the purified H9N2 virus harvested from the bioreactors, the MDCK cell-derived vaccine was able to induce high titers of neutralizing antibodies in chickens. Overall, the results demonstrate the promising application of the highly efficient MDCK cell-based production platform for the avian influenza vaccine manufacturing.