کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8485673 | 1551746 | 2018 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Propagation of Brazilian Zika virus strains in static and suspension cultures using Vero and BHK cells
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موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ایمونولوژی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Propagation of Brazilian Zika virus strains in static and suspension cultures using Vero and BHK cells Propagation of Brazilian Zika virus strains in static and suspension cultures using Vero and BHK cells](/preview/png/8485673.png)
چکیده انگلیسی
Here we present a production platform based on BHK-21 suspension (BHK-21SUS) cells to propagate Brazilian ZIKV at larger quantities in perfusion bioreactors. Scouting experiments performed in tissue culture flasks using adherent BHK-21 and Vero cells have demonstrated similar permissivity and virus yields for four different Brazilian ZIKV isolates. The cell-specific yield of infectious virus particles varied between respective virus strains (1-48Â PFU/cell), and the ZIKV isolate from the Brazilian state Pernambuco (ZIKVPE) showed to be a best performing isolate for both cell lines. However, infection studies of BHK-21SUS cells with ZIKVPE in shake flasks resulted in poor virus replication, with a maximum titer of 8.9Â ÃÂ 103Â PFU/mL. Additional RT-qPCR measurements of intracellular and extracellular viral RNA levels revealed high viral copy numbers within the cell, but poor virus release. Subsequent cultivation in a perfusion bioreactor using an alternating tangential flow filtration system (ATF) under controlled process conditions enabled cell concentrations of about 1.2Â ÃÂ 107Â cells/mL, and virus titers of 3.9Â ÃÂ 107Â PFU/mL. However, while the total number of infectious virus particles was increased, the cell-specific yield (3.3Â PFU/cell) remained lower than determined in adherent cell lines. Nevertheless, the established perfusion process allows to provide large amounts of ZIKV material for research and is a first step towards process development for manufacturing inactivated or live-attenuated ZIKV vaccines.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Vaccine - Volume 36, Issue 22, 24 May 2018, Pages 3140-3145
Journal: Vaccine - Volume 36, Issue 22, 24 May 2018, Pages 3140-3145
نویسندگان
Alexander Nikolay, Leda R. Castilho, Udo Reichl, Yvonne Genzel,