scholarly journals Interferon Inhibition Enhances the Pilot-Scale Production of Rabies Virus in Human Diploid MRC-5 Cells

Viruses ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 49
Author(s):  
Xiao Yang ◽  
Mingming Wan ◽  
Linjun Cai ◽  
Ali Hou ◽  
Bo Sun ◽  
...  

Inactivated vaccines based on cell culture are very useful in the prevention and control of many diseases. The most popular strategy for the production of inactivated vaccines is based on monkey-derived Vero cells, which results in high productivity of the virus but has a certain carcinogenic risk due to non-human DNA contamination. Since human diploid cells, such as MRC-5 cells, can produce a safer vaccine, efforts to develop a strategy for inactivated vaccine production using these cells have been investigated using MRC-5 cells. However, most viruses do not replicate efficiently in MRC-5 cells. In this study, we found that rabies virus (RABV) infection activated a robust interferon (IFN)-β response in MRC-5 cells but almost none in Vero cells, suggesting that the IFN response could be a key limiting factor for virus production. Treatment of the MRC-5 cells with IFN inhibitors increased RABV titers by 10-fold. Additionally, the RABV titer yield was improved five-fold when using IFN receptor 1 (IFNAR1) antibodies. As such, we established a stable IFNAR1-deficient MRC-5 cell line (MRC-5IFNAR1−), which increased RABV production by 6.5-fold compared to normal MRC-5 cells. Furthermore, in a pilot-scale production in 1500 square centimeter spinner flasks, utilization of the MRC-5IFNAR1− cell line or the addition of IFN inhibitors to MRC cells increased RABV production by 10-fold or four-fold, respectively. Thus, we successfully established a human diploid cell-based pilot scale virus production platform via inhibition of IFN response for rabies vaccines, which could also be used for other inactivated virus vaccine production.

Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 290
Author(s):  
Chih-Yu Cheng ◽  
Chia-Huang Tsai ◽  
Pei-Jyun Liou ◽  
Chi-Hang Wang

For pilot-scale production of chito-oligosaccharides, it must be cost-effective to prepare designable recombinant chitosanase. Herein, an efficient method for preparing recombinant Bacillus chitosanase from Escherichia coli by elimination of undesirable substances as a precipitate is proposed. After an optimized culture with IPTG (Isopropyl β-d-1-thiogalactopyranoside) induction, the harvested cells were resuspended, disrupted by sonication, divided by selective precipitation, and stored using the same solution conditions. Several factors involved in these procedures, including ion types, ionic concentration, pH, and bacterial cell density, were examined. The optimal conditions were inferred to be pH = 4.5, 300 mM sodium dihydrogen phosphate, and cell density below 1011 cells/mL. Finally, recombinant chitosanase was purified to >70% homogeneity with an activity recovery and enzyme yield of 90% and 106 mg/L, respectively. When 10 L of 5% chitosan was hydrolyzed with 2500 units of chitosanase at ambient temperature for 72 h, hydrolyzed products having molar masses of 833 ± 222 g/mol with multiple degrees of polymerization (chito-dimer to tetramer) were obtained. This work provided an economical and eco-friendly preparation of recombinant chitosanase to scale up the hydrolysis of chitosan towards tailored oligosaccharides in the near future.


2015 ◽  
Vol 173 ◽  
pp. 827-837 ◽  
Author(s):  
Domien De Paepe ◽  
Katleen Coudijzer ◽  
Bart Noten ◽  
Dirk Valkenborg ◽  
Kelly Servaes ◽  
...  

2017 ◽  
Vol 109 ◽  
pp. 634-645 ◽  
Author(s):  
Svitlana Nitièma-Yefanova ◽  
Valérie Tschamber ◽  
Romain Richard ◽  
Sophie Thiebaud-Roux ◽  
Brice Bouyssiere ◽  
...  

2008 ◽  
Vol 13 (2) ◽  
pp. 182-188 ◽  
Author(s):  
Kang-Ik Jo ◽  
You-Jung Lee ◽  
Bo-Kyung Kim ◽  
Bo-Hwa Lee ◽  
Chung-Han Chung ◽  
...  

2012 ◽  
Vol 24 (6) ◽  
pp. 1353-1360 ◽  
Author(s):  
Jazmin Bazaes ◽  
Claudia Sepulveda ◽  
F. Gabriel Acién ◽  
Juan Morales ◽  
Leonel Gonzales ◽  
...  

2018 ◽  
Vol 08 (01) ◽  
Author(s):  
Milos Beran ◽  
Josef Drahorad ◽  
Ondrej Vltavsky ◽  
Marian Urban ◽  
Iva Laknerova ◽  
...  

1997 ◽  
Vol 32 (3) ◽  
pp. 191-194
Author(s):  
V.A. Samoylenko ◽  
T.V. Yakshina ◽  
R.R. Oleynikov ◽  
Yu.G. Kostenko ◽  
D.N. Spicyna ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document