Quantitative analysis of cellular proteome alterations in human influenza A virus-infected mammalian cell lines

PROTEOMICS ◽  
2009 ◽  
Vol 9 (12) ◽  
pp. 3316-3327 ◽  
Author(s):  
Diana Vester ◽  
Erdmann Rapp ◽  
Dörte Gade ◽  
Yvonne Genzel ◽  
Udo Reichl
Keyword(s):  
Quantitative Analysis ◽  
Influenza A Virus ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Influenza A ◽  
Human Influenza ◽  
Mammalian Cell Lines

Harvesting and concentration of human influenza A virus produced in serum-free mammalian cell culture for the production of vaccines

2007 ◽  
Vol 97 (1) ◽  
pp. 73-85 ◽  
Author(s):  
Bernd Kalbfuss ◽  
Yvonne Genzel ◽  
Michael Wolff ◽  
Anke Zimmermann ◽  
Robert Morenweiser ◽  
...  
Keyword(s):  
Cell Culture ◽  
Influenza A Virus ◽  
Mammalian Cell ◽  
Influenza A ◽  
Mammalian Cell Culture ◽  
Human Influenza ◽  
Serum Free

scholarly journals Characterization of Nucleoprotein Extracted from Human Influenza A Virus Cultured in Two Different Cell Lines

2013 ◽  
Vol 7 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Yasin Panahi ◽  
Behrokh Farahmand ◽  
Rasoul Soleimani-Stiar ◽  
Reza Saghiri ◽  
SHima Fattahi ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
Cell Lines ◽  
Influenza A ◽  
Human Influenza

scholarly journals A Y161F Hemagglutinin Substitution Increases Thermostability and Improves Yields of 2009 H1N1 Influenza A Virus in Cells

2017 ◽  
Vol 92 (2) ◽  
Author(s):  
Feng Wen ◽  
Lei Li ◽  
Nan Zhao ◽  
Meng-Jung Chiang ◽  
Hang Xie ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Influenza A ◽  
Vero Cells ◽  
Influenza Viruses ◽  
High Yield ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

ABSTRACTVaccination is the primary strategy for influenza prevention and control. However, egg-based vaccines, the predominant production platform, have several disadvantages, including the emergence of viral antigenic variants that can be induced during egg passage. These limitations have prompted the development of cell-based vaccines, which themselves are not without issue. Most importantly, vaccine seed viruses often do not grow efficiently in mammalian cell lines. Here we aimed to identify novel high-yield signatures for influenza viruses in continuous Madin-Darby canine kidney (MDCK) and Vero cells. Using influenza A(H1N1)pdm09 virus as the testing platform and an integrating error-prone PCR-based mutagenesis strategy, we identified a Y161F mutation in hemagglutinin (HA) that not only enhanced the infectivity of the resultant virus by more than 300-fold but also increased its thermostability without changing its original antigenic properties. The vaccine produced from the Y161F mutant fully protected mice against lethal challenge with wild-type A(H1N1)pdm09. Compared with A(H1N1)pdm09, the Y161F mutant had significantly higher avidity for avian-like and human-like receptor analogs. Of note, the introduction of the Y161F mutation into HA of seasonal H3N2 influenza A virus (IAV) and canine H3N8 IAV also increased yields and thermostability in MDCK cells and chicken embryotic eggs. Thus, residue F161 plays an important role in determining viral growth and thermostability, which could be harnessed to optimize IAV vaccine seed viruses.IMPORTANCEAlthough a promising complement to current egg-based influenza vaccines, cell-based vaccines have one large challenge: high-yield vaccine seeds for production. In this study, we identified a molecular signature, Y161F, in hemagglutinin (HA) that resulted in increased virus growth in Madin-Darby canine kidney and Vero cells, two cell lines commonly used for influenza vaccine manufacturing. This Y161F mutation not only increased HA thermostability but also enhanced its binding affinity for α2,6- and α2,3-linked Neu5Ac. These results suggest that a vaccine strain bearing the Y161F mutation in HA could potentially increase vaccine yields in mammalian cell culture systems.

scholarly journals Structure-guided selection of puromycin N-acetyltransferase mutants with enhanced selection stringency for deriving mammalian cell lines expressing recombinant proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alessandro T. Caputo ◽  
Oliver M. Eder ◽  
Hana Bereznakova ◽  
Heleen Pothuis ◽  
Albert Ardevol ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Recombinant Proteins ◽  
Cultured Cells ◽  
Hek293 Cells ◽  
Reaction Products ◽  
Enzyme Form ◽  
X Ray Crystallography ◽  
Mammalian Cell Lines ◽  
Apparent Loss

AbstractPuromycin and the Streptomyces alboniger-derived puromycin N-acetyltransferase (PAC) enzyme form a commonly used system for selecting stably transfected cultured cells. The crystal structure of PAC has been solved using X-ray crystallography, revealing it to be a member of the GCN5-related N-acetyltransferase (GNAT) family of acetyltransferases. Based on structures in complex with acetyl-CoA or the reaction products CoA and acetylated puromycin, four classes of mutations in and around the catalytic site were designed and tested for activity. Single-residue mutations were identified that displayed a range of enzymatic activities, from complete ablation to enhanced activity relative to wild-type (WT) PAC. Cell pools of stably transfected HEK293 cells derived using two PAC mutants with attenuated activity, Y30F and A142D, were found to secrete up to three-fold higher levels of a soluble, recombinant target protein than corresponding pools derived with the WT enzyme. A third mutant, Y171F, appeared to stabilise the intracellular turnover of PAC, resulting in an apparent loss of selection stringency. Our results indicate that the structure-guided manipulation of PAC function can be utilised to enhance selection stringency for the derivation of mammalian cell lines secreting elevated levels of recombinant proteins.

RAS p21 and other Gn proteins are detected in mammalian cell lines by [γ-35S]GTPγS binding

1989 ◽  
Vol 159 (3) ◽  
pp. 1269-1274 ◽  
Author(s):  
J.G. Comerford ◽  
J.R. Gibson ◽  
A.P. Dawson ◽  
I. Gibson
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines ◽  
Gtpγs Binding ◽  
Ras P21
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