scholarly journals Lentiviral transduction of mammalian cells for fast, scalable and high-level production of soluble and membrane proteins

2018 ◽  
Vol 13 (12) ◽  
pp. 2991-3017 ◽  
Author(s):  
Jonathan Elegheert ◽  
Ester Behiels ◽  
Benjamin Bishop ◽  
Suzanne Scott ◽  
Rachel E. Woolley ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Mammalian Cells ◽  
Lentiviral Transduction ◽  
High Level ◽  
Level Production

scholarly journals High-level production of membrane proteins in E. coli BL21(DE3) by omitting the inducer IPTG

2015 ◽  
Vol 14 (1) ◽  
Author(s):  
Zhe Zhang ◽  
Grietje Kuipers ◽  
Łukasz Niemiec ◽  
Thomas Baumgarten ◽  
Dirk Jan Slotboom ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
E Coli ◽  
High Level ◽  
Level Production

High-level production and purification of biologically active proteins from bacterial and mammalian cells using the tandem pGFLEX expression system

Gene ◽  
1997 ◽  
Vol 193 (2) ◽  
pp. 229-237 ◽  
Author(s):  
Herbert T. Manoharan ◽  
Jean Gallo ◽  
Andrew M. Gulick ◽  
William E. Fahl
Keyword(s):  
Mammalian Cells ◽  
Expression System ◽  
Biologically Active ◽  
High Level ◽  
Level Production

scholarly journals Evaluation of Extracellular Subviral Particles of Dengue Virus Type 2 and Japanese Encephalitis Virus Produced by Spodoptera frugiperda Cells for Use as Vaccine and Diagnostic Antigens

2010 ◽  
Vol 17 (10) ◽  
pp. 1560-1566 ◽  
Author(s):  
Miwa Kuwahara ◽  
Eiji Konishi
Keyword(s):  
Dengue Virus ◽  
Japanese Encephalitis Virus ◽  
Japanese Encephalitis ◽  
Spodoptera Frugiperda ◽  
Mammalian Cells ◽  
Encephalitis Virus ◽  
Commercial Application ◽  
Sf9 Cells ◽  
High Level ◽  
Level Production

ABSTRACT New or improved vaccines against dengue virus types 1 to 4 (DENV1 to DENV4) and Japanese encephalitis virus (JEV), the causative agents of dengue fever and Japanese encephalitis (JE), respectively, are urgently required. The use of noninfectious subviral extracellular particles (EPs) is an inexpensive and safe strategy for the production of protein-based flavivirus vaccines. Although coexpression of premembrane (prM) and envelope (E) proteins has been demonstrated to produce EPs in mammalian cells, low yields have hindered their commercial application. Therefore, we used an insect cell expression system with Spodoptera frugiperda-derived Sf9 cells to investigate high-level production of DENV2 and JEV EPs. Sf9 cells transfected with the prM and E genes of DENV2 or JEV secreted corresponding viral antigens in a particulate form that were biochemically and biophysically equivalent to the authentic antigens obtained from infected C6/36 mosquito cells. Additionally, equivalent neutralizing antibody titers were induced in mice immunized either with EPs produced by transfected Sf9 cells or with EPs produced by transfected mammalian cells, in the context of coimmunization with a DNA vaccine that expresses EPs. Furthermore, the results of an enzyme-linked immunosorbent assay (ELISA) using an EP antigen derived from Sf9 cells correlated significantly with the results obtained by a neutralization test and an ELISA using an EP antigen derived from mammalian cells. Finally, Sf9 cells could produce 10- to 100-fold larger amounts of E antigen than mammalian cells. These results indicate the potential of Sf9 cells for high-level production of flavivirus protein vaccines and diagnostic antigens.

scholarly journals High level production ofEcherichia coliouter membrane proteins OmpA and OmpF intracellularly inBacillus subtilis

1991 ◽  
Vol 83 (1) ◽  
pp. 29-33 ◽  
Author(s):  
Ritvaleena Puohiniemi ◽  
Sarah Butcher ◽  
Eveliina Tarkka ◽  
Matti Sarvas
Keyword(s):  
Membrane Proteins ◽  
High Level ◽  
Level Production

scholarly journals Plant-Based Vaccines: Antigen Design, Diversity, and Strategies for High Level Production

Vaccines ◽  
2022 ◽  
Vol 10 (1) ◽  
pp. 100
Author(s):  
Elizabeth Monreal-Escalante ◽  
Abel Ramos-Vega ◽  
Carlos Angulo ◽  
Bernardo Bañuelos-Hernández
Keyword(s):  
Clinical Trials ◽  
Mammalian Cells ◽  
Vaccine Development ◽  
Antigen Expression ◽  
Microbial Pathogens ◽  
Antigen Production ◽  
Human Use ◽  
High Level ◽  
Design Diversity ◽  
Level Production

Vaccines for human use have conventionally been developed by the production of (1) microbial pathogens in eggs or mammalian cells that are then inactivated, or (2) by the production of pathogen proteins in mammalian and insect cells that are purified for vaccine formulation, as well as, more recently, (3) by using RNA or DNA fragments from pathogens. Another approach for recombinant antigen production in the last three decades has been the use of plants as biofactories. Only have few plant-produced vaccines been evaluated in clinical trials to fight against diseases, of which COVID-19 vaccines are the most recent to be FDA approved. In silico tools have accelerated vaccine design, which, combined with transitory antigen expression in plants, has led to the testing of promising prototypes in pre-clinical and clinical trials. Therefore, this review deals with a description of immunoinformatic tools and plant genetic engineering technologies used for antigen design (virus-like particles (VLP), subunit vaccines, VLP chimeras) and the main strategies for high antigen production levels. These key topics for plant-made vaccine development are discussed and perspectives are provided.

TMCC3 localizes at the three-way junctions for the proper tubular network of the endoplasmic reticulum

2019 ◽  
Vol 476 (21) ◽  
pp. 3241-3260
Author(s):  
Sindhu Wisesa ◽  
Yasunori Yamamoto ◽  
Toshiaki Sakisaka
Keyword(s):  
Endoplasmic Reticulum ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Mammalian Cells ◽  
Coiled Coil ◽  
Transmembrane Domains ◽  
Domain Family ◽  
Coiled Coil Domain ◽  
U2os Cells ◽  
Er Membrane

The tubular network of the endoplasmic reticulum (ER) is formed by connecting ER tubules through three-way junctions. Two classes of the conserved ER membrane proteins, atlastins and lunapark, have been shown to reside at the three-way junctions so far and be involved in the generation and stabilization of the three-way junctions. In this study, we report TMCC3 (transmembrane and coiled-coil domain family 3), a member of the TEX28 family, as another ER membrane protein that resides at the three-way junctions in mammalian cells. When the TEX28 family members were transfected into U2OS cells, TMCC3 specifically localized at the three-way junctions in the peripheral ER. TMCC3 bound to atlastins through the C-terminal transmembrane domains. A TMCC3 mutant lacking the N-terminal coiled-coil domain abolished localization to the three-way junctions, suggesting that TMCC3 localized independently of binding to atlastins. TMCC3 knockdown caused a decrease in the number of three-way junctions and expansion of ER sheets, leading to a reduction of the tubular ER network in U2OS cells. The TMCC3 knockdown phenotype was partially rescued by the overexpression of atlastin-2, suggesting that TMCC3 knockdown would decrease the activity of atlastins. These results indicate that TMCC3 localizes at the three-way junctions for the proper tubular ER network.

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