Transient Gene Expression in Suspension Hek293 Cells: Application to Large-Scale Protein Production

2005 ◽  
pp. 365-367
Author(s):  
L. Baldi ◽  
R. Jacquet ◽  
S. Picasso ◽  
P. Tromba ◽  
E. Derow ◽  
...  
Keyword(s):  
Gene Expression ◽  
Large Scale ◽  
Protein Production ◽  
Transient Gene Expression ◽  
Hek293 Cells

Rational design of transient gene expression process with lipoplexes for high-level therapeutic protein production in HEK293 cells

2019 ◽  
Vol 85 ◽  
pp. 185-194 ◽  
Author(s):  
Youngji Lee ◽  
Hyeonkyeong Kim ◽  
Eunsil Kim ◽  
Soonyong Park ◽  
Kyoung-Hwa Ryu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rational Design ◽  
Protein Production ◽  
Transient Gene Expression ◽  
Therapeutic Protein ◽  
Hek293 Cells ◽  
High Level

Transient Gene Expression in Suspension HEK-293 Cells: Application to Large-Scale Protein Production

2008 ◽  
Vol 21 (1) ◽  
pp. 148-153 ◽  
Author(s):  
Lucia Baldi ◽  
Natalie Muller ◽  
Sylviane Picasso ◽  
Raymond Jacquet ◽  
Philippe Girard ◽  
...  
Keyword(s):  
Gene Expression ◽  
Large Scale ◽  
Protein Production ◽  
Transient Gene Expression ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

Poly(ethyleneimine)-Mediated Large-Scale Transient Gene Expression: Influence of Molecular Weight, Polydispersity and N -Propionyl Groups

2012 ◽  
Vol 12 (5) ◽  
pp. 628-636 ◽  
Author(s):  
Zuzana Kadlecova ◽  
Sophie Nallet ◽  
David L. Hacker ◽  
Lucia Baldi ◽  
Harm-Anton Klok ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Weight ◽  
Large Scale ◽  
Transient Gene Expression

Large Scale Transient Gene Expression in Mammalian Cells

2006 ◽  
pp. 113-116
Author(s):  
E. -J. Schlaeger ◽  
K. Christensen ◽  
G. Schmid ◽  
N. Schaub ◽  
B. Wipf ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammalian Cells ◽  
Large Scale ◽  
Transient Gene Expression ◽  
Expression In Mammalian Cells

Transient gene expression: Recombinant protein production with suspension-adapted HEK293-EBNA cells

2001 ◽  
Vol 75 (2) ◽  
pp. 197-203 ◽  
Author(s):  
Petra Meissner ◽  
Horst Pick ◽  
Alexandra Kulangara ◽  
Philippe Chatellard ◽  
Kirstin Friedrich ◽  
...  
Keyword(s):  
Gene Expression ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Transient Gene Expression

scholarly journals Rapid, high-yield production of full-length SARS-CoV-2 spike ectodomain by transient gene expression in CHO cells

2020 ◽  
Author(s):  
Matthew Stuible ◽  
Christian Gervais ◽  
Simon Lord-Dufour ◽  
Sylvie Perret ◽  
Denis L’Abbe ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammalian Cells ◽  
Affinity Purification ◽  
Transient Gene Expression ◽  
Full Length ◽  
Spike Protein ◽  
Hek293 Cells ◽  
High Yield ◽  
Diagnostic Tools ◽  
Purification Method

ABSTRACTRecombinant forms of the spike protein of SARS-CoV-2 and related viruses have proven difficult to produce with good yields in mammalian cells. Given the panoply of potential COVID-19 diagnostic tools and therapeutic candidates that require purified spike protein and its importance for ongoing SARS-CoV-2 research, we have explored new approaches for spike production and purification. Three transient gene expression methods based on PEI-mediated transfection of CHO or HEK293 cells in suspension culture in chemically-defined media were compared for rapid production of full-length SARS-CoV-2 ectodomain. A high-cell-density protocol using DXB11-derived CHOBRI/rcTA cells gave substantially better yields than the other methods. Different forms of the spike were expressed, including the wild-type SARS-CoV-2 sequence and a mutated/stabilized form (to favor expression of the full-length spike in prefusion conformation), with and without fusion to putative trimerization domains. An efficient two-step affinity purification method was also developed. Ultimately, we have been able to produce highly homogenous preparations of full-length spike, both monomeric and trimeric, with yields of 100-150 mg/L. The speed and productivity of this method support further development of CHO-based approaches for recombinant spike protein manufacturing.

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