Comparison of recombinant protein expression in a baculovirus system in insect cells (Sf9) and silkworm

2010 ◽  
Vol 149 (2) ◽  
pp. 219-227 ◽  
Author(s):  
A. Usami ◽  
S. Ishiyama ◽  
C. Enomoto ◽  
H. Okazaki ◽  
K. Higuchi ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Insect Cells ◽  
Recombinant Protein Expression ◽  
Baculovirus System

scholarly journals Identification of Essential Genetic Baculoviral Elements for Recombinant Protein Expression by Transactivation in Sf21 Insect Cells

PLoS ONE ◽  
2016 ◽  
Vol 11 (3) ◽  
pp. e0149424 ◽  
Author(s):  
Maren Bleckmann ◽  
Margitta Schürig ◽  
Fang-Fang Chen ◽  
Zen-Zen Yen ◽  
Nils Lindemann ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Insect Cells ◽  
Recombinant Protein Expression

scholarly journals Process Optimization for Recombinant Protein Expression in Insect Cells

10.5772/67849 ◽  
2017 ◽  
Author(s):  
Jan Zitzmann ◽  
Gundula Sprick ◽  
Tobias Weidner ◽  
Christine Schreiber ◽  
Peter Czermak
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Process Optimization ◽  
Insect Cells ◽  
Recombinant Protein Expression

scholarly journals Relocation of the attTn7 Transgene Insertion Site in Bacmid DNA Enhances Baculovirus Genome Stability and Recombinant Protein Expression in Insect Cells

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1448
Author(s):  
Gorben P. Pijlman ◽  
Carissa Grose ◽  
Tessy A. H. Hick ◽  
Herman E. Breukink ◽  
Robin van den Braak ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Genome Engineering ◽  
Insect Cells ◽  
Recombinant Protein Expression ◽  
Insertion Site ◽  
Scale Up ◽  
Expression Vectors ◽  
Expression Levels ◽  
Transgene Insertion

Baculovirus expression vectors are successfully used for the commercial production of complex (glyco)proteins in eukaryotic cells. The genome engineering of single-copy baculovirus infectious clones (bacmids) in E. coli has been valuable in the study of baculovirus biology, but bacmids are not yet widely applied as expression vectors. An important limitation of first-generation bacmids for large-scale protein production is the rapid loss of gene of interest (GOI) expression. The instability is caused by the mini-F replicon in the bacmid backbone, which is non-essential for baculovirus replication in insect cells, and carries the adjacent GOI in between attTn7 transposition sites. In this paper, we test the hypothesis that relocation of the attTn7 transgene insertion site away from the mini-F replicon prevents deletion of the GOI, thereby resulting in higher and prolonged recombinant protein expression levels. We applied lambda red genome engineering combined with SacB counterselection to generate a series of bacmids with relocated attTn7 sites and tested their performance by comparing the relative expression levels of different GOIs. We conclude that GOI expression from the odv-e56 (pif-5) locus results in higher overall expression levels and is more stable over serial passages compared to the original bacmid. Finally, we evaluated this improved next-generation bacmid during a bioreactor scale-up of Sf9 insect cells in suspension to produce enveloped chikungunya virus-like particles as a model vaccine.

Sign in / Sign up

Export Citation Format

Share Document