Membrane Protein Expression in Insect Cells Using the Baculovirus Expression Vector System

2020 ◽  
pp. 63-80
Author(s):  
Jacques Boivineau ◽  
Matthias Haffke ◽  
Veli-Pekka Jaakola
Keyword(s):  
Membrane Protein ◽  
Protein Expression ◽  
Expression Vector ◽  
Insect Cells ◽  
Vector System ◽  
Baculovirus Expression Vector System ◽  
Membrane Protein Expression ◽  
Baculovirus Expression ◽  
Baculovirus Expression Vector

The components of shear stress affecting insect cells used with the baculovirus expression vector system

2017 ◽  
Vol 72 (9-10) ◽  
pp. 429-439 ◽  
Author(s):  
Tobias Weidner ◽  
Damir Druzinec ◽  
Martina Mühlmann ◽  
Rainer Buchholz ◽  
Peter Czermak
Keyword(s):  
Surface Area ◽  
Recombinant Proteins ◽  
Expression Vector ◽  
Insect Cells ◽  
Vector System ◽  
Baculovirus Expression Vector System ◽  
Shear Forces ◽  
Baculovirus Expression ◽  
Baculovirus Expression Vector ◽  
Infected Insect

AbstractInsect-based expression platforms such as the baculovirus expression vector system (BEVS) are widely used for the laboratory- and industrial-scale production of recombinant proteins. Thereby, major drawbacks to gain high-quality proteins are the lytic infection cycle and the shear sensitivity of infected insect cells due to turbulence and aeration. Smaller bubbles were formerly assumed to be more harmful than larger ones, but we found that cell damage is also dependent on the concentration of protective agents such as Pluronic®. At the appropriate concentration, Pluronic forms a layer around air bubbles and hinders the attachment of cells, thus limiting the damage. In this context, we used microaeration to vary bubble sizes and confirmed that size is not the most important factor, but the total gas surface area in the reactor is. If the surface area exceeds a certain threshold, the concentration of Pluronic is no longer sufficient for cell protection. To investigate the significance of shear forces, a second study was carried out in which infected insect cells were cultivated in a hollow fiber module to protect them from shear forces. Both model studies revealed important aspects of the design and scale-up of BEVS processes for the production of recombinant proteins.

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