5556
Abdalla Adel Sabry Abdelrazik Mohamed Elshereef
Optimization of transient protein production by chemically
transfected CHO suspension cells
PEI-transient transfection, GFP-transfection efficiency, viability, TGF-β1 expression.
In the present work, cell viability and transfection efficiency are the most important response parameters evaluated to study such fundamental processes. Transfection efficiency has been investigated by transfecting a plasmid-expressing green fluorescent protein (GFP) into the CHO-K1 cell which can easily be detected and quantitatively measured. CHOMACS CD medium supports cultivation of a high transfected cell density of about 10-18 million cells per mL with a viability of over 80%, which elucidates a transfection culture volume of up to 12.5 mL in a Tubespin®50 bioreactor and in small shake flasks. Several conditions for transfection efficiency (TE) have been explored and guided by a design of experiment approach (DoE). A high TE of 70% combined with a high cell viability of over 95%, and an extended expression time of 120 hours post-transfection, (hpt) have been achieved through a modified repeated transient transfection strategy utilizing a low amount of 6 μg pDNA:30 μg PEI for 10 million cells per mL.
2019
Ph.d
Hanovar
Natureal Science