I use shakers for transient expression with HEK, however the expression level is low and I have to increase the productivity. Will I get more protein in a WAVE? Do you have any other tip for me?


The efficiency of transient transfection is very much dependent on the method used for DNA transfer. Most commonly polyethylenimine (PEI) is used as transfection reagent. The optimum DNA:PEI ratio is dependent on the cultivation medium used and has to be established first. Also the maximum PEI concentration the cells tolerate and the requirement for media exchanges before and possibly after transfection need to be tested. An example procedure can be found in: Fliedl L. et al. 2011 J Biotechnol 153:15-21
Transient transfections have been successfully performed in WAVE Bioreactors e.g. Geisse S et al. 2005 J Struct Funct Genomics 6:165-170. Cell cultivation in a bioreactor may be able to increase the expression level due to the improved culture control and also improved options for feeding of the culture. Additionally larger cultivation volumes are handled more easily in a bioreactor than in multiple parallel shake flasks.

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