Paper Authors and Title

Mohamed Abdelaal (Atlantic Technological University), Eva Campion, “Optimisation of Electroporation based transfection system in CHO-DP12 suspension cells”


Transfection of DNA plays a major role in production of biopharmaceuticals especially in suspension cells as its easier to upscale the production. An improvement is needed in the process as there is little information about transfection in CHO-DP12 cell line in literature. Here several electroporation experiments were undertaken using the pEZ-M03 plasmid in the CHO-DP12 cell line to optimise transfection efficiency.

Initially plasmid DNA was isolated, and the integrity of the construct was analysed using microvolume spectrophotometer and restriction digestion analysis. A midi-preparation was carried out to obtain higher concentration, quality DNA for transfection. Comparative analysis between PBS and Opti-MEM buffers indicated that PBS buffer promoted a higher transfection efficiency and thus was the most suitable for transfection. Various electroporation using different concentrations of DNA and voltages were undertaken to optimize the transfection.

When the concentration of DNA remained constant (2µg) the transfection efficiency decreased as the voltage increased; the highest transfection efficiency achieved (5%) resulted from the lowest voltage employed (200V). In fact, when the voltage was increased to 230 and 250V no transfection of the cells was recorded. When the concentration of DNA was increased (employing a constant voltage of 190V) the transfection efficiency also increased;3µg of DNA resulted in 14% transfection efficiency and 4 and 5µg resulted in 16% and 22% transfection efficiency respectively.

After further experimentation, if these  results were proved to be repeatable that would have a huge impact on the Biopharma industry as it would lead to improved expression of proteins, faster development of new treatments and at reduced cost.


Download and view Mohamed Abdelaal’s poster.