Rosenlöcher, J., Weilandt, C., Sandig, G., Reinke, S.O., Blanchard, V. and Hinderlich, S.
Notes: Human TE671 cells were tested for expression and glycosylation of two proteins, A1AT and GM-CSF. Initial work focused on comparison of TurboFect Transfection Reagent (Life Technologies) and AppliFect LowTox (AppliChem) for expression of A1AT. AppliFect proved superior for expression of the protein and maximum expression achieved through optimization. Both TurboFect and AppliFect were examined for expression of GM-CSF and low transfection efficiencies were obtained despite optimization of conditions and the researchers turned to ViaFect™ Transfection Reagent for improvement. The standard protocol for ViaFect™ Reagent produced results similar to optimized AppliFect but through optimization, the ViaFect™ Reagent were able to produce much higher protein yields (8 × 104 cells; 0.5µg DNA/2µl ViaFect™ Reagent in 24-well plates). Enough protein was produced from transiently transfected cells, scaled to a 10cm dish, to allow development of an N-glycan profile for the recombinant GM-CSF. Interestingly, the authors went back to examine whether ViaFect™ could improve A1AT expression but did not find a difference large enough to change A1AT expression protocols. (4672)