HEK-293F as an alternative cell for the recombinant human granulocyte colony stimulating factor (rhG-CSF) expression

The hG-CSF acts mainly by promoting the granulocytes maturation. It has been used as a biopharmaceutical in the treatment of diseases, particularly neutropenic conditions. A growth curve for HEK-293F cells was constructed and the µmax was 0.0198 and doubling time was 35 hours. Metabolism of HEK-293F cells was analyzed by glutamate/glutamine concentration and an increase in glutamine concentration was observed up to 72 hours (0.214 g/L), followed by a decline until the last day (216 hours). The Geneticin killing curve showed that almost all antibiotic concentration was able to cause cell death of the unmodified cells after 120 hours. Transient transfection with pcDNA3.1_rhG-CSFb vector was done and the recombinant protein was detected after 24 hours. The recombinant protein was identified by immunoblotting and purified by nickel-affinity chromatography. A protein about 20 kDa was detected in SDS-PAGE gel as expected for rhG-CSF. HEK-293F showed to be a good cell for rhG-CSF production.