expression of recombinant factor ix using the transient gene expression technique

Background: pilot and large-scale production of recombinant proteins require the presence of stable clones, but the process of selecting stable clones is time consuming. moreover, continuous clone culturing in large-scale production may cause loss of incoming plasmid and recombinant genes. considering the advancements in transient gene expression (tge) technology, the large-scale expression of factor ix (fix) was investigated in hek cells by the tge technique. materials and methods: hek cells were seeded in a cell factory, and then transfected by pcdna-hfix plasmid using calcium phosphate co-precipitation method. stable hek-hfix cells were also seeded in a cell factory, separately. after adding vitamin k, recombinant fix was quantified in conditioned media using an elisa. moreover, its functional activity was assayed using an aptt test. results: the results showed that the expression and activity of fix by tge technology was, respectively, 1.6 and 1.5 times higher than that obtained through stable hek-fix cells. since calculating the specific activity revealed that for all time periods it is 0.2 mu/ng, so the increase in activity is due to the increase in the amount of fix. conclusions: hek cells with higher transfectability seemed to be an appropriate alternative for transient expression for large-scale protein production. furthermore, if rapid expression of recombinant proteins is intended, tge can replace costly and low-yield methods.