High IFN-α2b productivity obtained from lentivirus-transduced CHO cells

GUGLIOTTA, AGUSTINA; CEAGLIO, NATALIA; PRIETO, CLAUDIO; ETCHEVERRIGARAY, MARINA; KRATJE, RICARDO; OGGERO, MARCOS

Potrero de los Funes

Congreso; XLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2011

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Interferons are cytokines that can interfere viral activity, inhibit cell proliferation and modulate the immune system. Human IFN-alpha2 produced in bacteria is employed in the treatment of cancer and chronic viral diseases. Due to its short half-life, the use of high and repeated doses is necessary, leading to several side effects. In our lab, we have obtained a hyperglycosylated rhIFN-alpha2b mutein (IFN-4N) with improved pharmacokinetics with respect to the native protein. Considering that low productivity is the major problem of the production of recombinant proteins in mammalian cells, we designed a new strategy to generate stable cell lines with high IFN 4N expression. Lentiviral vectors containing the cDNA of IFN-4N in their genome were assembled into HEK-293 cells and employed for transduction of CHO.K1 cells. Cells were then subjected to a selection process using increasing concentrations of puromycin. Successive increases in specific productivity of IFN-4N were observed after each selection step. Thus, the cell line resistant to the highest concentration of the antibiotic exhibited a 10-fold higher specific productivity compared to the one obtained after the first selection event. Furthermore, its productivity was 10 times higher than that obtained using conventional plasmidic vectors, representing an attractive alternative for high yield production of the cytokine.