Production and characterization of active recombinant interleukin-12/eGFP fusion protein in stably-transfected DF1 XXXX cells.

@entity1 @entity1573 @entity7507 @entity3733
UNASSIGNED: The adjuvant activity of @entity1573 interleukin-12 (chIL-12) protein has been described as similar to that of @entity1 IL-12. Recombinant chIL-12 can be produced using several methods, but chIL-12 production in eukaryotic cells is lower than that in prokaryotic cells. Stimulating compounds, such as @entity3733 ( @entity3733 ), can be added to animal cell cultures to overcome this drawback. In this study, we constructed a cell line, DF1/chIL-12 which stably expressed a fusion protein, chIL-12 and enhanced green fluorescent protein (eGFP) connected by a ( @entity7507 )3 linker sequence. Fusion protein production was increased when cells were cultured in the presence of @entity3733 . When 1 * 10(6) DF1/chIL-12 cells were inoculated in a T-175 flask containing 30 mL of media, incubated for 15 h, and further cultivated in the presence of 4% @entity3733 for 48 h, the production of total fusion protein was mostly enhanced compared with the production of total fusion protein by using cell lysates induced with @entity3733 at other concentrations. The concentrations of the unpurified and purified total fusion proteins in cell lysates were 2,781    2.72 ng mL(-1) and 2,207    3.28 ng mL(-1) , respectively. The recovery rate was 79%. The fusion protein stimulated @entity1573 splenocytes to produce IFN-y, which was measured using an enzyme-linked immunosorbent assay, in the culture supernatant, indicating that treating DF1/chIL-12 cells with @entity3733 or producing chIL-12 in a fusion protein form does not have adverse effects on the bioactivity of chIL-12.    2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:641-649, 2015.
