EXPRESSION AND PURIFICATION OF RECOMBINANT FELINE INTERFERON IN THE BACULOVIRUS-INSECT LARVAE SYSTEM

ALEXANDRA MARISA TARGOVNIK; MARCELA SOLANGE VILLAVERDE; MARIANA ARREGUI; MARIELA A. FOGAR; OSCAR TABOGA; GERARDO CLAUDIO GLIKIN; LILIANA MARÍA ELENA FINOCCHIARO; OSVALDO CASCONE; MARÍA VICTORIA MIRANDA

PROCESS BIOCHEMISTRY

ELSEVIER

Lugar: Amsterdam; Año: 2014 vol. 49 p. 917 - 926

0032-9592

Feline interferons (FeIFNs) are cytokines with antiviral, antitumor and immunomodulatory functions used as therapeutic agents in a variety of veterinary diseases. In this work, FeIFN-α7 and FeIFN-α7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days post-infection (dpi), the concentrations of FeIFN-α7 and FeIFN-α7xArg in suspension culture were (1.28±0.15) x106 U ml-1 and (1.3±0.2) x106 U ml-1respectively. The maximum expression levels of FeIFN-α7 and FeIFN-α7xArg were (3.7±0.2) x106U ml-1 and (3.5±0.4) x106U ml-1 at 2 dpi in Rachiplusia nu larvae and (1.1±0.2) x 106 U ml-1 and (1.0±0.15) x106 U ml-1 at 6 dpi in Spodoptera frugiperda larvae respectively. R. nu was a better host for FeIFN-α7 and FeIFN-α7xArg expression. The 8xArg tag did not affect the biological activity of FeIFN- α 7 and was useful to promote the FeIFN-α7xArg adsorption on ion exchange chromatography (IEC), allowing its purification in a single step from supernatant culture and R. nu larvae. FeIFN-α7xArg was purified from the larval extract with a yield of 70% and a purification factor of 25 free of viruses. We conclude that R. nu larvae are new low-cost hosts for the expression of recombinant FeIFN-α7.