Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system

TARGOVNIK A. ; VILLAVERDE M. S.; ARREGUI M. B.; FOGAR M. ; TABOGA O. ; GLIKIN G. C.; FINOCCHIARO L. M. E.; CASCONE O.; VICTORIA M. V.

PROCESS BIOCHEMISTRY - (Print)

ELSEVIER SCI LTD

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

1359-5113

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-a7 and FeIFN-a7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days post-infection (dpi), the concentrations of FeIFN-a7 and FeIFN-a7xArg in suspension culture were (1.28 ± 0.15) × 106 U ml-1 and (1.3 ± 0.2) × 106 U ml-1 respectively. The maximum expression levels of FeIFN-a7 and FeIFN-a7xArg were (3.7 ± 0.2) × 106 U ml-1 and (3.5 ± 0.4) × 106 U ml−1 at 2 dpi in Rachiplusia nu larvae and (1.1 ± 0.2) × 106 U ml-1 and (1.0 ± 0.15) × 106 U ml-1 at 5 dpi in Spodoptera frugiperda larvae respectively. R. nu was a better host for FeIFN-a7 and FeIFN-a7xArg expression. The 8xArg tag did not affect the biological activity of FeIFN-a7 and was useful to promote the FeIFN-a7xArg adsorption on ion exchange chromatography (IEC), allowing its purification in a single step from supernatant culture and R. nu larvae. FeIFN-a7xArg 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-a7.