Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells

TARGOVNIK, ALEXANDRA MARISA; FERRARI, ALEJANDRO; MC CALLUM, GREGORIO JUAN; ARREGUI, MARIANA BERNADETT; SMITH, IGNACIO; BRACCO, LAUTARO FIDEL; ALFONSO, VICTORIA; LÓPEZ, MARÍA GABRIELA; MARTÍNEZ-SOLÍS, MARÍA; HERRERO, SALVADOR; MIRANDA, MARÍA VICTORIA

3 Biotech

SPRINGER

Año: 2019 vol. 9

2190-572X

In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabiesvirus glycoprotein G (GE) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vectorsystem in Sf9 insect cells combined with a novel chimeric promoter (polh-pSeL), the expression level reached a yield of4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard polh promoter alone. The proteinwas recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yieldof 95% and a purity of 87%. Finally, GEwas successfully used in an assay to detect specific antibodies in serum samplesderived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to producehigh amounts of this glycoprotein.