info:eu-repo/semantics/article
Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells
Fecha
2019-10Registro en:
Targovnik, Alexandra Marisa; Ferrari, Alejandro; Mc Callum, Gregorio Juan; Arregui, Mariana Bernadett; Smith, Ignacio; et al.; Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells; Springer; 3 Biotech; 9; 11; 10-2019; 1-11
2190-572X
2190-5738
CONICET Digital
CONICET
Autor
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
Resumen
In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (GE) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (polh-pSeL), the expression level reached a yield of 4.1± 0.3 mg/L culture, which was signifcantly higher than that achieved with the standard polh promoter alone. The protein was recovered from the cell lysates and easily purifed in only one step by metal ion afnity chromatography, with a yield of 95% and a purity of 87%. Finally, GE was successfully used in an assay to detect specifc antibodies in serum samplesderived from rabies-vaccinated animals. The efcient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein.