| Literature DB >> 31656723 |
Alexandra Marisa Targovnik1,2, Alejandro Ferrari3, Gregorio Juan Mc Callum1,2, Mariana Bernadett Arregui1,2, Ignacio Smith1,2, Lautaro Fidel Bracco1,2, Victoria Alfonso4,5, María Gabriela López4,5, María Martínez-Solís6, Salvador Herrero6, María Victoria Miranda1,2.
Abstract
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 significantly higher than that achieved with the standard polh promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, GE was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein. © King Abdulaziz City for Science and Technology 2019.Entities:
Keywords: Baculovirus; Rabies virus glycoprotein; Sf9 cells; polh-pSeL promoter
Year: 2019 PMID: 31656723 PMCID: PMC6778167 DOI: 10.1007/s13205-019-1920-4
Source DB: PubMed Journal: 3 Biotech ISSN: 2190-5738 Impact factor: 2.406