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.
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Acknowledgements
This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica de Argentina (PICT 2015-1992; PICT 2014-3350), by Universidad de Buenos Aires (UBACyT 2016-20020150100145BA). AMT, VA, MGL, AF, and MVM are career researchers of the Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET). GJM, MBA, and IS are research fellows of CONICET.
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AMT, SH, AF, and MVM designed the experiments. AMT and MMS constructed the baculovirus vectors. AMT and GJM performed the expression experiments. AMT and LFB contributed to downstream processing. IS and MBA performed the characterization experiments. AF contributed to the development of the ELISA. VA and MGL performed the confocal analysis. AMT, AF, SH, and MVM analyzed the data. All authors discussed the results and agreed upon the final manuscript.
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Targovnik, A.M., Ferrari, A., Mc Callum, G.J. et al. Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells. 3 Biotech 9, 385 (2019). https://doi.org/10.1007/s13205-019-1920-4
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DOI: https://doi.org/10.1007/s13205-019-1920-4