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An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris

An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris

Authors :
Sebastian Fernando Cavalitto
Marco Antonio Tizzano
Guillermo Hernán Sguazza
Leandro Daniel Picotto
Marcelo Ricardo Ítalo Pecoraro
Cecilia Mónica Galosi
Source :
SEDICI (UNLP), Universidad Nacional de La Plata, instacron:UNLP
Publication Year :
2017
Publisher :
Academic Press Inc Elsevier Science, 2017.

Abstract

The glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted protein, using a simplified DO-stat control feeding strategy. This strategy involves the addition of methanol when the dissolved oxygen (DO) level rises above the setpoint avoiding methanol accumulation and oxygen limitation. The G-protein expression was evaluated by SDS-PAGE, ELISA, and western blot assays. Like native G-protein, the recombinant G-protein was found reactive when it was challenged against specific antibodies. The data indicate that the recombinant G-protein can be easily expressed and isolated, and may be useful as a safe source in the production of diagnostic kits and subunit vaccines to prevent rabies.<br />Facultad de Ciencias Veterinarias<br />Centro de Investigación y Desarrollo en Fermentaciones Industriales

Details

Language :
English
Database :
OpenAIRE
Journal :
SEDICI (UNLP), Universidad Nacional de La Plata, instacron:UNLP
Accession number :
edsair.doi.dedup.....c455ddf86960c2d1bc562df5b0560258