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

PICOTTO, LEONARDO; SGUAZZA, HERNAN; TIZZANO, MARCO; GALOSI, C.M.; CAVALITTO, SEBASTIÁN FERNANDO; PECORARO, MARCELO

PROTEIN EXPRESSION AND PURIFICATION

ACADEMIC PRESS INC ELSEVIER SCIENCE

Lugar: Amsterdam; Año: 2017 vol. 132 p. 124 - 130

1046-5928

The glycoprotein (G-protein) of rabiesvirus is responsible for viral attachment to the host cell surface and inducesvirus neutralization antibodies. In the present study, the G-protein gene ofrabies 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 strategyinvolves the addition of methanol when the dissolved oxygen (DO) level risesabove the setpoint avoiding methanol accumulation and oxygen limitation. TheG-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 waschallenged against specific antibodies. Thedata indicate that the recombinant G-protein can be easily expressed andisolated, and may be useful as a safe source in the production of diagnostickits and subunit vaccines to prevent rabies.