Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5516099 | Protein Expression and Purification | 2017 | 7 Pages |
â¢The rabies virus glycoprotein was expressed in Pichia pastoris yeast.â¢It was achieved produce viral antigen for medical use in a safe way.â¢A DO-stat control strategy reduced the culture time and cost of bioprocess.
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.