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Immunization of mice against encephalomyocarditis virus. I. Purification, concentration, and inactivation of
Abstract:
Methods are described for the concentration, purification, and inactivation of encephalomyocarditis virus grown in L-cell suspension cultures. Virus concentrates were obtained by the precipitation of crude virus lysates with polyethylene glycol and were purified by Genetron extraction. After column chromatography approximately 70% recovery of infectivity and hemagglutination (HA) activity was found. Exposure of the purified virus suspensions to ultraviolet light resulted in reduction of the infectivity by a factor of 10(9) with little or no loss of HA activity. The preparations are being evaluated as vaccines.
Insights
Methods were developed to concentrate, purify, and inactivate encephalomyocarditis virus. This process achieved high recovery of infectivity and hemagglutination activity, with ultraviolet light effectively reducing infectivity for potential vaccine use.
Area of Science:
- Virology
- Biotechnology
- Vaccine Development
Background:
- Encephalomyocarditis virus (EMCV) requires effective methods for concentration and purification for research and vaccine applications.
- Existing methods may not efficiently preserve viral infectivity and activity.
Purpose of the Study:
- To establish a robust protocol for concentrating, purifying, and inactivating encephalomyocarditis virus.
- To evaluate the recovery of viral infectivity and hemagglutination (HA) activity post-purification.
- To assess the efficacy of ultraviolet (UV) light for viral inactivation while preserving HA activity.
Main Methods:
- EMCV was cultured in L-cell suspension cultures.
- Virus concentration was achieved via polyethylene glycol precipitation of crude lysates.
- Purification involved Genetron extraction and column chromatography.
- Viral inactivation was performed using ultraviolet light exposure.
Main Results:
- Polyethylene glycol precipitation and Genetron extraction yielded concentrated virus preparations.
- Column chromatography resulted in approximately 70% recovery of infectivity and hemagglutination (HA) activity.
- UV light exposure reduced viral infectivity by a factor of 10(9) with minimal loss of HA activity.
Conclusions:
- A multi-step method effectively concentrates, purifies, and inactivates EMCV.
- The developed method preserves key viral activities (infectivity and HA) while ensuring safety through inactivation.
- The purified and inactivated EMCV preparations show promise for evaluation as vaccines.