Downstream processing of triple layered rotavirus like particles.

Area of Science:

• Biotechnology
• Vaccine Development
• Virology

Background:

• Rotavirus disease causes significant child mortality globally.
• Current purification methods for rotavirus like particles (RLPs) are not scalable or cost-effective.
• Pre-clinical trial material demands improved RLP purification strategies.

Purpose of the Study:

• To develop a reproducible, scalable, and cost-effective purification process for rotavirus like particles (RLPs).
• To assess the stability of purified RLPs in different formulations.
• To provide an alternative to traditional ultracentrifugation methods for VLP purification.

Main Methods:

• Production of virus like particles (VLPs) composed of VP2, VP6, and VP7 rotavirus proteins using an insect cell/baculovirus system at a 5L scale.
• Stepwise purification utilizing depth filtration, ultrafiltration, and size exclusion chromatography.
• Assessment of particle stability using size exclusion chromatography and formulation studies with glycerol or trehalose.

Main Results:

• A purification process achieved 37% overall recovery yield.
• Demonstrated removal of 95% of host cell proteins and 99% of host cell DNA.
• Formulations with glycerol or trehalose maintained 75% intact triple-layered VLPs at 4°C for 4 months.

Conclusions:

• The developed purification strategy is a promising alternative for downstream processing of VLPs.
• This method offers a scalable and cost-effective approach for producing RLPs for vaccine development.
• Optimized formulations enhance the stability of purified rotavirus particles.