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Expression and Purification of Virus-like Particles for Vaccination
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Optimizing the synthesis and purification of MS2 virus like particles
Khadijeh Hashemi1,2, Mohammad Mahdi Ghahramani Seno1,3,4, Mohammad Reza Ahmadian5
1Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Scientific Reports
|October 7, 2021
Summary
Optimizing bacteriophage MS2 virus-like particles (VLPs) production is crucial for gene and drug delivery. A novel buffer (100 mM NaNO3-Tris, pH 8) enhances MS2 VLP stability, homogeneity, and protects encapsulated shRNA.
Area of Science:
- Biotechnology
- Nanotechnology
- Molecular Biology
Background:
- Bacteriophage MS2 virus-like particles (VLPs) are promising for gene and drug delivery.
- Optimizing VLP production and purification is essential for their application.
- The impact of buffer conditions on MS2 VLP stability and RNA protection requires further investigation.
Purpose of the Study:
- To identify optimal buffer conditions for producing stable and homogeneous MS2 VLPs.
- To evaluate the protective effect of different buffers on encapsulated shRNA.
- To determine the long-term stability and in vivo-like stability of optimized MS2 VLPs.
Main Methods:
- Preparation of a vector for producing MS2 VLPs with encapsulated shRNA.
- Purification of VLPs using PEG precipitation.
- Assessment of VLP size, polydispersity index, and shRNA protection in various buffers (Tris, HEPES, PBS) with/without NaNO3 at different pH and ionic concentrations.
Main Results:
- The 100 mM NaNO3-Tris buffer at pH 8 was identified as optimal for MS2 VLP production, preventing aggregation.
- Optimized MS2 VLPs exhibited a size range of 27-30 nm with good homogeneity.
- VLPs demonstrated a minimum 12-month stability at 4°C and were stable for at least 48 hours in simulated in vivo conditions.
Conclusions:
- The novel 100 mM NaNO3-Tris buffer (pH 8) significantly improves MS2 VLP production, stability, and homogeneity.
- These optimized MS2 VLPs effectively protect encapsulated shRNA and exhibit excellent stability.
- The findings position these MS2 VLPs as suitable candidates for therapeutic agent delivery systems.
