Optimizing the synthesis and purification of MS2 virus like particles
| Autor: | Mohammad Reza Bassami, Bizhan Malaekeh-Nikouei, Khadijeh Hashemi, Mohammad Reza Ahmadian, Mohammad Mahdi Ghahramani Seno, Hesam Dehghani, Amir Afkhami-Goli |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Tris
Science viruses Dispersity Buffers Chemical Fractionation complex mixtures Article Cell Line Small hairpin RNA chemistry.chemical_compound In vivo Bacteriophage MS2 Humans Particle Size Levivirus HEPES Nitrates Multidisciplinary Chromatography biology Drug discovery Biological techniques Virion RNA virus diseases Hydrogen-Ion Concentration biology.organism_classification Chemical biology chemistry Drug delivery Medicine Biotechnology |
| Zdroj: | Scientific Reports Scientific Reports, Vol 11, Iss 1, Pp 1-14 (2021) |
| ISSN: | 2045-2322 |
| Popis: | Introducing bacteriophage MS2 virus-like particles (VLPs) as gene and drug delivery tools increases the demand for optimizing their production and purification procedure. PEG precipitation method is used efficiently to purify VLPs, while the effects of pH and different electrolytes on the stability, size, and homogeneity of purified MS2 VLPs, and the encapsulated RNA sequences remained to be elucidated. In this regard, a vector, capable of producing VLP with an shRNA packed inside was prepared. The resulting VLPs in different buffers/solutions were assessed for their size, polydispersity index, and ability to protect the enclosed shRNA. We report that among Tris, HEPES, and PBS, with or without NaNO3, and also NaNO3 alone in different pH and ionic concentrations, the 100 mM NaNO3-Tris buffer with pH:8 can be used as a new and optimal MS2 VLP production buffer, capable of inhibiting the VLPs aggregation. These VLPs show a size range of 27-30 nm and suitable homogeneity with minimum 12-month stability at 4 °C. Moreover, the resulting MS2 VLPs were highly efficient and stable for at least 48 h in conditions similar to in vivo. These features of MS2 VLPs produced in the newly introduced buffer make them an appropriate candidate for therapeutic agents’ delivery. |
| Databáze: | OpenAIRE |
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