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Optimizing the synthesis and purification of MS2 virus like particles

Authors
  • Hashemi, Khadijeh1, 2
  • Ghahramani Seno, Mohammad Mahdi1, 1, 3
  • Ahmadian, Mohammad Reza4
  • Malaekeh-Nikouei, Bizhan5
  • Bassami, Mohammad Reza1
  • Dehghani, Hesam1, 1, 2
  • Afkhami-Goli, Amir1, 1, 2
  • 1 Ferdowsi University of Mashhad,
  • 2 Research Institute of Biotechnology, Ferdowsi University of Mashhad,
  • 3 Hospital for Sick Children,
  • 4 Medical Faculty of the Heinrich-Heine University,
  • 5 Mashhad University of Medical Sciences,
Type
Published Article
Journal
Scientific Reports
Publisher
Springer Nature
Publication Date
Oct 06, 2021
Volume
11
Identifiers
DOI: 10.1038/s41598-021-98706-1
PMCID: PMC8494748
Source
PubMed Central
Keywords
Disciplines
  • Article
License
Unknown

Abstract

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.

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