Optimizing the synthesis and purification of MS2 virus like particles
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 s...
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doaj-0d1c8ffd1d7f4708bef03beb07f183f22021-10-10T11:30:38ZengNature Publishing GroupScientific Reports2045-23222021-10-0111111410.1038/s41598-021-98706-1Optimizing the synthesis and purification of MS2 virus like particlesKhadijeh Hashemi0Mohammad Mahdi Ghahramani Seno1Mohammad Reza Ahmadian2Bizhan Malaekeh-Nikouei3Mohammad Reza Bassami4Hesam Dehghani5Amir Afkhami-Goli6Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of MashhadDivision of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of MashhadInstitute of Biochemistry and Molecular Biology II, Medical Faculty of the Heinrich-Heine UniversityNanotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical SciencesDivision of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of MashhadDivision of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of MashhadDivision of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of MashhadAbstract 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.https://doi.org/10.1038/s41598-021-98706-1 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Khadijeh Hashemi Mohammad Mahdi Ghahramani Seno Mohammad Reza Ahmadian Bizhan Malaekeh-Nikouei Mohammad Reza Bassami Hesam Dehghani Amir Afkhami-Goli |
spellingShingle |
Khadijeh Hashemi Mohammad Mahdi Ghahramani Seno Mohammad Reza Ahmadian Bizhan Malaekeh-Nikouei Mohammad Reza Bassami Hesam Dehghani Amir Afkhami-Goli Optimizing the synthesis and purification of MS2 virus like particles Scientific Reports |
author_facet |
Khadijeh Hashemi Mohammad Mahdi Ghahramani Seno Mohammad Reza Ahmadian Bizhan Malaekeh-Nikouei Mohammad Reza Bassami Hesam Dehghani Amir Afkhami-Goli |
author_sort |
Khadijeh Hashemi |
title |
Optimizing the synthesis and purification of MS2 virus like particles |
title_short |
Optimizing the synthesis and purification of MS2 virus like particles |
title_full |
Optimizing the synthesis and purification of MS2 virus like particles |
title_fullStr |
Optimizing the synthesis and purification of MS2 virus like particles |
title_full_unstemmed |
Optimizing the synthesis and purification of MS2 virus like particles |
title_sort |
optimizing the synthesis and purification of ms2 virus like particles |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-10-01 |
description |
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. |
url |
https://doi.org/10.1038/s41598-021-98706-1 |
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