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Cited by 10Transgenic Plants and ApplicationsPlant Virus Research StudiesProtein purification and stability

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Efficient Purification of Cowpea Chlorotic Mottle Virus by a Novel Peptide Aptamer

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Tscheuschner, Georg, Ponader, Marco, Raab, Christopher, Weider, Prisca S., Hartfiel, Reni, Kaufmann, Jan Ole, Völzke, Jule L., Bosc-Bierne, Gaby, Prinz, Carsten, Schwaar, Timm, Andrle, Paul, Bäßler, Henriette, Nguyen, Khoa, Zhu, Yanchen, Mey, Antonia S. J. S., Mostafa, Amr, Bald, Ilko, Weller, Michael G.·2023·Viruses
DOI 10.3390/v15030697PMC PMC10051510

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Abstract

The cowpea chlorotic mottle virus (CCMV) is a plant virus explored as a nanotechnological platform. The robust self-assembly mechanism of its capsid protein allows for drug encapsulation and targeted delivery. Additionally, the capsid nanoparticle can be used as a programmable platform to display different molecular moieties. In view of future applications, efficient production and purification of plant viruses are key steps. In established protocols, the need for ultracentrifugation is a significant limitation due to cost, difficult scalability, and safety issues. In addition, the purity of the final virus isolate often remains unclear. Here, an advanced protocol for the purification of the CCMV from infected plant tissue was developed, focusing on efficiency, economy, and final purity. The protocol involves precipitation with PEG 8000, followed by affinity extraction using a novel peptide aptamer. The efficiency of the protocol was validated using size exclusion chromatography, MALDI-TOF mass spectrometry, reversed-phase HPLC, and sandwich immunoassay. Furthermore, it was demonstrated that the final eluate of the affinity column is of exceptional purity (98.4%) determined by HPLC and detection at 220 nm. The scale-up of our proposed method seems to be straightforward, which opens the way to the large-scale production of such nanomaterials. This highly improved protocol may facilitate the use and implementation of plant viruses as nanotechnological platforms for in vitro and in vivo applications.

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Metadata

Authors
Tscheuschner, G., Ponader, M., Raab, C., Weider, P. S., Hartfiel, R., Kaufmann, J. O., Völzke, J. L., Bosc-Bierne, G., Prinz, C., Schwaar, T., Andrle, P., Bäßler, H., Nguyen, K., Zhu, Y., Mey, A. S. J. S., Mostafa, A., Bald, I., & Weller, M. G.
Affiliations
  • Federal Institute For Materials Research and Testing (DE)
  • Rockwool Foundation Berlin (DE)
  • University of Edinburgh (GB)
  • University of Potsdam (DE)
Corresponding author
Weller, Michael G.
ORCID
Venue
Viruses
Publisher
MDPI AG
Published
2023-03-07
Volume
15
Issue
3
Pages
697
ISSN
1999-4915 (ISSN-L)
DOI
10.3390/v15030697
PMC
PMC10051510
OpenAlex
W4323568210
Cited by
10
References
2
Citations by year
2026: 5 · 2025: 2 · 2024: 1 · 2023: 2
FWCI
1.29
Type
journal article
Language
en
Open access
Licence
Locations
  • Viruses (publishedVersion, cc-by)
  • Preprints.org (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Edinburgh Research Explorer (University of Edinburgh) (submittedVersion, cc-by)
Funders
Université Paris-Saclay
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Virus Research Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
Keywords
cowpea chlorotic mottle virus, peptide aptamers, drug encapsulation, targeted delivery, scale-up production
MeSH terms
Capsid, Bromovirus, Capsid Proteins, Aptamers, Peptide, Nanoparticles

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