Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Georg Tscheuschner
ORCID logo ,
Marco Ponader
,
Christopher Raab
,
Prisca S. Weider
,
Reni Hartfiel
,
Jan Ole Kaufmann
,
Jule L. Völzke
,
Gaby Bosc-Bierne
,
Carsten Prinz
,
Timm Schwaar
,
Paul Andrle
,
Henriette Bäßler
,
Khoa Nguyen
,
Yanchen Zhu
,
Antonia S. J. S. Mey
,
Amr Mostafa
,
Ilko Bald
ORCID logo ,
Michael G. Weller
* ORCID logo
Version 1 : Received: 26 January 2023 / Approved: 30 January 2023 / Online: 30 January 2023 (10:09:14 CET)

A peer-reviewed article of this Preprint also exists.

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. Efficient Purification of Cowpea Chlorotic Mottle Virus by a Novel Peptide Aptamer. Viruses 2023, 15, 697. 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. Efficient Purification of Cowpea Chlorotic Mottle Virus by a Novel Peptide Aptamer. Viruses 2023, 15, 697.

Abstract

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 is a key step. 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 CCMV from infected plant tissue was developed, focusing on efficiency, economy, and final purity. The protocol involves precipitation with PEG 8000, followed by an 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. 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.

Keywords

Cowpea chlorotic mottle virus; purification; affinity extraction; affinity chromatography, CCMV-binding peptide; virus-like particles; VLP, plant virus, nanotechnology, nanoparticles; virus production; safety issues; ultracentrifugation-free protocol; molecular dynamics

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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