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

Content Provider	Europe PMC
Author	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.
Editor	Yan, Fei
Copyright Year	2023
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.
Journal	Viruses
Volume Number	15
PubMed Central reference number	PMC10051510
Issue Number	3
PubMed reference number	36992405
e-ISSN	19994915
DOI	10.3390/v15030697
Language	English
Publisher	MDPI
Publisher Date	2023-03-07
Access Restriction	Open
Rights License	Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). © 2023 by the authors.
Subject Keyword	cowpea chlorotic mottle virus purification affinity extraction affinity chromatography CCMV-binding peptide virus-like particles plant virus nanotechnology nanoparticles virus production safety issues ultracentrifugation-free protocol molecular dynamics
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Virology