Production and characterization of chimeric SARS-CoV-2 antigens based on the capsid protein of cowpea chlorotic mottle virus

Almendárez-Rodriguez, Claudia; Solís-Andrade, Karla I.; Govea‐Alonso, Dania O.; Comas-García, Mauricio; Rosales‐Mendoza, Sergio

doi:10.1016/j.ijbiomac.2022.06.021

Cited by 6 publications

(4 citation statements)

References 44 publications

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“…Apart from heterologous RNAs, a wide range of other non-viral cargo has also been packaged by self-assembled viral capsid proteins of CCMV; for example, anionic polymers [ 25 ], mineralized salts [ 26 ], gold nanoparticles [ 27 ], and fluorescent proteins [ 28 ]. In addition, CCMV VLPs were recently studied as potential candidates for in situ cancer vaccines [ 29 ] and scaffolds to display SARS-CoV-2 antigens [ 30 ].…”

Section: Introductionmentioning

confidence: 99%

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

Tscheuschner

Ponader

Raab

et al. 2023

Viruses

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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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Section: Introductionmentioning

confidence: 99%

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

Tscheuschner

Ponader

Raab

et al. 2023

Viruses

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show abstract

“…It can take several weeks before the antigen is available for immunization. Many antigens are produced in intracellular compartments, which require isolation by cellular lysis, purification by chromatography, and often solubilization or refolding in order to obtain the desired protein conformation [ 6–9 ]. Reduction of process steps for antigen production will reduce overall the costs of manufacturing mAbs [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

tANCHOR fast and cost-effective cell-based immunization approach with focus on the receptor-binding domain of SARS-CoV-2

Bernauer,

Schlör,

Maier

et al. 2023

Biology Methods and Protocols

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Successful induction of antibodies in model organisms like mice depends strongly on antigen design and delivery. New antigen designs for immunization are helpful for developing future therapeutic monoclonal antibodies (mAbs). One of the gold standards to induce antibodies in mice is to express and purify the antigen for vaccination. This is especially time-consuming when mAbs are needed rapidly. We closed this gap and used the display technology tetraspanin anchor to develop a reliable immunization technique without the need to purify the antigen. This technique is able to speed up the immunization step enormously and we have demonstrated that we were able to induce antibodies against different proteins with a focus on the receptor-binding domain of SARS-CoV-2 and the extracellular loop of canine cluster of differentiation 20 displayed on the surface of human cells.

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“…Apart from heterologous RNAs, a wide range of other non-viral cargo has also been packaged by self-assembled viral capsid proteins of CCMV, for example, anionic polymers [24], mineralized salts [25], gold nanoparticles [26], and fluorescent proteins [27]. In addition, CCMV VLPs were recently studied as potential candidates for in situ cancer vaccines [28] and scaffolds to display SARS-CoV-2 antigens [29].…”

Section: Introductionmentioning

confidence: 99%

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

Tscheuschner¹,

Ponader²,

Raab³

et al. 2023

Preprint

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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.

show abstract

Production and characterization of chimeric SARS-CoV-2 antigens based on the capsid protein of cowpea chlorotic mottle virus

Cited by 6 publications

References 44 publications

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

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

tANCHOR fast and cost-effective cell-based immunization approach with focus on the receptor-binding domain of SARS-CoV-2

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

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