Affinity purification of streptavidin using tobacco mosaic virus particles as purification tags

Negrouk, Valentine; Eisner, Galina; Midha, Sunita; Lee, Hyung‐il; Bascomb, Newell F.; Gleba, Yuri

doi:10.1016/j.ab.2004.04.021

Cited by 8 publications

(5 citation statements)

References 14 publications

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“…Despite this knowledge, attempts by numerous groups to create new products based on protein fusions have so far met with limited success. CP fusion proteins carrying an up to 20-aa extension to the C-terminus or a 12-aa insertion in a surface loop of the CP were shown to assemble into infectious virions, with the displayed peptides ranging from neuropeptides (5,8), to vaccine epitopes (9-11), to affinity tags (12). However, addition of larger peptides at the C-terminus of TMV CP always proved to be assembly-defective.…”

mentioning

confidence: 99%

Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A

Werner¹,

Marillonnet²,

Hause³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Earlier attempts to express peptides longer than 20 aa on the surface of tobamoviruses such as tobacco mosaic virus have failed. Surprisingly, we found that a functional fragment of protein A (133 aa) can be displayed on the surface of a tobamovirus as a Cterminal fusion to the coat protein via a 15-aa linker. The macromolecular nature of these nanoparticles allowed the design of a simple protocol for purification of mAbs with a recovery yield of 50% and >90% product purity. The extremely dense packing of protein A on the nanoparticles (>2,100 copies per viral particle) results in an immunoadsorbent with a binding capacity of 2 g mAb per g. This characteristic, combined with the high level of expression of the nanoparticles (>3 g adsorbent per kg of leaf biomass), provides a very inexpensive self-assembling matrix that could meet the criteria for a single-use industrial immunoadsorbent for antibody purification.antibody purification ͉ coat protein fusion ͉ viral particle

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mentioning

confidence: 99%

Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A

Werner¹,

Marillonnet²,

Hause³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

122

123

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“…For example, recombinant TMV carrying streptavidin-specific peptide (TLIAHPQ) on the surface was used for the purification of streptavidin from Streptomyces avidinii [57]. The particles containing a fragment of protein A from Staphylococcus aureus were successfully used for the isolation of antibodies from plant extracts [81].…”

Section: Methods Of Genetic Modification Of Tmv Particlesmentioning

confidence: 99%

Chimeric particles of tobacco mosaic virus as a platform for the development of next-generation nanovaccines

2016

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The production of vaccines and other proteins in plants for medical purposes offers a number of advantages over other expression systems. The tobacco mosaic virus (TMV) is an appropriate model for the development of a variety of vectors, including those used for the assembly of chimeric particles carrying heterologous peptides on the surface and inducing an effective immune response. To overcome the problems arising during the assembly of such particles from recombinant subunits of the coat protein, peptide linkers, suppressed stop codons, and proteolytic sites are used. To date, it has been shown that TMV-based nanovaccines provide protection against the viruses of influenza A, foot-and-mouth disease, papilloma, and they are also able to overcome B-cell tolerance for cancer-cell suppression. Genetically modified TMV-based virions can accommodate the ions of various metals and act as affinity agents for protein purification.

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“…In other cases, though, the choice of a specific protocol might also reflect the ready-to-hand literature, the availability of a certain PEG type in the cupboard, or personal experience -in order to not change a winning team (Materials and Methods sections typically do not contain too many arguments). Alternative strategies for the preparation of tobamovirus particles are not extinct either: Some researchers precipitate these by ammonium sulfate [196][197][198], others refrain from any precipitation and enrich virions by different consecutive centrifugation steps. If a background of accompanying compounds from plants is tolerated or even wanted in the work, such protocols can be quite simple [199].…”

Section: |3|3 Novel Applications: Increasing Demand: Tobamovirus Part...mentioning

confidence: 99%

Getting Hold of the Tobamovirus Particle – Why and How? Purification Routes over Time and a New Customizable Approach

Wendlandt,

Britz,

Kleinow

et al. 2024

Preprint

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This article develops a multi-perspective view on motivations and methods for tobamovirus purification through the ages, and presents a novel efficient, easy-to-use approach that can be well-adapted to different species of native and functionalized virions. We survey the various driving forces prompting researchers to enrich tobamoviruses: from the search for the causative agents of mosaic diseases in plants to their increasing recognition as versatile nanocarriers in biomedical and engineering applications. Best practice and also rarely applied options for the serial processing steps required for successful isolation of tobamoviruses are then reviewed. Adaptations for distinct particle species, pitfalls and 'forgotten' or underrepresented technologies are considered as well. The article is topped off with our own development of a method for virion preparation, rooted in historical protocols. It combines selective re-solubilization of polyethylene glycol (PEG) virion raw precipitates with density step gradient centrifugation in biocompatible iodixanol formulations, yielding ready-to-use particle suspensions. This newly established protocol and some considerations for perhaps worthwile further developments could serve as putative stepping stones towards preparation procedures appropriate for routine practical uses of the multivalent soft-matter nanorods.

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Affinity purification of streptavidin using tobacco mosaic virus particles as purification tags

Cited by 8 publications

References 14 publications

Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A

Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A

Chimeric particles of tobacco mosaic virus as a platform for the development of next-generation nanovaccines

Getting Hold of the Tobamovirus Particle – Why and How? Purification Routes over Time and a New Customizable Approach

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