Development of pepper vein banding virus chimeric virus-like particles for potential diagnostic and therapeutic applications

Sabharwal, Pallavi; Sushmitha, C.; Amritha, Chemmenchery K; Natraj, Usha; Murthy, M.R.N.; Savithri, H.S.

doi:10.1007/s00705-020-04581-y

Cited by 6 publications

(3 citation statements)

References 47 publications

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“…The CP fusion yielded IgG-capturing flexuous VLPs [ 143 ]. Similarly, PA domain B-fused pepper vein banding virus (PVBV) CP assembled into Potyvirus -based filaments with high IgG binding capacity, intended for biomedical antibody delivery [ 144 ]. Whereas these CPs of elongated plant viruses were endowed with the PA domains at a surface-exposed terminus, another CP—of the spherical Sesbania mosaic virus (SeMV, genus Sobemovirus )—was engineered into a replacement mutant, with the PA domain B in place of a disordered CP region [ 145 ].…”

Section: Discussionmentioning

confidence: 99%

Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System

Wendlandt,

Koch,

Britz

et al. 2023

Viruses

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Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.

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Section: Discussionmentioning

confidence: 99%

Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System

Wendlandt,

Koch,

Britz

et al. 2023

Viruses

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“…Also, PVBV CP could self assemble to form VLPs in sucrose density gradient [41,47]. It was shown that the presence of extra amino acids in the N terminal (either due to cloning strategy or chimeric VLP) did not hamper the assembly [48]. However, BBrMV fusion protein with GST tag did not assemble into VLPs (data not shown) possibly because the tag is a huge peptide of 25 kDa.…”

Section: Assessment Of Bbrmv Cp Self-assembly Through Ultracentrifugamentioning

confidence: 96%

Recombinant Coat Protein of Banana Bract Mosaic Virus as a Potential Antigen for Serological Detection of the Virus

Dilip

Louis

Sabharwal

et al. 2020

CJAST

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Banana bract mosaic disease caused by Banana bract mosaic potyvirus (BBrMV) is reported to instigate heavy loss in banana and plantain across Asia. Almost all the cultivars of banana succumb to the disease resulting in malformed bunches weighing less than half of normal ones. In the current study the coat protein (CP) gene segment present at the 3’ terminal region of the viral genome amplified by RT-PCR was cloned into expression vectors, pRSET-C and pGEX-4T-2 to use it for raising polyclonal antiserum which in turn will aid in developing assays to detect the virus. Recombinant BBrMV CP (rCP) in pRSET-C when expressed was insoluble whereas, it was in the soluble fraction when expressed from pGEX-4T-2. The GST-fusion protein was purified by GSH sepharose affinity column chromatography and western blot analysis was performed using anti GST antibodies. 360 µg/ml of protein was purified from 1 l of culture. The GST tag was cleaved from the purified protein by incubation with thrombin at 25°C overnight. The rCP was characterized using ultracentrifugation, fluorescence spectroscopy and electron microscopy. The tagless monomer failed to assemble to virus like particles (VLPs) in vitro which was substantiated by fluorescence spectroscopy. This study will be first step towards deciphering structure and functions of Banana bract mosaic virus coat protein.

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“…Thus, antibodies to intracellular antigens specific to disease as therapeutics are largely unexplored. With a view to develop PVBV VLP-based nanocarriers for the delivery of antibodies, the B domain of Staphylococcus aureus protein A (SpA) (that binds to the constant region of the antibodies) was genetically engineered at the N-terminus of PVBV CP [103]. PVBV CP N-terminal fusion was designated as BCP and assembled virus particles as chimeric PVBV VLPs.…”

Section: Applications Of Potyviruses In Biotechnology: Characterizatimentioning

confidence: 99%

Functional Characterization of Pepper Vein Banding Virus-Encoded Proteins and Their Interactions: Implications in Potyvirus Infection

Sabharwal

Savithri

2020

Viruses

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Pepper vein banding virus (PVBV) is a distinct species in the Potyvirus genus which infects economically important plants in several parts of India. Like other potyviruses, PVBV encodes multifunctional proteins, with several interaction partners, having implications at different stages of the potyviral infection. In this review, we summarize the functional characterization of different PVBV-encoded proteins with an emphasis on their interaction partners governing the multifunctionality of potyviral proteins. Intrinsically disordered domains/regions of these proteins play an important role in their interactions with other proteins. Deciphering the function of PVBV-encoded proteins and their interactions with cognitive partners will help in understanding the putative mechanisms by which the potyviral proteins are regulated at different stages of the viral life-cycle. This review also discusses PVBV virus-like particles (VLPs) and their potential applications in nanotechnology. Further, virus-like nanoparticle-cell interactions and intracellular fate of PVBV VLPs are also discussed.

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Development of pepper vein banding virus chimeric virus-like particles for potential diagnostic and therapeutic applications

Cited by 6 publications

References 47 publications

Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System

Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System

Recombinant Coat Protein of Banana Bract Mosaic Virus as a Potential Antigen for Serological Detection of the Virus

Functional Characterization of Pepper Vein Banding Virus-Encoded Proteins and Their Interactions: Implications in Potyvirus Infection

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