scholarly journals Generation of Recombinant Porcine Parvovirus Virus-Like Particles inSaccharomyces cerevisiaeand Development of Virus-Specific Monoclonal Antibodies

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Paulius Lukas Tamošiūnas ◽  
Rasa Petraitytė-Burneikienė ◽  
Rita Lasickienė ◽  
Artiomas Akatov ◽  
Gabrielis Kundrotas ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Self Assembly ◽  
Gradient Centrifugation ◽  
Expression System ◽  
Porcine Parvovirus ◽  
Virus Like Particles ◽  
Vp2 Protein ◽  
Nucleic Acid Extract ◽  
Gene Coding ◽  
Electron Microscopy Analysis

Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the major capsid protein VP2 of PPV was amplified using viral nucleic acid extract from swine serum and inserted into yeastSaccharomyces cerevisiaeexpression plasmid. Recombinant PPV VP2 protein was efficiently expressed in yeast and purified using density gradient centrifugation. Electron microscopy analysis of purified PPV VP2 protein revealed the self-assembly of virus-like particles (VLPs). Nine monoclonal antibodies (MAbs) against the recombinant PPV VP2 protein were generated. The specificity of the newly generated MAbs was proven by immunofluorescence analysis of PPV-infected cells. Indirect IgG ELISA based on the recombinant VLPs for detection of PPV-specific antibodies in swine sera was developed and evaluated. The sensitivity and specificity of the new assay were found to be 93.4% and 97.4%, respectively. In conclusion, yeastS. cerevisiaerepresents a promising expression system for generating recombinant PPV VP2 protein VLPs of diagnostic relevance.

scholarly journals Virus-Like Particles Produced Using the Brome Mosaic Virus Recombinant Capsid Protein Expressed in a Bacterial System

2021 ◽  
Vol 22 (6) ◽  
pp. 3098
Author(s):  
Aleksander Strugała ◽  
Jakub Jagielski ◽  
Karol Kamel ◽  
Grzegorz Nowaczyk ◽  
Marcin Radom ◽  
...  
Keyword(s):  
Ionic Strength ◽  
Mosaic Virus ◽  
Capsid Protein ◽  
Self Assembly ◽  
Environmental Changes ◽  
Expression System ◽  
Brome Mosaic Virus ◽  
Virus Like Particles ◽  
Assembly Principles ◽  
The Impact

Virus-like particles (VLPs), due to their nanoscale dimensions, presence of interior cavities, self-organization abilities and responsiveness to environmental changes, are of interest in the field of nanotechnology. Nevertheless, comprehensive knowledge of VLP self-assembly principles is incomplete. VLP formation is governed by two types of interactions: protein–cargo and protein–protein. These interactions can be modulated by the physicochemical properties of the surroundings. Here, we used brome mosaic virus (BMV) capsid protein produced in an E. coli expression system to study the impact of ionic strength, pH and encapsulated cargo on the assembly of VLPs and their features. We showed that empty VLP assembly strongly depends on pH whereas ionic strength of the buffer plays secondary but significant role. Comparison of VLPs containing tRNA and polystyrene sulfonic acid (PSS) revealed that the structured tRNA profoundly increases VLPs stability. We also designed and produced mutated BMV capsid proteins that formed VLPs showing altered diameters and stability compared to VLPs composed of unmodified proteins. We also observed that VLPs containing unstructured polyelectrolyte (PSS) adopt compact but not necessarily more stable structures. Thus, our methodology of VLP production allows for obtaining different VLP variants and their adjustment to the incorporated cargo.

The immunogenicity of the virus-like particles derived from the VP2 protein of porcine parvovirus

2020 ◽  
Vol 248 ◽  
pp. 108795
Author(s):  
Tao Hua ◽  
Daohua Zhang ◽  
Bo Tang ◽  
Chen Chang ◽  
Guoyang Liu ◽  
...  
Keyword(s):  
Porcine Parvovirus ◽  
Virus Like Particles ◽  
Vp2 Protein

scholarly journals Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies

2014 ◽  
Vol 185 ◽  
pp. S39
Author(s):  
Rasa Petraityte Burneikiene ◽  
Paulius Lukas Tamosiunas ◽  
Artiomas Akatov ◽  
Rita Lasickiene ◽  
Gabrielis Kundrotas ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Monoclonal Antibodies ◽  
Porcine Parvovirus ◽  
Virus Like Particles

Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles.

1993 ◽  
Vol 67 (12) ◽  
pp. 6929-6936 ◽  
Author(s):  
R Kirnbauer ◽  
J Taub ◽  
H Greenstone ◽  
R Roden ◽  
M Dürst ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Self Assembly ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16

scholarly journals Self-Assembly and Release of Peste des Petits Ruminants Virus-Like Particles in an Insect Cell-Baculovirus System and Their Immunogenicity in Mice and Goats

PLoS ONE ◽  
2014 ◽  
Vol 9 (8) ◽  
pp. e104791 ◽  
Author(s):  
Wenchao Li ◽  
Hongyan Jin ◽  
Xiukun Sui ◽  
Zhanzhong Zhao ◽  
Chenghuai Yang ◽  
...  
Keyword(s):  
Self Assembly ◽  
Insect Cell ◽  
Peste Des Petits Ruminants ◽  
Virus Like Particles ◽  
Baculovirus System

scholarly journals Rotavirus Virus-Like Particles as Surrogates in Environmental Persistence and Inactivation Studies

2004 ◽  
Vol 70 (7) ◽  
pp. 3904-3909 ◽  
Author(s):  
Santiago Caballero ◽  
F. Xavier Abad ◽  
Fabienne Loisy ◽  
Françoise S. Le Guyader ◽  
Jean Cohen ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Uv Light ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Virus Removal ◽  
Virus Like Particles ◽  
Reverse Transcription Pcr ◽  
Actual Field ◽  
Uv Dose ◽  
Green Fluorescent

ABSTRACT Virus-like particles (VLPs) with the full-length VP2 and VP6 rotavirus capsid proteins, produced in the baculovirus expression system, have been evaluated as surrogates of human rotavirus in different environmental scenarios. Green fluorescent protein-labeled VLPs (GFP-VLPs) and particles enclosing a heterologous RNA (pseudoviruses), whose stability may be monitored by flow cytometry and antigen capture reverse transcription-PCR, respectively, were used. After 1 month in seawater at 20°C, no significant differences were observed between the behaviors of GFP-VLPs and of infectious rotavirus, whereas pseudovirus particles showed a higher decay rate. In the presence of 1 mg of free chlorine (FC)/liter both tracers persisted longer in freshwater at 20°C than infectious viruses, whereas in the presence of 0.2 mg of FC/liter no differences were observed between tracers and infectious rotavirus at short contact times. However, from 30 min of contact with FC onward, the decay of infectious rotavirus was higher than that of recombinant particles. The predicted Ct value for a 90% reduction of GFP-VLPs or pseudoviruses induces a 99.99% inactivation of infectious rotavirus. Both tracers were more resistant to UV light irradiation than infectious rotavirus in fresh and marine water. The effect of UV exposure was more pronounced on pseudovirus than in GFP-VLPs. In all types of water, the UV dose to induce a 90% reduction of pseudovirus ensures a 99.99% inactivation of infectious rotavirus. Recombinant virus surrogates open new possibilities for the systematic validation of virus removal practices in actual field situations where pathogenic agents cannot be introduced.

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