Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies
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 yeast Saccharomyces cerevisiae...
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| Format: | Article |
| Language: | English |
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Wiley
2014-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2014/573531 |
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| author | Paulius Lukas Tamošiūnas Rasa Petraitytė-Burneikienė Rita Lasickienė Artiomas Akatov Gabrielis Kundrotas Vilimas Sereika Raimundas Lelešius Aurelija Žvirblienė Kęstutis Sasnauskas |
| author_facet | Paulius Lukas Tamošiūnas Rasa Petraitytė-Burneikienė Rita Lasickienė Artiomas Akatov Gabrielis Kundrotas Vilimas Sereika Raimundas Lelešius Aurelija Žvirblienė Kęstutis Sasnauskas |
| author_sort | Paulius Lukas Tamošiūnas |
| collection | DOAJ |
| description | 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 yeast Saccharomyces cerevisiae expression 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, yeast S. cerevisiae represents a promising expression system for generating recombinant PPV VP2 protein VLPs of diagnostic relevance. |
| format | Article |
| id | doaj-art-47f3a9dcb4c5453cb1a04a3d2e75fb55 |
| institution | Kabale University |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-47f3a9dcb4c5453cb1a04a3d2e75fb552025-02-03T05:43:33ZengWileyJournal of Immunology Research2314-88612314-71562014-01-01201410.1155/2014/573531573531Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal AntibodiesPaulius Lukas Tamošiūnas0Rasa Petraitytė-Burneikienė1Rita Lasickienė2Artiomas Akatov3Gabrielis Kundrotas4Vilimas Sereika5Raimundas Lelešius6Aurelija Žvirblienė7Kęstutis Sasnauskas8Institute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaInstitute of Microbiology and Virology, Veterinary Faculty of Veterinary Academy, Lithuanian University of Health Sciences, Tilžės 18, 47181 Kaunas, LithuaniaInstitute of Microbiology and Virology, Veterinary Faculty of Veterinary Academy, Lithuanian University of Health Sciences, Tilžės 18, 47181 Kaunas, LithuaniaInstitute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241 Vilnius, LithuaniaPorcine 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 yeast Saccharomyces cerevisiae expression 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, yeast S. cerevisiae represents a promising expression system for generating recombinant PPV VP2 protein VLPs of diagnostic relevance.http://dx.doi.org/10.1155/2014/573531 |
| spellingShingle | Paulius Lukas Tamošiūnas Rasa Petraitytė-Burneikienė Rita Lasickienė Artiomas Akatov Gabrielis Kundrotas Vilimas Sereika Raimundas Lelešius Aurelija Žvirblienė Kęstutis Sasnauskas Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies Journal of Immunology Research |
| title | Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies |
| title_full | Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies |
| title_fullStr | Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies |
| title_full_unstemmed | Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies |
| title_short | Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies |
| title_sort | generation of recombinant porcine parvovirus virus like particles in saccharomyces cerevisiae and development of virus specific monoclonal antibodies |
| url | http://dx.doi.org/10.1155/2014/573531 |
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