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Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies

Schmallenberg virus (SBV), discovered in continental Europe in late 2011, causes mild clinical signs in adult ruminants, including diarrhoea and reduced milk yield. However, fetal infection can lead to severe malformation in newborn offspring. To develop improved reagents for SBV serology, a high-le...

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Autores principales: Lazutka, Justas, Zvirbliene, Aurelija, Dalgediene, Indre, Petraityte-Burneikiene, Rasa, Spakova, Aliona, Sereika, Vilimas, Lelesius, Raimundas, Wernike, Kerstin, Beer, Martin, Sasnauskas, Kestutis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4058894/
https://www.ncbi.nlm.nih.gov/pubmed/24982920
http://dx.doi.org/10.1155/2014/160316
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author Lazutka, Justas
Zvirbliene, Aurelija
Dalgediene, Indre
Petraityte-Burneikiene, Rasa
Spakova, Aliona
Sereika, Vilimas
Lelesius, Raimundas
Wernike, Kerstin
Beer, Martin
Sasnauskas, Kestutis
author_facet Lazutka, Justas
Zvirbliene, Aurelija
Dalgediene, Indre
Petraityte-Burneikiene, Rasa
Spakova, Aliona
Sereika, Vilimas
Lelesius, Raimundas
Wernike, Kerstin
Beer, Martin
Sasnauskas, Kestutis
author_sort Lazutka, Justas
collection PubMed
description Schmallenberg virus (SBV), discovered in continental Europe in late 2011, causes mild clinical signs in adult ruminants, including diarrhoea and reduced milk yield. However, fetal infection can lead to severe malformation in newborn offspring. To develop improved reagents for SBV serology, a high-level yeast expression system was employed to produce recombinant SBV nucleocapsid (N) protein. Recombinant SBV N protein was investigated as an antigen in SBV-specific IgG enzyme immunoassay and used for generation of monoclonal antibodies (MAbs). Yeast-expressed SBV N protein was reactive with anti-SBV IgG-positive cow serum specimens collected from different farms of Lithuania. After immunization of mice with recombinant SBV N protein, four MAbs were generated. The MAbs raised against recombinant SBV N protein reacted with native viral nucleocapsids in SBV-infected BHK cells by immunofluorescence assay. The reactivity of recombinant N protein with SBV-positive cow serum specimens and the ability of the MAbs to recognize virus-infected cells confirm the antigenic similarity between yeast-expressed SBV N protein and native viral nucleocapsids. Our study demonstrates that yeast expression system is suitable for high-level production of recombinant SBV N protein and provides the first evidence on the presence of SBV-specific antibodies in cow serum specimens collected in Lithuania.
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spelling pubmed-40588942014-06-30 Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies Lazutka, Justas Zvirbliene, Aurelija Dalgediene, Indre Petraityte-Burneikiene, Rasa Spakova, Aliona Sereika, Vilimas Lelesius, Raimundas Wernike, Kerstin Beer, Martin Sasnauskas, Kestutis J Immunol Res Research Article Schmallenberg virus (SBV), discovered in continental Europe in late 2011, causes mild clinical signs in adult ruminants, including diarrhoea and reduced milk yield. However, fetal infection can lead to severe malformation in newborn offspring. To develop improved reagents for SBV serology, a high-level yeast expression system was employed to produce recombinant SBV nucleocapsid (N) protein. Recombinant SBV N protein was investigated as an antigen in SBV-specific IgG enzyme immunoassay and used for generation of monoclonal antibodies (MAbs). Yeast-expressed SBV N protein was reactive with anti-SBV IgG-positive cow serum specimens collected from different farms of Lithuania. After immunization of mice with recombinant SBV N protein, four MAbs were generated. The MAbs raised against recombinant SBV N protein reacted with native viral nucleocapsids in SBV-infected BHK cells by immunofluorescence assay. The reactivity of recombinant N protein with SBV-positive cow serum specimens and the ability of the MAbs to recognize virus-infected cells confirm the antigenic similarity between yeast-expressed SBV N protein and native viral nucleocapsids. Our study demonstrates that yeast expression system is suitable for high-level production of recombinant SBV N protein and provides the first evidence on the presence of SBV-specific antibodies in cow serum specimens collected in Lithuania. Hindawi Publishing Corporation 2014 2014-05-29 /pmc/articles/PMC4058894/ /pubmed/24982920 http://dx.doi.org/10.1155/2014/160316 Text en Copyright © 2014 Justas Lazutka et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Lazutka, Justas
Zvirbliene, Aurelija
Dalgediene, Indre
Petraityte-Burneikiene, Rasa
Spakova, Aliona
Sereika, Vilimas
Lelesius, Raimundas
Wernike, Kerstin
Beer, Martin
Sasnauskas, Kestutis
Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
title Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
title_full Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
title_fullStr Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
title_full_unstemmed Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
title_short Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
title_sort generation of recombinant schmallenberg virus nucleocapsid protein in yeast and development of virus-specific monoclonal antibodies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4058894/
https://www.ncbi.nlm.nih.gov/pubmed/24982920
http://dx.doi.org/10.1155/2014/160316
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