Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies
Autor: | Aurelija Zvirbliene, Kestutis Sasnauskas, Vilimas Sereika, Justas Lazutka, Kerstin Wernike, Raimundas Lelešius, Indre Dalgediene, Martin Beer, Rasa Petraityte-Burneikiene, Aliona Špakova |
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Jazyk: | angličtina |
Rok vydání: | 2014 |
Předmět: |
lcsh:Immunologic diseases. Allergy
Orthobunyavirus Article Subject medicine.drug_class Immunology Cattle Diseases Gene Expression Saccharomyces cerevisiae Immunofluorescence Monoclonal antibody Antibodies Viral Bunyaviridae Infections Virus Serology law.invention Cell Line Mice law Cricetinae medicine Immunology and Allergy Animals Mice Inbred BALB C medicine.diagnostic_test biology Immune Sera Antibodies Monoclonal Schmallenberg virus Lithuania General Medicine Nucleocapsid Proteins biology.organism_classification Virology Recombinant Proteins Immunoassay Immunoglobulin G biology.protein Recombinant DNA Cattle Female Immunization Antibody lcsh:RC581-607 Research Article |
Zdroj: | Journal of Immunology Research, Vol 2014 (2014) Journal of Immunology Research |
ISSN: | 2314-7156 2314-8861 |
Popis: | 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. |
Databáze: | OpenAIRE |
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