title: Generation of recombinant Schmallenberg virus nucleocapsid protein in yeast and development of virus-specific monoclonal antibodies.
abstract: 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.
authors: Institute of Biotechnology, Vilnius University, V.A. Graiciuno 8, 02241 Vilnius, Lithuania.
authors: Justas
authors: 0000-0003-2006-7280
authors: J
authors: Lazutka
authors: Institute of Biotechnology, Vilnius University, V.A. Graiciuno 8, 02241 Vilnius, Lithuania.
authors: Aurelija
authors: A
authors: Zvirbliene
authors: Institute of Biotechnology, Vilnius University, V.A. Graiciuno 8, 02241 Vilnius, Lithuania.
authors: Indre
authors: I
authors: Dalgediene
authors: Institute of Biotechnology, Vilnius University, V.A. Graiciuno 8, 02241 Vilnius, Lithuania.
authors: Rasa
authors: 0000-0001-5676-2993
authors: R
authors: Petraityte-Burneikiene
authors: Institute of Biotechnology, Vilnius University, V.A. Graiciuno 8, 02241 Vilnius, Lithuania.
authors: Aliona
authors: A
authors: Spakova
authors: Institute of Microbiology and Virology, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes 18, 47181 Kaunas, Lithuania.
authors: Vilimas
authors: V
authors: Sereika
authors: Institute of Microbiology and Virology, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes 18, 47181 Kaunas, Lithuania.
authors: Raimundas
authors: R
authors: Lelesius
authors: Institut für Virusdiagnostik, Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany.
authors: Kerstin
authors: 0000-0001-8071-0827
authors: K
authors: Wernike
authors: Institut für Virusdiagnostik, Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany.
authors: Martin
authors: M
authors: Beer
authors: Institute of Biotechnology, Vilnius University, V.A. Graiciuno 8, 02241 Vilnius, Lithuania.
authors: Kestutis
authors: 0000-0002-8770-0645
authors: K
authors: Sasnauskas
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis; D002044:Bunyaviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000276:immunology / Q000662:veterinary*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000276:immunology / Q000821:virology; D002460:Cell Line; D006224:Cricetinae; D005260:Female; D015870:Gene Expression; D007106:Immune Sera; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis; D008097:Lithuania / Q000453:epidemiology; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D016851:Orthobunyavirus / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007106:Immune Sera; D007074:Immunoglobulin G; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins