title: [Preparation and characterization of a monoclonal antibody against the nucleocapsid protein of Schmallenberg virus].
abstract: OBJECTIVE
  The present study was conducted to prepare and characterize a monoclonal antibody (mAb) against the nucleocapsid (N) protein of Schmallenberg virus (SBV).
  
  
  METHODS
  The SBV N gene was cloned into pET-28a-c(+ and pMAL-c5X vectors and then transformed into E.coli BL21. Histidine (His)-tagged (His-SBV-N) and maltose-binding protein (MBP)-tagged (MBP-SBV-N) fusion proteins were respectively induced to express by IPTG and purified by nickel-nitrilotriacetic acid (Ni-NTA) agarose and amylose resin. His-SBV-N was used to immunize BALB/c mice to prepare mAb, and MBP-SBV-N was used as the coating antigen in ELISA to screen mAb-secreting hybridomas and to determine mAb titers. The mAb against SBV N protein was purified from the ascitic fluids using protein G sepharose. Western blotting and indirect immunofluorescence assay were utilized to analyze the reactivity and specificity of the mAb.
  
  
  RESULTS
  One mAb specific for SBV N protein (named 1F2) was successfully screened and purified. The titer of 1F2 was 1:32 000. Besides, the isotype of 1F2 was determined to be IgG2α/κ. 1F2 reacted with both recombinant SBV N proteins and SBV isolates. It was also cross-reactive with the N proteins of genetically related Shamonda, Douglas and Akabane viruses, but not with the Rift Valley fever virus N protein.
  
  
  CONCLUSIONS
  One mAb specific for the SBV N protein was successfully prepared, it provides a useful tool for the serological detection of SBV.
authors.affiliation: Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China.
authors.forename: Yongning
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China.
authors.forename: Shaoqiang
authors.initials: S
authors.lastname: Wu
authors.affiliation: Friedrich-Loeffler-Institut, Greifswald 17493, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Wernike
authors.affiliation: Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China.
authors.forename: Jizhou
authors.initials: J
authors.lastname: Lv
authors.affiliation: Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Feng
authors.affiliation: Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China.
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Lin
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D003429:Cross Reactions; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D016851:Orthobunyavirus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins