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Gn protein expressed in plants for diagnosis of severe fever with thrombocytopenia syndrome virus.

Yu-Chih ChangHiroshi ShimodaMin-Chao JiangYau-Heiu HsuKen MaedaYumiko YamadaWei-Li Hsu
Published in: Applied microbiology and biotechnology (2024)
Severe fever with thrombocytopenia syndrome virus (SFTSV) causes the highly fatal disease in humans. To facilitate diagnosis, the native form of subunit glycoprotein (Gn), a prime target for potential vaccines and therapies, was produced in Nicotiana benthamiana using a Bamboo mosaic virus-based vector system. By fusion with secretory signal tags, SS Ext , derived from the extension protein, and the (SP) 10 motif, the yield of the recombinant Gn (rGn) was remarkably increased to approximately 7 mg/kg infiltrated leaves. Ultimately, an rGn-based ELISA was successfully established for the detection of SFTSV-specific antibodies in serum samples from naturally infected monkeys. As validated with the reference method, the specificity and sensitivity of rGn-ELISA were 94% and 96%, respectively. In conclusion, utilizing well-suited fusion tags facilitates rGn production and purification in substantial quantities while preserving its antigenic properties. The rGn-ELISA, characterized by its commendable sensitivity and specificity could serve as a viable alternative diagnostic method for assessing SFTSV seroprevalence. KEY POINTS: • SFTSV Gn, fused with secretory signal tags, was expressed by the BaMV-based vector. • The plant fusion tags increased expression levels and eased the purification of rGn. • The rGn-ELISA was established and validated; its specificity and sensitivity > 94%.
Keyphrases
  • binding protein
  • monoclonal antibody
  • early onset
  • poor prognosis
  • amino acid
  • structural basis
  • protein protein
  • risk assessment
  • drug induced
  • long non coding rna
  • cell free