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

• Published in: Applied microbiology and biotechnology Vol. 108; no. 1; p. 303
• Main Authors: Chang, Yu-Chih, Shimoda, Hiroshi, Jiang, Min-chao, Hsu, Yau-Heiu, Maeda, Ken, Yamada, Yumiko, Hsu, Wei-Li
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.12.2024; Springer Nature B.V
• Subjects: Antibodies; Applied Genetics and Molecular Biotechnology; Bamboo; bamboos; Biomedical and Life Sciences; Biotechnology; blood serum; Diagnosis; diagnostic techniques; Fever; Glycoproteins; Glycoproteins - metabolism; Huaiyangshan banyangvirus; Humans; Life Sciences; Microbial Genetics and Genomics; Microbiology; Nicotiana benthamiana; Phlebovirus - genetics; Phlebovirus - metabolism; Proteins; Purification; Sensitivity; Seroepidemiologic Studies; Serology; seroprevalence; Severe Fever with Thrombocytopenia Syndrome - diagnosis; Tags; Thrombocytopenia; Viruses; Severe fever with thrombocytopenia syndrome virus; SFTSV; Glycoprotein N (Gn); Glycosylation; based vector system; Secretory signal tags; Seroprevalence; Severe fever with thrombocytopenia syndrome virus (SFTSV); Bamboo mosaic virus-based vector system
• ISSN: 0175-7598; 1432-0614; 1432-0614
• DOI: 10.1007/s00253-024-13135-0

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%.