Protein expression and antibody preparation of the nucleoprotein of sever fever with thrombocytopenia syndrome virus
Lu Huabao
Cheng Ziwen
Hu Yingfang
Wang Junzhu
Zhang Xiaoru
Sun Huidi
Wang Dawei
Yang Baoling
Abstract：This study aimed to express recombinant severe fever with thrombocytopenia syndrome virus(SFTSV)nucleoprotein(NP)and immunize mice to prepare polyclonal antibodies,laying the foundation for the development of SFTSV diagnostic reagents.The NP gene of SFTSV was amplified and cloned into the pET-28a prokaryotic expression plasmid.Protein expression and purification were performed under optimized conditions.Mice were immunized to generate specific antibodies,and the antibody titer was evaluated by ELISA.The specificity of the antibodies was further analyzed using Western blot and immunofluorescence assay(IFA).Results showed that the 738 bp SFTSV-NP gene was successfully amplified and ligated into the pET-28a plasmid,as confirmed by colony PCR and sequencing.By optimizing expression conditions,maximum protein yield(27 kDa)was achieved at 28℃with 0.1 mmol/L IPTG induction.Large-scale expression and purification were subsequently performed.ELISA confirmed that the antibody titer met experimental requirements,while Western blot and IFA demonstrated excellent specificity.This study successfully purified SFTSV-NP protein and prepared polyclonal antibodies,providing a basis for further research on SFTSV surveillance and functional studies.
Keywords：Severe fever with thrombocytopenia syndrome virus(SFTSV)NucleoproteinProkaryotic expressionPolyclonal antibodies
Publication Date：2025-03-15
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 26-31 )