Identification and characterization of human monoclonal antibodies against the nuclear protein of severe fever with thrombocytopenia syndrome virus
10.3760/cma.j.cn112866-20240927-00144
- VernacularTitle:严重发热伴血小板减少综合征病毒核蛋白全人源单克隆抗体筛选和鉴定
- Author:
Binyang ZHENG
1
;
Zhifeng LI
;
Chuankun YANG
;
Hongxing PAN
;
Li ZHANG
Author Information
1. 江苏省疾病预防控制中心 江苏省新发突发重大传染病病原微生物重点实验室，南京 210009
- Publication Type:Journal Article
- Keywords:
Severe fever with thrombocytopenia syndrome virus;
Nuclear protein;
Phage display;
Human monoclonal antibody
- From:
Chinese Journal of Experimental and Clinical Virology
2024;38(6):694-701
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To screen human monoclonal antibodies (mAbs) against the nuclear protein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV), identify their binding specificity to both recombinant NP and NP in viral particles, and determine their affinity constant and binding kinetics.Methods:Antibody genes were extracted from the blood of recovered individuals, and an antibody library was created using phage display. This library was panned by recombinant NP. The selected antibodies were expressed and purified. Enzyme linked immunosorbent assay (ELISA), western blot (WB), and indirect immunofluorescence assay (IFA) were used to assess the binding specificity of these mAbs to recombinant NP and NP in virions. Additionally, biolayer interferometry (BLI) was utilized to determine the antibody affinity constant.Results:An antibody library with a capacity of 7.24×10 7 was successfully constructed. Following three rounds of panning, 6 mAbs (named as NP-1, NP-10, NP-11, NP-20, NP-21, and NP-27) were isolated. The binding specificity of these 6 mAbs against recombinant NP was confirmed through indirect ELISA and WB analysis. Additionally, these mAbs were demonstrated specific in binding to NP in virions as evidenced by IFA detection. The affinity constant values of the 6 mAbs, determined by BLI assay, ranged from 0.47 nmol/L to 32 nmol/L. Conclusions:The 6 mAbs derived from the library are human mAbs that exhibit specificity to the NP of SFTSV and demonstrate a high affinity. These antibodies represent potential candidates for fundamental research and development of diagnostic reagents for SFTSV.
