Characterization of high-affinity antibodies against the surface Gc protein of Dabie bandavirus / severe fever with thrombocytopenia syndrome virus

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemistry and Biophysics Reports Pub Date : 2024-07-09 DOI:10.1016/j.bbrep.2024.101779
Pyeonghwa Jeon , Bin Yoo , Yoonji Kim , So-Young Lee , Hye-Min Woo , Hee-Young Lim , Joo-Yeon Lee , Sora Park , Hansaem Lee
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Abstract

Severe fever with thrombocytopenia syndrome virus (SFTSV) or Dabie bandavirus is an emerging pathogen responsible for SFTS. It is considered a novel threat to human health, given the high associated fatality. SFTSV is a segmented negative-strand RNA virus containing three single-stranded RNAs, with the M segment encoding the glycoproteins Gn and Gc. Gc is vital for viral entry into the host cell surface, along with the Gn protein. As the Gc is the surface-exposable antigen from virions, it is a critical diagnostic marker of infection. Although various SFTSV Gn or N protein-based sero-diagnostic methods have been developed, there are no commercially available sero-diagnostic kits. Therefore, we generated monoclonal antibodies (mAbs) against SFTSV Gc and explored their application in serum diagnostic tests to develop sensitive serodiagnostic tools covering broad-range genotypes (A to F). First, 10 SFTSV Gc antibody-binding fragments (Fabs) were isolated using a phage display system and converted into human IgGs. Enzyme-linked immunosorbent assays (ELISA) of the SFTSV and Rift Valley fever virus (RVFV: same genus as SFTSV) Gc antigens showed that all antibodies attached to the SFTSV Gc protein had high affinity. An immunofluorescence assay (IFA), to verify the cross-reactivity of seven antibodies with high affinities for various SFTSV genotypes (A, B2, B3, D, and F) and detect mAb binding with intact Gc proteins, revealed that five IgG type mAbs were bound to intact Gc proteins of various genotypes. Six high-affinity antibodies were selected using ELISA and IFA. The binding capacity of the six antibodies against the SFTSV Gc antigen was measured using surface plasmon resonance. All antibodies had high binding capacity. Consequently, these antibodies serve as valuable markers in the serological diagnosis of SFTSV.

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针对达比带状疱疹病毒/严重发热伴血小板减少综合征病毒表面Gc蛋白的高亲和力抗体的特征描述
严重发热伴血小板减少综合征病毒(SFTSV)或达比带状疱疹病毒是一种导致严重发热伴血小板减少综合征的新病原体。由于其致死率很高,因此被认为是对人类健康的一种新威胁。SFTSV 是一种分段负链 RNA 病毒,含有三条单链 RNA,其中 M 段编码糖蛋白 Gn 和 Gc。Gc 与 Gn 蛋白一起对病毒进入宿主细胞表面至关重要。由于 Gc 是病毒表面可暴露的抗原,因此是感染的重要诊断标志。虽然目前已开发出多种基于 SFTSV Gn 或 N 蛋白的血清诊断方法,但还没有商业化的血清诊断试剂盒。因此,我们生成了针对 SFTSV Gc 的单克隆抗体(mAbs),并探索了它们在血清诊断测试中的应用,以开发出涵盖广泛基因型(A 至 F)的灵敏血清诊断工具。首先,利用噬菌体展示系统分离出 10 个 SFTSV Gc 抗体结合片段(Fabs),并将其转化为人类 IgG。SFTSV和裂谷热病毒(RVFV:与SFTSV同属)Gc抗原的酶联免疫吸附试验(ELISA)显示,所有与SFTSV Gc蛋白相连的抗体都具有很高的亲和力。免疫荧光试验(IFA)验证了七种对不同 SFTSV 基因型(A、B2、B3、D 和 F)具有高亲和力的抗体的交叉反应性,并检测了 mAb 与完整 Gc 蛋白的结合情况,结果显示有五种 IgG 型 mAb 与不同基因型的完整 Gc 蛋白结合。利用 ELISA 和 IFA 方法选出了六种高亲和力抗体。利用表面等离子共振测量了这六种抗体与SFTSV Gc抗原的结合能力。所有抗体都有很高的结合能力。因此,这些抗体可作为SFTSV血清学诊断的重要标志物。
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来源期刊
Biochemistry and Biophysics Reports
Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics
CiteScore
4.60
自引率
0.00%
发文量
191
审稿时长
59 days
期刊介绍: Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.
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