Page 7 - 南京医科大学自然版

第44卷第6期 南京医科大学学报（自然科学版）
2024年6月 Journal of Nanjing Medical University（Natural Sciences） ·743 ·

·基础研究·

全人源抗大别班达病毒抗体的制备及功能鉴定

徐若薇 ，韩一芳 ，周婷婷 ，吴梓菡 ，李 军 ，金 柯 ，汪春晖 1，2*
2
2
3
2
1
3
南京师范大学生命科学学院，江苏 南京 210046；东部战区疾病预防控制中心传染病防控科，江苏 南京 210018；南京
1 2 3
医科大学第一附属医院感染科，江苏 南京 210029
［摘 要］ 目的：从发热伴血小板减少综合征（severe fever with thrombocytopenia syndrome，SFTS）恢复期患者的外周血单核细
胞（peripheral blood mononuclear cell，PBMC）中，获得大别班达病毒（Dabie bandavirus，DBV）特异性抗体序列，制备全人源DBV
抗体。方法：用酶联免疫吸附试验（enzyme⁃linked immunosorbent assay，ELISA）检测恢复期患者血浆抗体效价，挑取恢复期患
者PBMC样本进行磁珠富集，经流式细胞仪分选获得能够结合DBV糖蛋白的特异性单个B细胞，经B细胞受体（B cell receptor，
BCR）测序获得抗体序列，进行单链抗体的原核表达，采用蛋白免疫印迹、ELISA和体外中和实验鉴定抗体的表达、结合能力和
中和活性。结果：共收集25例SFTS恢复期患者血液样本，血浆中均发现了DBV糖蛋白和核衣壳蛋白的抗体。选取样本进行
流式分选和BCR测序，分析后共获得6个单链抗体，对DBV均具有良好的结合能力和中和活性。结论：成功表达了6个与DBV
具有中和作用的全人源抗DBV抗体。
［关键词］ 发热伴血小板减少综合征；大别班达病毒；恢复期患者；全人源抗体
［中图分类号］ R813.2 ［文献标志码］ A ［文章编号］ 1007⁃4368（2024）06⁃743⁃10
doi：10.7655/NYDXBNSN240190

Preparation and functional identification of fully human antibodies against Dabie bandavirus

3
2
3
2
1
2
XU Ruowei ，HAN Yifang ，ZHOU Tingting ，WU Zihan ，LI Jun ，JIN Ke ，WANG Chunhui 1，2*
1 School of Life Sciences，Nanjing Normal University，Nanjing 210046；Infectious Disease Prevention and Control
2
3
Division，Eastern Theater Disease Prevention and Control Center，Nanjing 210018；Infectious Diseases Department，
the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，China
［Abstract］ Objective：To obtain Dabie bandavirus（DBV）specific antibody sequences from peripheral blood mononuclear cell
（PBMC）of recovery patients with severe fever with thrombocytopenia syndrome（SFTS）. Then fully human antibodies against DBV
were prepared. Methods：Enzyme⁃linked immunosorbent assay（ELISA）was used to detect the plasma antibody of SFTS recovery
patients. PBMC samples from the recovery patients were selected for magnetic bead enrichment, and the specific single B cells with
DBV glycoprotein（DBV⁃Gn）protein binding activity were obtained by flow cytometry. The antibody sequences were obtained by B cell
receptor sequencing，and the prokaryotic expression of single chain antibody was performed. Finally，the expression，binding capacity，
and neutralizing activity of the antibodies were identified by Western blot，ELISA，and neutralization experiment in vitro. Results：Total
25 blood samples from SFTS recovery patients were collected. The antibodies showed binding activity with DBV ⁃ Gn and DBV
nucleocapsid protein（DBV⁃NP）were detected in all plasma. Six types of single chain antibodies were obtained after flow cytometry
sorting and BCR sequencing analysis，all of which had good binding capacity and neutralizing activity with DBV. Conclusion：Six fully
human antibodies with neutralizing activity to DBV have been successfully expressed.
［Key words］ severe fever with thrombocytopenia syndrome；Dabie bandavirus；recovery patient；fully human antibody
［J Nanjing Med Univ，2024，44（06）：743⁃752］

［基金项目］ 国家自然科学基金（81871242）；“逸启”自主科
发热伴血小板减少综合征（severe fever with
研基金（2024YQZL01，2023YQFH05）；军队后勤科研计划项
目（BLB23J017G） thrombocytopenia syndrome，SFTS）是近年来在我国
［1］
∗ 部分地区发现的一种自然疫源性疾病 ，病原体为
通信作者（Correspondingauthor），E⁃mail：13912966353@139.com