摘要
目的基于免疫组库测序筛选发热伴血小板减少综合征病毒(severe fever with thrombocytopenia syndrome virus,SFTSV)糖蛋白Gn单克隆抗体,并鉴定其生物学特性。方法以纯化的SFTSV-Gn作为诱饵,从免疫的雌性BALB/c小鼠B细胞库中钓取病毒特异性B细胞,利用10×免疫组库测序方法,结合生物信息学分析,挑选具有潜在结合活性的抗体序列,构建至真核表达载体pcDNA3.4,进行单克隆抗体的表达及纯化,并鉴定其生物学活性。结果从捕获的3544个B细胞中挑选出14株抗体,功能验证显示12株抗体能结合SFTSV-Gn糖蛋白,而其中C9和9/10抗体结合力较高;Western blot、间接免疫荧光试验(immunofluorescence assay,IFA)和流式细胞术证明该抗体能够特异性识别SFTSV,有效浓度低至2μg/mL。结论成功获得了针对SFTSV-Gn糖蛋白的单克隆抗体,为SFTSV快速诊断试剂的·开发奠定了基础。
Objective To obtain monoclonal antibodies against the glycoprotein Gn of severe fever with thrombocytopenia syndrome virus(SFTSV)based on immune repertoire sequencing and identify the biological characteristics.Methods Using purified SFTSV-Gn as bait,virus-specific B cells were captured from B cell library of immunized female BALB/c mice.Based on 10×immune repertoire sequencing and bioinformatics analysis,the antibody sequences with potential binding activity were selected and constructed into eukaryotic expression vector pcDNA3.4.The monoclonal antibodies were expressed and purified,and identified for the biological activity.Results Fourteen antibodies were selected from 3544 captured B cells,and the functional verification showed that 12 antibodies could bind to SFTSV-Gn glycoprotein,among which C9 and 9/10antibodies had higher binding ability.Western blot,indirect immunofluorescence assay(IFA)and flow cytometry proved that they specifically recognized SFTSV,and the effective concentration was as low as 2μg/mL.Conclusion The monoclonal antibodies against SFTSV-Gn glycoprotein were successfully obtained,which lays a foundation for the development of rapid diagnostic reagents for SFTSV.
作者
郑文君
张思伊
侯瑶
高艳洁
陈艳莉
朱庆祥
王梁
聂凯晓
于立娟
姬敬开
张振杰
全传松
ZHENG Wenjun;ZHANG Siyi;HOU Yao;GAO Yanjie;CHEN Yanli;ZHU Qingxiang;WANG Liang;NIE Kaixiao;YU Lijuan;JI Jingkai;ZHANG Zhenjie;QUAN Chuansong(Department of Pathogen Biology,School of Clinical and Basic Medical Sciences,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250014,Shandong Province,China;不详)
出处
《中国生物制品学杂志》
2025年第1期14-21,共8页
Chinese Journal of Biologicals
基金
山东省自然科学基金青年项目(ZR2020QH133)。
关键词
发热伴血小板减少综合征病毒
特异B细胞
单克隆抗体
Gn糖蛋白
Severe fever with thrombocytopenia syndrome virus(SFTSV)
Virus-specific B cells
Monoclonal antibodies
Gn glycoprotein
