遵义医科大学学报2026，Vol.49Issue(8)：840-847,8.
人博卡病毒1型特异性单克隆抗体的制备及其抗原检测初步应用
Development of a HBoV1 specific monoclonal antibody 5A1 and its preliminary application in antigen detection
摘要
Abstract
Objective To develop a genotype specific monoclonal antibody(mAb)targeting the VP3 DR2 re-gion of human bocavirus 1(HBoV1)and systematically characterize its specificity,affinity,and antigen detec-tion efficacy.Methods The DR2 peptide of HBoV1 was used as antigen to immunize BALB/c mice for the selec-tion of hybridoma cell lines stably secreting high titer anti-HBoV1-DR2 mAb.The genotype specificity of the generated mAb were assessed using enzyme-linked immunosorbent assay(ELISA),indirect immunofluorescence assay(IFA),and bio-layer interferometry(BLI)with the affinity constant(KD).The structure of the mAb and its interaction with HBoV1 DR2 were predicted using AlphaFold2 and molecular docking.The clinical validity was evaluated among clinical specimens.Results One anti-HBoV1-DR2 mAb,designated as 5A1 and identified as IgG1/κ subtype,was successfully generated.The titer of 5A1 binding to HBoV1 VP3(1:25 600)is signifi-cantly higher than that to HBoV2 VP3(<1:400).Specific green fluorescence signal was only observed in HEK293 cells transfected with pcDNA3.1-HBoV1 VP3.The KD value of 2.21×10-11(M)in BLI revealed a high affinity of 5A1 with HBoV1 VP3.Molecular docking predicted hydrogen bonds between LEU22/ALA13 of peptide DR2 and LYS-43A of the 5A1 Fab,and ASP-9 of peptide DR2 and LYS-107B of the 5A1 Fab.The me-dian viral load(7.33×107copies/ml)in10 antigen-positive specimens was significantly higher than that(3.88×104copies/ml)in 23 antigen-negative specimens(P=0.000 5).Conclusion A genotype specific mAb,5A1,was harvested in the study,which showed high specificity and affinity with HBoV1 VP3,but not HBoV2 VP3,and high validity in antigen test among clinical specimens.关键词
人博卡病毒1 型/型别特异性/VP3 蛋白DR2 区/单克隆抗体/亲和力Key words
human bocavirus 1/monoclonal antibody/molecular docking/antigen detection分类
医药卫生引用本文复制引用
德日,傅涵浩钰,张可祥,孙宇,贾立平,赵林清..人博卡病毒1型特异性单克隆抗体的制备及其抗原检测初步应用[J].遵义医科大学学报,2026,49(8):840-847,8.基金项目
国家自然科学基金资助项目(NO:82502717) （NO:82502717）
北京市自然科学基金资助项目(NO:7254305) （NO:7254305）
国家自然科学基金面上项目(NO:82172277). （NO:82172277）
