CAJ | 학술논문

目的表达纯化人博卡病毒( human Bocavirus,HBoV) VP2蛋白,采用杂交瘤方法制备抗HBoV VP2蛋白的单克隆抗体.方法应用原核表达载体pET-30a和大肠埃希菌表达VP2蛋白,并用固定化金属亲和层析,纯化后的蛋白作为抗原免疫BALB/c小鼠,利用杂交瘤技术和间接ELASA方法筛选阳性的杂交瘤细胞,并对单克隆抗体进行分型检测和滴度测定.结果表达并纯化获得了重组HBoV VP2蛋白,利用杂交瘤技术得到单克隆IgG抗体,抗体效价达到1∶4×105.结论利用HBoV VP2蛋白免疫制备了单克隆抗体,并具有较高的效价.本研究为快速诊断和研究HBoV打下基础.
목적 표체순화인박잡병독( human Bocavirus,HBoV) VP2단백,채용잡교류방법제비항HBoV VP2단백적단극륭항체.방법 응용원핵표체재체pET-30a화대장애희균표체VP2단백,병용고정화금속친화층석,순화후적단백작위항원면역BALB/c소서,이용잡교류기술화간접ELASA방법사선양성적잡교류세포,병대단극륭항체진행분형검측화적도측정.결과 표체병순화획득료중조HBoV VP2단백,이용잡교류기술득도단극륭IgG항체,항체효개체도1∶4×105.결론 이용HBoV VP2단백면역제비료단극륭항체,병구유교고적효개.본연구위쾌속진단화연구HBoV타하기출.
Objective To express and purify HBoV VP2 protein,and the monoclonal antibody against HBoV VP2 protein was prepared with hybridoma technique.Methods The HBoV VP2 cloned into vector pET-30a was expressed in E.coil.After purified by immobilized metal affinity chromatography,the BALB/c mouse was immunized with purified protein as antigen.The positive hybridoma cells were screened with hybridoma technique and ELISA assay. Isotype and titer of the monoclonal antibody were detected.Results The recombinant HBoV VP2 protein was expressed and purified,and then the monoclonal antibody was obtained with hybridoma technique.The titer of the IgG monoclonal antibody was up to 1 ∶ 4 × 105.Conclusion Monoclonal antibody against recombinant HBoV VP2 protein was prepared and the antibody titer was high.This work may provide a new method in rapid diagnosis and study of HBoV.