BTN3A3单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibody against BNTN3A3

作　　者：陆倩 柳迪 唐丽[1,2] LU Qian;LIU Di;TANG Li(Graduate School, Anhui Medical University, Hefei 230032, China;State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences · Beijing, Beijing Institute of Lifeomics, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 102206, China)

机构地区：[1]安徽医科大学研究生院,合肥230032 [2]军事科学院军事医学研究院生命组学研究所,蛋白质组学国家重点实验室,国家蛋白质科学中心(北京),北京102206

出　　处：《军事医学》2018年第2期105-109,共5页Military Medical Sciences

基　　金：国家自然科学基金资助项目(31570901)

摘　　要：目的制备BTN3A3单克隆抗体并鉴定。方法用含小鼠Ig G2a Fc标签的BTN3A3重组蛋白BTN3A3-m Fc作为抗原免疫BALB/c小鼠,通过细胞融合获得杂交瘤细胞。用含人Ig G1 Fc标签的BTN3A3重组蛋白BTN3A3-h Fc作为检测原,筛选分泌BTN3A3单抗的杂交瘤细胞株。利用Western印迹法和间接免疫荧光对上述单抗进行鉴定。结果共获得7株可稳定分泌BTN3A3单抗的杂交瘤细胞株。ELISA、Western印迹法和间接免疫荧光结果显示上述抗体可特异性地识别BTN3A3。结论获得可应用于ELISA、Western印迹法及免疫荧光的BTN3A3单抗,为BTN3A3的功能性研究奠定了基础。Objective To prepare and identify monoclonal antibodies against BTN3 A3. Methods BALB/c mice were immunized with the BTN3 A3 recombinant protein BTN3 A3-m Fc containing the mouse Ig G2 a Fc tag as an antigen,and hybridoma cells were obtained by cell fusion. BTN3 A3 recombinant protein BTN3 A3-h Fc containing a human Ig G1 Fc tag as a detection target was used to screen hybridoma cell lines secreting monoclonal antibodies against BTN3 A3. The above monoclonal antibodies were identified with Western blot and indirect immunofluorescence. Results Seven hybridoma cell lines that could stably secret BTN3 A3 monoclonal antibodies were obtained. ELISA, Western blot and indirect immunofluorescence showed that the above antibodies specifically recognized BTN3 A3. Conclusion BTN3 A3 monoclonal antibodies that can be applied to ELISA,Western blot or immunofluorescence have been obtained,which can contribute to the functional study of BTN3 A3.

关 键 词：嗜乳脂蛋白家族 BTN3A3 单克隆抗体 杂交瘤 免疫球蛋白类 

分 类 号：Q813.2[生物学—生物工程]