摘要
为了研制骨桥蛋白(OPN)特异性单克隆抗体(mAb),并鉴定其特异性。本研究在毕赤酵母中表达具有良好免疫原性的重组人OPN蛋白的基础上,用重组人骨桥蛋白(rhOPN)免疫BALB/c小鼠,取其脾细胞与小鼠骨髓瘤细胞融合,间接ELISA筛选杂交瘤细胞,并结合免疫印迹对抗体的特异性进行鉴定,通过竞争抑制试验对单克隆抗体识别的抗原位点进行分析。结果共获得4株能够识别OPN不同抗原位点的mAb,亚类测定显示,3株为IgG1,1株为IgG2a。这些mAb能与重组人OPN特异性结合。本研究的四株抗体中,只有4G2B5能够检出与肿瘤转移密切相关的OPN-c的条带,预示该抗体可用于判断肿瘤预后和高转移肿瘤的临床检测。本研究成功获得了针对骨桥蛋白的特异性mAb,同时为进一步研究OPN蛋白的结构和功能提供了重要工具。
BALB/c mice were immunized with recombinant human osteopontin(rhOPN) and splenocytes of the immunized mice were fused with myeloma NS1 cells to produce hybridoma cell lines.ELISA assay was used to identify the monoclonal antibody against rhOPN protein and Western blotting was applied to identify the specificity of this antibody.It was demonstrated that 4 monoclonal antibodies(mAbs) recognizing different epitopes on OPN were obtained.These antibodies showed specific binding activity to recombinant OPN protei...
出处
《现代免疫学》
CAS
CSCD
北大核心
2008年第6期474-478,共5页
Current Immunology
关键词
骨桥蛋白
肿瘤侵袭
单克隆抗体
osteopontin
invasive tumor
monoclonal antibodies
