CAJ | 학술논문

目的大量制备、纯化抗人G250单克隆抗体(G250-MeAb),并鉴定其与G250合成多肽及细胞天然抗原的结合特异性.方法将抗人G250杂交瘤细胞接种BALB/C小鼠腹腔,大量制备G250-McAb.采用正辛酸-硫酸铵沉淀法获得粗制抗体,然后经蛋白A亲和层析柱纯化抗体.采用荧光激活细胞分类术及免疫组织化学法鉴定G250-McAb与抗原结合特异性.结果成功制备抗人G250-MeAb,腹水经纯化后,获得了高纯度抗人G250单克隆抗体,蛋白浓度达4.78 g/L.荧光激活细胞分类术及免疫组织化学结果显示G250-McAb与G250表达阳性的Ketr-3细胞特异性结合,而不与G250表达阴性的ACHN细胞结合.结论成功制备并纯化G250-McAb,为进一步开展G250-McAb介导的肾癌诊断、治疗奠定了基础.
목적 대량제비、순화항인G250단극륭항체(G250-MeAb),병감정기여G250합성다태급세포천연항원적결합특이성.방법 장항인G250잡교류세포접충BALB/C소서복강,대량제비G250-McAb.채용정신산-류산안침정법획득조제항체,연후경단백A친화층석주순화항체.채용형광격활세포분류술급면역조직화학법감정G250-McAb여항원결합특이성.결과 성공제비항인G250-MeAb,복수경순화후,획득료고순도항인G250단극륭항체,단백농도체4.78 g/L.형광격활세포분류술급면역조직화학결과 현시G250-McAb여G250표체양성적Ketr-3세포특이성결합,이불여G250표체음성적ACHN세포결합.결론 성공제비병순화G250-McAb,위진일보개전G250-McAb개도적신암진단、치료전정료기출.
Objective To prepare and purify anti-human-G250 monoclonal antibody (G250-McAb) and identify its function. Methods G250-McAb was generated in BALB/C mice abdominal cavity by injecting intraperitoneally with 1 × 106 hybridoma cells. The purity of McAb was determined by ammonium sulphate. The specificity of the McAb with G250 antigen was identified by fluorescence-activated cell sorting (FACS) and immunohischemistry. Results The McAb with high purity was obtained and the final concentration of McAb protein was about 4.78 g/L. It was also verified by FACS and immunohischemistry that McAb could bind to the surface of the G250 antigen positively expressed cells such as Ketr-3 but not to the G250 antigen negatively expressed cells such as ACHN. Conclusion Anti-human-G250 McAb with high specificity was successfully generated and evaluated,which can be used in the diagnosis and treatment of renal cancer.