组织因子单克隆抗体及对照抗体的制备、纯化和鉴定  

Purification and identification of tissue factor monoclonal antibodies

作　　者：亓翠玲[1] 郭四美[1] 叶杰[1] 周秦[1] 钟建国[1] 王丽京[1] 

机构地区：[1]广东药学院血管生物学研究所,广州510006

出　　处：《中国医疗前沿》2012年第11期24-25,共2页China Healthcare Innovation

基　　金：国家自然基金(编号:30900764);广东省医学科研基金(编号:A2011299)

摘　　要：目的纯化组织因子(TF)单克隆抗体及对照抗体。方法给小鼠腹腔注射杂交瘤8A3和9E10细胞,产生的腹水应用凝胶过滤层析法和Protein A亲和层析法纯化组织因子抗体及对照抗体,并用SDS-PAGE和BCA法检测纯化后单克隆抗体的纯度及蛋白浓度。结果单抗纯化后电泳重链和轻链都呈一条带,TF抗体和对照抗体的最终浓度分别为0.7726mg/ml和0.8592mg/ml。结论应用层析法和凝胶过滤和Protein A亲和分离层析法纯化抗体,能够得到纯度较高的单克隆抗体。Objective To purify and identify tissue factor monoclonal antibodies.Methods Firstly,8A3 and 9E10 cells were injected into abdominal cavity of BALB/c mice;then make affinity chromatography column with protein A-Sepharose CL-4B to purify tissue factor（TF） monoclonal antibodies.The purified specific TF monoclonal antibody was identificated and detected by SDS-PAGE and BCA.Results Purified TF antibody was electrophoresised and could show single band.The concentration of purified TF monoclonal antibody and control antibody respectively were 0.986mg/ml and 3.57mg/ml.Conclusion Affinity chromatography through protein A-Sepharose CL-4B can purify monoclonal antibody effectively.

关 键 词：组织因子 单克隆抗体 亲和层析 纯化 

分 类 号：R392.1[医药卫生—免疫学]