抗膀胱肿瘤抗原单克隆抗体纯化方法的比较  

Comparison of Methods for Purification of Monoclonal Antibodies Against Bladder Tumor Antigens

作　　者：徐瑶瑶 朱文武 李洁 李净 王荣鹤 朱雄伟 XU Yaoyao;ZHU Wenwu;LI Jie;LI Jing;WANG Ronghe;ZHU Xiongwei(School of Environmental Ecology and Bioengineering,Wuhan Institute of Technology,Wuhan 430205,China;Wuhan Yanda Biotechnology Co.,Ltd,Wuhan 430200,China)

机构地区：[1]武汉工程大学环境生态与生物工程学院,湖北武汉430205 [2]武汉雁达生物技术有限公司,湖北武汉430200

出　　处：《武汉工程大学学报》2023年第1期76-80,共5页Journal of Wuhan Institute of Technology

基　　金：2021年湖北省大学生创新创业训练项目。

摘　　要：利用单克隆抗体制备,细胞融合及选择培养基筛选等技术制备抗膀胱肿瘤抗原单克隆抗体(BTA)。通过G蛋白亲和层析法(G柱法)和Q-琼脂糖亲和介质离子交换层析法(蓝胶加Q柱层析法)分别提取抗体,以SDS聚丙烯酰胺凝胶电泳、紫外分光光度计、酶联免疫吸附试验双抗体夹心法对两种不同方法提取的抗BTA抗体进行分析,发现G柱法得到的抗体含量明显高于蓝胶加Q柱法(P<0.05),抗体质量浓度达到60 ng/mL,G柱法提取的抗体与BTA的亲和力大于其他方法,且蓝胶加Q柱层析法生产样品活性为市售基准的97.11%(质量分数),G柱生产样品活性为101.55%(质量分数),这两种方法在抗BTA单克隆抗体纯化方面均可满足要求,可根据实际工艺状况及质控要求进行选择。The monoclonal antibodies against bladder tumor antigen were prepared using techniques such as monoclonal antibody preparation,cell fusion,and selective media screening. The monoclonal antibodies were extracted by G protein affinity chromatography(G-column chromatography) and Q-agarose affinity medium ion exchange chromatography(blue gum plus Q-column chromatography),respectively. The antibodies extracted by the two different methods were analyzed by sodium dodecyl sulfate polyacrylamide gelelectrophoresis electrophoresis, ultraviolet spectroscopy, and dual antibody sandwich enzyme-linked immunesorbent assay method. It was found that the antibody content obtained by the G-column method is significantly higher than that by the blue gum plus Q-column method(P<0.05),and after the antibody concentration reaches 60 ng/mL,the affinity between the antibody and the monoclonal antibodies against bladder tumor antigen extracted by the G-column method is greater than that by other methods. The sample activity produced by the blue gum plus Q-column chromatography method is 97.11% of the commercially available benchmark,while the G-column production sample activity is 101.55%. These two methods can meet the requirements in the purification of monoclonal antibodies against bladder tumor antigen,and can be selected according to the actual process conditions and quality control requirements.

关 键 词：膀胱肿瘤 抗体 离子交换层析法 亲和层析 

分 类 号：Q939.91[生物学—微生物学]