摘要
对6种商品化蛋白A亲和层析介质进行了性能分析,包括动态载量、洗脱条件等,并对含有单抗药物Mab-HS006的实际料液进行了捕获分离,对宿主细胞蛋白及蛋白A的残留进行了分析。结果表明,Prosep Ultra Plus和Mabselect SuRe亲和介质的载量最高,而Protein A Sepharose CL-4B和r-Protein A Sepharose的洗脱条件最温和。一步层析捕获分离Mab-HS006单抗的结果表明,各种介质的纯度均可达90%以上,Prosep Ultra Plus的宿主细胞蛋白残留最高,Mabselect SuRe蛋白A的脱落最低。
During the selection of protein A the affinity media for the purification of monoclonal antibodies,dynamic binding capacity,elution conditions and purity are essential parameters to be evaluated.In this study,these performances were conducted for six kinds of affinity media.Furthermore,the media were used for purification of monoclonal antibody Mab-HS006 from crude CHO fermentation supernatant.A larger dynamic binding capacity was found for Prosep Ultra Plus and Mabselect SuRe,and a wilder elution condition was found for Protein A Sepharose CL-4B and r-Protein A Sepharose.All the media could achieve a high purity of over 90% and recovery of more than 98%.For the media Prosep Ultra Plus,the highest host cell protein content of eluates was found,whereas for Mabselect SuRe the lowest protein A leakage level was achieved.
出处
《药物生物技术》
CAS
CSCD
2012年第1期40-44,共5页
Pharmaceutical Biotechnology
基金
中国博士后科学基金资助项目(20110491826)
关键词
蛋白A亲和层析
动态载量
洗脱行为
单克隆抗体
纯化
Protein A affinity chromatography
Dynamic binding capacity
Elution conditions
Monoclonal antibody
Purification