葡聚糖接枝型Protein A介质的层析性能提升  

Binding Enhancement by Dextran-grafting to Protein A Agarose-based Affinity Medium

作　　者：朱凯[1,2] 赵岚[2] 黄永东[2] 李强[2] 邱晗[3] 王启宝[1] 苏志国[2] 马光辉[2,4] ZHU Kai ZHAO Lan HUANG Yong-dong LI Qiang QIU Han WANG Qi-bao SU Zhi-guo MA Guang-hui(School of Chemical and Environment Engineering, China University of Mining and Technology Beijing, Beijing 100083, China National Key Lab. Biochem. Eng., Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China Department of Applied Chemistry, Beijing Institute of Petrochemical Technology, Beijing 102617, China Jiangsu National Synergetic Innovation Center for Advanced Materials （SICAM）, Nanjing, Jiangsu 210011, China)

机构地区：[1]中国矿业大学(北京)化学与环境工程学院,北京100083 [2]中国科学院过程工程研究所生化工程国家重点实验室,北京100190 [3]北京石油化工学院应用化学系,北京102617 [4]国家江苏先进生物与化学制造协同创新中心,江苏南京210011

出　　处：《过程工程学报》2016年第5期856-861,共6页The Chinese Journal of Process Engineering

基　　金：国家自然科学基金资助项目(编号:213062026;21206175;21476241);国家科技支撑计划资助项目(编号:2013BAB01B03);国家重大仪器基金资助项目(编号:2013YQ14040502);国家高技术研究发展计划(863)基金资助项目(编号:2014AA021006);北京市大学生毕业论文提升基金资助项目(编号:BIPT-SPBYSJ201502)

摘　　要：Sepharose 4FF微球经环氧活化后与葡聚糖溶液反应,得葡聚糖接枝型琼脂糖微球,再经环氧活化和偶联耐碱型Protein A配基,得葡聚糖接枝型高载量Protein A介质,测定了介质在线清洗稳定性能,并进行了热力学研究.结果表明,与常规Protein A介质相比,葡聚糖接枝型Protein A介质的最高流速提高约32%,对抗体h Ig G的动态载量为60.6 mg/m L,分别为常规介质和Mab Select Su Re介质载量的123%和95%;经40次清洗后,葡聚糖接枝型Protein A介质动态载量为原始载量的92%,远高于常规介质的84%,与Mab Select Su Re稳定性基本一致.3种介质对抗体的结合均为熵驱动过程,葡聚糖接枝型Protein A介质的吸附热介于Mab Select Su Re和常规Protein A介质之间.After being activated by epoxy groups, Sepharose 4FF was grafted with dextran and then epoxy-activated followed by being coupled with alkali-resistant protein A ligand. In this paper, the cleaning-in-place performance of the medium is measured, and thermodynamic studies were carried out. Compared with non-grafted medium, the maximum velocity of dextran-grafted protein A medium was increased by about 32%. The binding capacity of antibody hIgG of dextran-grafted protein A medium was 60.6 mg/mL, which has reached 123% and 95% of the values for non-grafted protein A and MabSelect SuRe. After 40 cleaning-in-place cycles rinsed, the dynamic binding capacity of dextran-grafted protein A maintained the original capacity of 92%, which was almost the same as MabSelect SuRe and was also much higher than non-grafted Protein A. The capacity of non-grafted protein A maintained the original capacity of 84%. Thermodynamic studies showed that the binding processes of three chromatographic media were entropy-driven processes. The adsorption heat of dextran-grafted protein A medium was between that of MabSelect SuRe and non-grafted protein A.

关 键 词：抗体纯化 葡聚糖接枝 PROTEIN A层析 载量 

分 类 号：Q503[生物学—生物化学]