作　　者： ; ; ; ; ; ; ; ;

机构地区： 中国矿业大学北京化学与环境工程学院

出　　处： 《过程工程学报》 2016年第5期856-861,共6页

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 层析 载量

领　　域： [生物学]