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过程工程学报 ›› 2016, Vol. 16 ›› Issue (5): 856-861.DOI: 10.12034/j.issn.1009-606X.216151

• 生化工程专栏 • 上一篇    下一篇

葡聚糖接枝型耐碱Protein A介质的层析性能提升研究

朱凯1,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   

  1. 1. 中国矿业大学（北京）化学与环境工程学院
    2. 中国科学院过程工程研究所生化工程国家重点实验室
    3. 北京石油化工学院应用化学系
  • 收稿日期:2016-03-10 修回日期:2016-05-13 出版日期:2016-10-20 发布日期:2016-10-14
  • 通讯作者: 赵岚 lanzhao@ipe.ac.cn

ZHU Kai 1,ZHAO Lan 2,HUANG Yong-dong, 3,LI Qiang 4,QIU Han 5,WANG Qi-bao 6,SU Zhi-guo 7,MA Guang-hui 8   

  1. 1. China University of Mining and Technology, Beijing., School of Chemical and Environment Engineering
    2. State Key Lab.Biochem.Eng.,Institute of Process Engineering, Chinese Academy of Sciences
    3. State Key laboratory of Biochemical Engineering, Institute of Process Engineering, CAS
    4. National Key Lab of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences
    5. Department of Applied Chemistry, Beijing Institute of Petrochemical Technolog
    6. College of Chemical and Environmental Engineering,China University of Mining and Technology
    7. Key Laboratory of Biochemical Engineering，Institute of Process Engineering，Chinese Academy of Sciences
    8. State Key Lab. Biochem. Eng., Inst. Process Eng., Chinese Academy of Sciences
  • Received:2016-03-10 Revised:2016-05-13 Online:2016-10-20 Published:2016-10-14
  • Contact: ZHAO Lan lanzhao@ipe.ac.cn

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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hIgG的动态载量为60.6 mg/mL，分别为常规介质和MabSelect SuRe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MabSelect SuRe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MabSelect SuRe和常规Protein A介质之间.

关键词: 抗体纯化, 葡聚糖接枝, Protein A层析, 载量

Abstract: 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.

Key words: antibody purification, dextran-grafted, protein A chromatography, binding capacity