CIESC Journal ›› 2014, Vol. 65 ›› Issue (10): 3931-3937.DOI: 10.3969/j.issn.0438-1157.2014.10.025

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Separation and purification of anti-HER2 monoclonal antibody with hydrophobic charge-induction chromatography

LI Jing1, LIN Dongqiang1, TONG Hongfei1, YAO Shanjing1, YE Shuaidong2, CUI Jiayou2   

  1. 1 Key Laboratory of Biomass Chemical Engineering, Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, Zhejiang, China;
    2 Amprotein Biological Products Research Ltd., Hangzhou 310018, Zhejiang, China
  • Received:2014-01-27 Revised:2014-03-18 Online:2014-10-05 Published:2014-10-05
  • Supported by:

    supported by the National Natural Science Foundation of China (21036005, 21276228)and Zhejiang Provincial Natural Science Foundation for Distinguished Young Scientists(LR12B06003).

疏水性电荷诱导色谱分离抗HER2单克隆抗体

李菁1, 林东强1, 童红飞1, 姚善泾1, 叶帅东2, 崔加友2   

  1. 1 生物质化工教育部重点实验室, 浙江大学化学工程与生物工程学系, 浙江 杭州 310027;
    2 杭州安瑞普生物制品研究有限公司, 浙江 杭州 310018
  • 通讯作者: 林东强
  • 基金资助:

    国家自然科学基金项目(21036005,21276228);浙江省杰出青年科学基金项目(LR12B06003)。

Abstract: Hydrophobic charge-induction chromatography (HCIC) is a new technology for biomolecule separation. In the present work HCIC was used to separate monoclonal antibody (mAb) anti-HER2 from Chinese Hamster ovary (CHO) cell culture broth with typical HCIC resin, MEP HyperCel. The stability of anti-HER2 mAb in cell culture broth was investigated firstly, and the adsorption of anti-HER2 mAb onto MEP HyperCel at different pH was compared. The adsorption capacities were high at the range of pH 6-9, but decreased significantly under acidic condition. The dynamic binding capacity of anti-HER2 mAb was determined with a packed bed, and the loading and elution conditions were optimized. The suitable separation conditions were obtained, and the purity of mAb could reach 94.6% with the yield of 0.1 mg·(ml broth)-1. It is feasible to separate mAb from mammalian cell culture broth with HCIC.

Key words: bioseparation, monoclonal antibody, chromatography, hydrophobic charge-induction chromatography, anti-HER2

摘要: 采用疏水性电荷诱导色谱(HCIC)新型生物分离方法,从CHO细胞培养液中高效分离抗HER2单克隆抗体(单抗)。选用典型HCIC介质MEP HyperCel,考察了细胞培养液中单抗稳定性,比较了不同pH下MEP HyperCel介质对抗HER2单抗的吸附性能,发现pH6~9范围内吸附容量均较高,酸性条件下吸附容量显著下降。进一步考察了抗HER2单抗的动态吸附载量,优化了上样和洗脱条件,确定了合适的分离条件,实现了细胞培养液中高效分离单抗,纯度达到94.6%,收率约0.1 mg·(ml料液)-1。结果表明,HCIC从哺乳动物细胞培养液中分离单抗是切实可行的,为单抗分离提供了新思路。

关键词: 生物分离, 单克隆抗体, 色谱, 疏水性电荷诱导色谱, 抗HER2

CLC Number: