疏水性电荷诱导色谱分离抗HER2单克隆抗体

doi:10.3969/j.issn.0438-1157.2014.10.025

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

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 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

收稿日期:2014-01-27 修回日期:2014-03-18 出版日期:2014-10-05 发布日期:2014-10-05
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）。

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 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).

摘要/Abstract

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

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

中图分类号:

TQ028.8

李菁, 林东强, 童红飞, 姚善泾, 叶帅东, 崔加友. 疏水性电荷诱导色谱分离抗HER2单克隆抗体[J]. 化工学报, 2014, 65(10): 3931-3937.

LI Jing, LIN Dongqiang, TONG Hongfei, YAO Shanjing, YE Shuaidong, CUI Jiayou. Separation and purification of anti-HER2 monoclonal antibody with hydrophobic charge-induction chromatography[J]. CIESC Journal, 2014, 65(10): 3931-3937.

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疏水性电荷诱导色谱分离抗HER2单克隆抗体

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

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