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

doi:10.3969/j.issn.0438-1157.2014.10.025

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 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 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

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:

TQ028.8

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.

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.

References 19

[1]	Yanga M, Clavijoa A, Grahama J, et al. Production and diagnostic application of monoclonal antibodies against influenza virus H5 [J]. Journal of Virological Methods, 2009, 162(1/2):194-202
[2]	Liu Meng(柳萌), Zhou Yali(周亚丽), Lao Xingzhen(劳兴珍), Zheng Heng(郑珩). Progress in study on purification of antibody by chromatography [J]. Pharmaceutical Biotechnology(药物生物技术), 2012, 19(2): 173-176
[3]	Wong N S, Anderson B O, Khoo K S, et a1. Breast cancer in developing countries [J]. Lancet, 2009, 374(9701): 1567
[4]	Moasser M M. Targeting the function of the HER2 oncogenc in human cancer therapeutics [J]. Oncogene, 2007, 26(46): 6577
[5]	Jiang Kuo. Advances in HER2 targeted therapy in breast cancer [J]. The Practical Journal of Cancer, 2010, 25(4): 425-432
[6]	Yao Shanjing(姚善泾), Gao Dong(高栋), Lin Dongqiang(林东强). Novel technology in bioseparation process-mixed-mode chromatography [J]. Journal of Chemical Industry and Engineering(China)(化工学报), 2007, 58(9): 2169-2177
[7]	Boschetti E. Antibody separation by hydrophobic charge induction chromatography [J]. Trends in Biotechnology, 2002, 20(8): 333-337
[8]	Chen Zhenming(陈振明), Lin Dongqiang(林东强), Yao Shanjing(姚善泾). New mixed-mode adsorbent and its application for separation of bovine colostrum immunoglobulin [J]. CIESC Journal (化工学报), 2012, 63(8): 2453-2459
[9]	Tong Hongfei(童红飞), Lin Dongqiang(林东强), Yao Shanjing(姚善泾). Separation and purification of immunoglobulin IgY with hydrophobic charge induction chromatography [J]. CIESC Journal (化工学报), 2011, 62(6): 1574-1580
[10]	Tong H F, Lin D Q, Yuan X M, Yao S J. Enhancing IgG purification from serum albumin containing feedstock with hydrophobic charge-induction chromatography [J]. Journal of Chromatography A, 2012, 1244: 116-122
[11]	Zhuang Tiantian(庄甜甜), Xia Haifeng(夏海峰), Lin Dongqiang(林东强),Yao Shanjing(姚善泾). Separation of immunoglobulin from porcine plasma with mixed-mode adsorption chromatography [J]. CIESC Journal (化工学报), 2010, 61(2): 336-341
[12]	Tong H F, Lin D Q, Pan Y, Yao S J. A new purification process for goose immunoglobulin IgY (Dfc) with hydrophobic charge-induction chromatography [J]. Biochemical Engineering Journal, 2011, 56(3): 205-211
[13]	Arakawa T, Kita Y, Sato H, Ejima D. MEP chromatography of antibody and Fc-fusion protein using aqueous arginine solution [J]. Protein Expression and Purification, 2009, 63(2): 158-163
[14]	Zhuang Tiantian(庄甜甜). Separation of immunoglobulin from porcine plasma with mixed-mode adsorption chromatography[D]. Hangzhou: Zhejiang University, 2010
[15]	Ghose S, Hubbard B, Cramer S M. Evaluation and comparison of alternatives to Protein A chromatography mimetic and hydrophobic charge induction chromatography stationary phases [J]. Journal of Chroamtograpyh A, 2006, 1122(1-2): 144-152
[16]	Chen Weidong(陈卫东), Sun Yan(孙彦). Effect of adsorption density on pore diffusivity of proteins in ion exchanger [J]. Journal of Chemical Industry and Engineering (China)(化工学报), 2003, 54(2):215-220
[17]	Lu H L, Lin D Q, Zhu M M, Yao S J. Effects of ligand density and pore size on the adsorption of bovine IgG with DEAE ion-exchange resins [J]. Journal of Separation Science, 2012, 35(16): 2131-2137
[18]	Ping W. Preparation of monoclonal antibody against human interleukin-15 and preliminary application[D]. Shijiazhuang: Hebei Medical University, 2003
[19]	Zhibing W. Optimizing cell culture conditions for Anti-CD3 mAbs and study of maximize productivity[D]. Shanghai: East China University of Science and Technology, 2011

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

疏水性电荷诱导色谱分离抗HER2单克隆抗体

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