Experimental and in silico studies on three hydrophobic charge-induction adsorbents for porcine immunoglobulin purification

doi:10.1016/j.cjche.2015.06.019

Chinese Journal of Chemical Engineering ›› 2016, Vol. 24 ›› Issue (1): 151-157.DOI: 10.1016/j.cjche.2015.06.019

• 第25届中国过程控制会议专栏 • 上一篇下一篇

Experimental and in silico studies on three hydrophobic charge-induction adsorbents for porcine immunoglobulin purification

Qilei Zhang, Tiantian Zhuang, Hongfei Tong, Hongyin Wang, Dongqiang Lin, Shanjing Yao

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China

收稿日期:2014-10-13 修回日期:2015-05-06 出版日期:2016-01-28 发布日期:2016-02-23
通讯作者: Dongqiang Lin
基金资助:
Supported by the National Natural Science Foundation of China (21276228 and 21476198), the Natural Science Foundation of Zhejiang Province (LR12B06003), and the Fundamental Research Funds for the Central Universities (2013QNA4032).

Experimental and in silico studies on three hydrophobic charge-induction adsorbents for porcine immunoglobulin purification

Qilei Zhang, Tiantian Zhuang, Hongfei Tong, Hongyin Wang, Dongqiang Lin, Shanjing Yao

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China

Received:2014-10-13 Revised:2015-05-06 Online:2016-01-28 Published:2016-02-23
Contact: Dongqiang Lin
Supported by:
Supported by the National Natural Science Foundation of China (21276228 and 21476198), the Natural Science Foundation of Zhejiang Province (LR12B06003), and the Fundamental Research Funds for the Central Universities (2013QNA4032).

摘要/Abstract

摘要： Three hydrophobic charge-induction adsorbents with functional ligands of 4-mercapto-ethyl-pyridine, 2-mercapto-methyl-imidazole or 2-mercapto-benzimidazole were evaluated in the purification of porcine immunoglobulin from porcine blood. Adsorption isothermswere studied under different pH conditions. The adsorbent with 2-mercapto-methyl-imidazole as the ligand showed reasonable adsorption capacity (43.60 mg·g^-1 gel) with great selectivity and it also showed the best elution performance in chromatographic studies. A multi-pH step elution process was proposed for the 2-mercapto-methyl-imidazole adsorbent, and the results showed that high immunoglobulin purity (94.3%) and a yield of 9.8 mg·(ml plasma)^-1 could be achieved under the optimal condition of loading (pH 5.0)-pre-elution (pH 7.0)-elution (pH 3.8). Moreover, molecular simulation was employed to help in analyzing the binding mechanism between the ligands and immunoglobulin, and the results showed that both 2-mercapto-benzimidazole and 2-mercapto-methyl-imidazole ligands were docked on the same pocket (around TYR319 and LEU309) of the Fc fragment of immunoglobulin, with 2-mercaptobenzimidazole showing stronger binding interactions.

关键词: Hydrophobic charge-induction chromatography, Porcine immunoglobulin, Step elution, Molecular simulation

Abstract: Three hydrophobic charge-induction adsorbents with functional ligands of 4-mercapto-ethyl-pyridine, 2-mercapto-methyl-imidazole or 2-mercapto-benzimidazole were evaluated in the purification of porcine immunoglobulin from porcine blood. Adsorption isothermswere studied under different pH conditions. The adsorbent with 2-mercapto-methyl-imidazole as the ligand showed reasonable adsorption capacity (43.60 mg·g^-1 gel) with great selectivity and it also showed the best elution performance in chromatographic studies. A multi-pH step elution process was proposed for the 2-mercapto-methyl-imidazole adsorbent, and the results showed that high immunoglobulin purity (94.3%) and a yield of 9.8 mg·(ml plasma)^-1 could be achieved under the optimal condition of loading (pH 5.0)-pre-elution (pH 7.0)-elution (pH 3.8). Moreover, molecular simulation was employed to help in analyzing the binding mechanism between the ligands and immunoglobulin, and the results showed that both 2-mercapto-benzimidazole and 2-mercapto-methyl-imidazole ligands were docked on the same pocket (around TYR319 and LEU309) of the Fc fragment of immunoglobulin, with 2-mercaptobenzimidazole showing stronger binding interactions.

Key words: Hydrophobic charge-induction chromatography, Porcine immunoglobulin, Step elution, Molecular simulation

Qilei Zhang, Tiantian Zhuang, Hongfei Tong, Hongyin Wang, Dongqiang Lin, Shanjing Yao. Experimental and in silico studies on three hydrophobic charge-induction adsorbents for porcine immunoglobulin purification[J]. Chinese Journal of Chemical Engineering, 2016, 24(1): 151-157.

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Experimental and in silico studies on three hydrophobic charge-induction adsorbents for porcine immunoglobulin purification

Experimental and in silico studies on three hydrophobic charge-induction adsorbents for porcine immunoglobulin purification

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