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

详细信息    查看全文

• 作者：Qilei Zhang ; Tiantian Zhuang ; Hongfei Tong ; Hongyin Wang ; Dongqiang Lin ; ^{lindq@zju.edu.cn" class="auth_mail" title="E-mail the corresponding author} ; Shanjing Yao
• 关键词：Hydrophobic charge-induction chromatography ; Porcine immunoglobulin ; Step elution ; Molecular simulation
• 刊名：Chinese Journal of Chemical Engineering
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：24
• 期：1
• 页码：151-157
• 全文大小：923 K

文摘

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 isotherms were 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-mercapto-benzimidazole showing stronger binding interactions.