A new purification process for goose immunoglobulin IgY(ΔFc) with hydrophobic charge-induction chromatography

被引:33
作者
Tong, Hong-Fei [1 ]
Lin, Dong-Qiang [1 ]
Pan, Yue [1 ]
Yao, Shan-Jing [1 ]
机构
[1] Zhejiang Univ, State Key Lab Chem Engn, Dept Chem & Biol Engn, Hangzhou 310027, Zhejiang, Peoples R China
基金
中国国家自然科学基金;
关键词
Hydrophobic charge-induction; chromatography; Immunoglobulin; Goose plasma; Purification; PROTEIN-A CHROMATOGRAPHY; DUCK IMMUNOGLOBULINS; ANTIBODY; LIGAND; BINDING; ADSORBENTS; ADSORPTION; CAPTURE; SERUM; IGY;
D O I
10.1016/j.bej.2011.06.010
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Hydrophobic charge-induction chromatography (HCIC) is a novel bioseparation technique, especially for antibody purification. In this study, HCIC was used for the purification of goose immunoglobulin IgY(Delta fc). IgY(Delta Fc) is a kind of unique immunoglobulin existing in waterfowl, which natively lacks C(H)3 and C(H)4 domains of heavy chains and has some particular therapeutic applications. A new HCIC ligand, 2-mercapto-1-methyl-imidazole (MMI), was coupled to the divinyl-sulfone-activated agarose matrix to prepare the HCIC gel. The adsorption isotherms of IgY(Delta Fc) were investigated at different pHs. The saturated capacity of IgY(Delta Fc) at pH 5.0 could reach 187.5 mg/ml gel, and the pH-dependent adsorption behaviors were found. After the goose plasma was pre-treated with caprylic acid to precipitate some impurities, the supernatant could be directly loaded onto the HCIC column for IgY(Delta Fc) separation. The operation conditions were optimized, including the loading pH, elution pH and the loading volume. High separation efficiency was obtained with final purity of 98.6% and the yield of 85.0%. The results indicate that the new process with HCIC can be a promising technology for the cost-effective separation of IgY(Delta Fc) from plasma. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码:205 / 211
页数:7
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