个人信息Personal Information
副教授
博士生导师
硕士生导师
性别:男
毕业院校:大连理工大学
学位:博士
所在单位:生物工程学院
学科:生物化工
办公地点:生物工程学院547室
电子邮箱:renjun@dlut.edu.cn
Application of cyclodextrin-based eluents in hydrophobic charge-induction chromatography: Elution of antibody at neutral pH
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论文类型:期刊论文
发表时间:2014-07-25
发表刊物:JOURNAL OF CHROMATOGRAPHY A
收录刊物:SCIE、EI、PubMed、Scopus
卷号:1352
页面范围:62-68
ISSN号:0021-9673
关键字:Hydrophobic charge-induction chromatography; Host-guest interaction; Cyclodextrin; Competitive elution; Immunoglobulin G
摘要:Hydrophobic charge-induction chromatography (HCIC) has emerged as a useful addition to Protein A chromatography for antibody purification due to its remarkable merits in cost and stability. However, the instability of antibody during acidic elution, which may cause inactivation and aggregation, is still a major concern for the efficiency of this method. The aim of this study is to develop a new strategy of competitive elution with inclusion complexes in HCIC, and to apply it to antibody elution under neutral pH conditions. Interactions between 4-mercaptoethylpyridine (MEP), a typical ligand of HOC, and four different types of cyclodextrins (CDs) were investigated by molecular docking; immunoglobulin G (IgG) elution capacities of CDs were characterized on MEP-based HCIC mediums. The results demonstrated the general effectiveness of CD-based eluents for HCIC. This type of displacement eluents could allow an efficient elution of bound antibody over a broad range of pH and ion strength. With 15 mM beta-CD, elution of human IgG was achieved at physiological pH, with an average IgG recovery of 87%. When this elution strategy was used to separate antibody directly from human serum, substantial elution of bound IgG could be obtained at pH 7.4, with product purity comparable to traditional method with an acidic buffer. We expect such method can be of special interest in developing HCIC elution strategy for the proteins like antibody that are sensitive to acidic conditions. (C) 2014 Elsevier B.V. All rights reserved.