کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1203001 1493574 2014 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Application of cyclodextrin-based eluents in hydrophobic charge-induction chromatography: Elution of antibody at neutral pH
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Application of cyclodextrin-based eluents in hydrophobic charge-induction chromatography: Elution of antibody at neutral pH
چکیده انگلیسی


• Competitive elution for HCIC is based on MEP/β-CD supermolecular interaction.
• MEP/β-CD inclusion complex relies on hydrophobic interaction and H-bond.
• 15 mM β-CD allows IgG elution from HCIC at neutral pH.

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 HCIC, 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 β-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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1352, 25 July 2014, Pages 62–68
نویسندگان
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