کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1199672 1493557 2014 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Salt-independent hydrophobic displacement chromatography for antibody purification using cyclodextrin as supermolecular displacer
ترجمه فارسی عنوان
کروماتوگرافی جابجایی هیدروفی فیزیکی مستقل از نمک برای تصفیه آنتی بادی با استفاده از سیکلوکودکسترین به عنوان غده فوق کلیوی
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• Hydrophobicity-intensified HIC resins can allow the capture of antibody at the physiological salt concentration.
• Displacement elution of HIC can be achieved through phenyl/β-CD supermolecular interaction.
• β-CD has an association constant (Ka) of 4.1 × 103 M−1 with the ligand.
• Salt-independent hydrophobic displacement chromatography has proven effective in direct purification of IgG from human serum.

Hydrophobic interaction chromatography (HIC) offers an orthogonal selectivity to ion exchange chromatography and the combination of the two processes can provide a potential cost-effective alternative to protein A chromatography in industrial antibody purification. However, the application of HIC is limited by its close dependence on high concentrations of kosmotropic salts to achieve desired separation. These salts can cause antibody precipitation and induce the corrosion of manufacturing facilities. Here, we report a new strategy of salt-independent HIC, which can capture antibody at the physiological salt concentration and allow the recovery of bound proteins through cyclodextrin (CD)-based displacement elution. Hydrophobicity-intensified HIC media with different coupling amount of phenyl ligands were prepared and assessed for their antibody binding capacity and selectivity. β-CD was investigated for its supermolecular interaction with phenyl ligands and elution capacity as a displacer. The results clarified a nearly linear correlation between binding capacity of human immunoglobulin G (IgG) and phenyl coupling density in the range of 44–159 μmol/mL. The host–guest interaction between β-CD and the phenyl ligands revealed a modest binding strength (Ka = 4.1 × 103 M−1), and 15 mM β-CD solution showed a general effectiveness as displacement eluent for these HIC media, with IgG recovery varying with the ligand density. This strategy allowed the direct purification of human IgG from serum with satisfactory purity. The whole procedure of this method, including loading and elution, can be performed under physiological conditions. We expect such a salt-independent mode of HIC could be used as a capture or intermediate step in industrial antibody purification.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1369, 21 November 2014, Pages 98–104
نویسندگان
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