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题名: Protein A immobilized monolithic capillary column for affinity chromatography
作者: Pan, ZF;  Zou, HF;  Mo, WM;  Huang, XD;  Wu, RN
关键词: affinity chromatography ;  monolithic rod ;  capillary column ;  protein A ;  hIgG
刊名: ANALYTICA CHIMICA ACTA
发表日期: 2002-08-21
卷: 466, 期:1, 页:141-150
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: Monolithic capillary columns for affinity chromatography were prepared by an in situ polymerization procedure using glycidyl methacrylate (GMA) as a monomer and trimethylolpropane trimethacrylate (TRIM) and ethylene dimethacrylate (EDMA) as cross-linkers, respectively. Scanning electron microscopy was applied to characterize the morphology of the end of monolithic capillary and mercury intrusion porosimetry to characterize the polymer rod prepared within the confines of a stainless steel column with 50 mm x 4.6 mm i.d. under the same polymerization condition. Obvious differences in the porous properties between the TRIM- and EDMA-based monoliths could be observed. Moreover, the mechanical stability of these two monolithic capillary columns was compared by testing the reproducibility of the column performance. The rod prepared with GMA and TRIM proved to be mechanically more stable than that prepared with GMA and EDMA. Protein A was immobilized on the monolithic rod for affinity chromatography and the experiments were performed on a capillary electrophoresis instrument, using its pressure system as the driving force. Non-specific adsorption was not observed on the TRIM-based affinity column, as proved with bovine serum albumin (BSA) as a test protein. The affinity column prepared with GMA and TRIM was then applied to determine the hIgG concentration in human serum. The correlative coefficient of the calibration curve reached 0.9942. The amount of adsorbed hIgG was unaffected by the flow rate of the loading buffer, which makes this method suitable for fast determination of biomacromolecules in microliter samples. (C) 2002 Elsevier Science B.V All rights reserved.
关键词[WOS]: PERFORMANCE LIQUID-CHROMATOGRAPHY ;  RIGID POLYMER MONOLITHS ;  STATIONARY PHASES ;  SEPARATION MEDIA ;  POROUS PROPERTIES ;  CONTINUOUS BEDS ;  EXCHANGE CHROMATOGRAPHY ;  MACROPOROUS POLYMERS ;  ELECTROCHROMATOGRAPHY ;  CHEMISTRY
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000177682900017
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/84217
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116011, Peoples R China
2.Zhejiang Univ Technol, Coll Chem Engn, Zheijang 310014, Peoples R China

Recommended Citation:
Pan, ZF,Zou, HF,Mo, WM,et al. Protein A immobilized monolithic capillary column for affinity chromatography[J]. ANALYTICA CHIMICA ACTA,2002,466(1):141-150.
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