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题名: High-performance affinity chromatography with immobilization of protein A and L-histidine on molded monolith
作者: Luo, QZ;  Zou, HF;  Zhang, Q;  Xiao, XZ;  Ni, JY
关键词: affinity chromatography ;  monolith ;  protein A ;  L-histidine ;  human IgG
刊名: BIOTECHNOLOGY AND BIOENGINEERING
发表日期: 2002-12-05
DOI: 10.1002/bit.10391
卷: 80, 期:5, 页:481-489
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology
研究领域[WOS]: Biotechnology & Applied Microbiology
英文摘要: Reactive monoliths of macroporous poly(glycidyl methacrylate-co-ethylene dimethacrylate) have been prepared by "in-situ" copolymerization of the monomers in the presence of porogenic diluents. Protein A and L-histidine were immobilized on the monoliths directly or through a spacer arm, respectively. The properties of these two kinds of affinity columns were characterized, and the results showed that the columns with coupling of ligands by a spacer arm have some extent of non-specific adsorption for bovine serum albumin. The affinity column based on the monolithic polymer support provided us with good hydrodynamic characteristic, low flow resistance, and easy preparation. These two affinity columns were used for the purification of immunoglobulin G from human serum. The purity of the purified IgG was detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The stability of the protein A affinity column was investigated, and its performance remained invariable after half a year. The effects of the nature and the pH of the buffer system on the adsorption capacity of human IgG on histidyl affinity column were also investigated. The protein A affinity column is favorable for rapid analysis of human IgG samples. In contrast, the advantages of mild elution conditions, high stability, as well as low cost provide the histidyl column further potential possibility for fast removal of IgG from human plasma in clinical applications. (C) 2002 Wiley Periodicals, Inc.
关键词[WOS]: HOLLOW-FIBER MEMBRANES ;  REVERSED-PHASE CHROMATOGRAPHY ;  CO-ETHYLENE DIMETHACRYLATE) ;  IMMUNOGLOBULIN-G ;  EXCHANGE CHROMATOGRAPHY ;  PSEUDOBIOAFFINITY CHROMATOGRAPHY ;  ADSORPTION CAPACITY ;  STATIONARY-PHASE ;  SEPARATION ;  PURIFICATION
语种: 英语
WOS记录号: WOS:000178822100001
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/139081
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.Acad Sinica, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116011, Peoples R China

Recommended Citation:
Luo, QZ,Zou, HF,Zhang, Q,et al. High-performance affinity chromatography with immobilization of protein A and L-histidine on molded monolith[J]. BIOTECHNOLOGY AND BIOENGINEERING,2002,80(5):481-489.
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