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A novel protein refolding method integrating ion exchange chromatography with artificial molecular chaperone 被引量:3
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作者 Qin Ming Zhang Chao Zhan Wang Jiang Feng Liu Li Li Wang 《Chinese Chemical Letters》 SCIE CAS CSCD 2008年第5期595-598,共4页
Artificial molecular chaperone (AMC) and ion exchange chromatography (IEC) were integrated, thus a new refolding method, artificial molecular chaperone-ion exchange chromatography (AMC-IEC) was developed. Compar... Artificial molecular chaperone (AMC) and ion exchange chromatography (IEC) were integrated, thus a new refolding method, artificial molecular chaperone-ion exchange chromatography (AMC-IEC) was developed. Compared with AMC and IEC, the activity recovery of lysozyme obtained by AMC-IEC was much higher in the investigated range of initial protein concentrations, and the results show that AMC-IEC is very efficient for protein refolding at high concentrations. When the initial concentration of lysozyme is 180 mg/mL, its activity recovery obtained by AMC-IEC is still as high as 76.6%, while the activity recoveries obtained by AMC and IEC are 45.6% and 42.4%, respectively. 展开更多
关键词 Artificial molecular chaperone Ion exchange chromatography protein refolding LYSOZYME protein folding liquid chromatography
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重组人干扰素-γ的制备与鉴定 被引量:3
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作者 吴丹 高栋 +1 位作者 白泉 耿信笃 《色谱》 CAS CSCD 北大核心 2008年第2期206-211,共6页
用聚乙二醇200疏水相互作用色谱固定相(PEG200-STHIC)分别在色谱柱和色谱饼上完成了一步复性并同时纯化来源于大肠杆菌(E.coli)表达的重组人干扰素-γ(rhIFN-γ)。为了能使色谱分离方法用于不同来源的rhIFN-γ的纯化,对rhIFN-γ在反相... 用聚乙二醇200疏水相互作用色谱固定相(PEG200-STHIC)分别在色谱柱和色谱饼上完成了一步复性并同时纯化来源于大肠杆菌(E.coli)表达的重组人干扰素-γ(rhIFN-γ)。为了能使色谱分离方法用于不同来源的rhIFN-γ的纯化,对rhIFN-γ在反相色谱、离子交换色谱、固定化镍离子亲和色谱上的保留行为也进行了研究。色谱柱纯化的rhIFN-γ收集液经排阻色谱除盐和冷冻干燥得到rhIFN-γ干粉。用基质辅助激光解吸电离飞行时间质谱对rhIFN-γ干粉进行了测定,rhIFN-γ单体的相对分子质量为17184.0,二聚体的相对分子质量为34204.4。用细胞病变抑制法(CPEI)测定rhIFN-γ干粉的比活性为9.5×108IU/mg。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)测定rhIFN-γ干粉的纯度高于95%。用色谱柱复性并同时纯化rhIFN-γ的质量回收率达到93.7%,纯度高于95%,比活性为4.3×107IU/mg。结果表明,采用PEG200-STHIC色谱柱复性并同时纯化rhIFN-γ是一种十分高效的方法。 展开更多
关键词 重组人干扰素-γ 蛋白折叠液相色谱法 高效疏水色谱 复性 纯化
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源于大肠杆菌蛋白的表达、液相色谱复性与纯化新进展 被引量:9
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作者 王骊丽 耿信笃 《中国科学(B辑)》 CSCD 北大核心 2009年第8期711-727,共17页
对近两年来源于大肠杆菌(Escherichia coli,E.coli)的蛋白表达和用蛋白折叠液相色谱(protein folding liquid chrom atography,PFLC)法对所形成的包涵体目标蛋白的复性并同时纯化的新近发展做了简要的介绍和评述.PFLC法用于包涵体蛋白... 对近两年来源于大肠杆菌(Escherichia coli,E.coli)的蛋白表达和用蛋白折叠液相色谱(protein folding liquid chrom atography,PFLC)法对所形成的包涵体目标蛋白的复性并同时纯化的新近发展做了简要的介绍和评述.PFLC法用于包涵体蛋白分离、纯化很广,其特点是除了在色谱柱上将目标蛋白与其他组分分开,还同时要在色谱柱上进行包涵体蛋白折叠.可以说,现代生物技术中所用的大多数有价值蛋白产品的制备仍然有赖于不同机理的液相色谱(LC)法.而用PFLC法对源于E.coli的蛋白的制备方法更具可塑性和容易达到规模化,其生成本可以成倍地降低.该文主要内容包括了E.coli蛋白的表达及样品前处理、PFLC的实用范围、PFLC的优化、PFLC中的新技术、新设备和新方法、PFLC的分子学机理、应用事例及对未来的展望. 展开更多
关键词 大肠杆菌 蛋白折叠液相色谱法 蛋白复性 蛋白纯化 液相色谱 生物工程
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Recent developments of the stoichiometric displacement model for separation processes 被引量:2
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作者 GENG Xindu Fred E.Regnier WANG Yan 《Chinese Science Bulletin》 SCIE EI CAS 2001年第21期1763-1772,共10页
Substantial attention has been paid to the stoi-chiometric displacement models for solute retention (SDM-R) in liquid chromatography and adsorption (SDM-A) at surfaces since they were developed. The SDM has a strongly... Substantial attention has been paid to the stoi-chiometric displacement models for solute retention (SDM-R) in liquid chromatography and adsorption (SDM-A) at surfaces since they were developed. The SDM has a strongly theoretical basis and has evolved to the point that it is widely applied in chemistry, biochemistry, molecular biology and biotechnology. This review introduces the history, recent developments, and new concepts relating to the SDM, including theory, mathematical expressions, and applications across a broad range of fields. 展开更多
关键词 liquid chromatography liquid-solid system stoichiomet- ric DISPLACEMENT RETENTION model adsorption mechanism protein folding.
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