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超大孔离子交换制备色谱分离纯化高活性凝血因子Ⅷ
Authors
余蓉
周月芳
+5 more
康丽梅
张焱
李由
罗坚
苏志国
Publication date
1 January 2012
Publisher
Abstract
建立了一条从人血浆中分离高活性凝血因子Ⅷ(FⅧ)的纯化工艺。基于FⅧ和介质孔径的尺度比及其对蛋白质活性影响的分析,设计了以超大孔离子交换制备色谱为核心步骤的新型分离纯化工艺。分别进行超大孔离子交换色谱与传统离子交换色谱的条件优化,并对优化工艺所得产品进行了活性检测(底物显色法)和纯度检测(高效凝胶过滤和凝胶电泳)。结果表明,超大孔介质结构不但可以有效地保护蛋白质大分子结构,而且能够大幅度地提高制备色谱的传质速率,从而得到具有高凝血活性的FⅧ产品。FⅧ在超大孔制备色谱过程中的回收率(85%)比传统离子交换制备色谱高4~5倍,产品比活高达154 IU/mg。此外,还研究了超大孔介质的再生程序,采用5个柱体积的1 mol/L NaOH低流速清洗色谱柱,保证了色谱工艺的稳定性。本纯化工艺步骤简单,重现性好,易于放大生产
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Institutional Repository of Institute of Process Engineering, CAS (IPE-IR)
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oai:ir.ipe.ac.cn:122111/46060
Last time updated on 11/04/2021
Institutional Repository of Institute of Process Engineering, CAS (IPE-IR)
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oai:ir.ipe.ac.cn:122111/32615
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Institutional Repository of Institute of Process Engineering, CAS (IPE-IR)
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oai:ir.ipe.ac.cn:122111/10012
Last time updated on 09/11/2016