Separation of proteins from human plasma by sample displacement chromatography in hydrophobic interaction mode.
Separation of proteins from human plasma by sample displacement chromatography in hydrophobic interaction mode.
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样品置换色谱法以疏水相互作用模式将蛋白质与人血浆分离。
DOI：
10.1002/elps.201200006
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发表时间：
2012-07
期刊：
影响因子：
2.9
通讯作者：
Mueller, Egbert
中科院分区：
文献类型：
--
作者：
Josic, Djuro;Breen, Lucas;Clifton, James;Gajdosik, Martina Srajer;Gaso-Sokac, Dajana;Rucevic, Marijana;Mueller, Egbert
反相和离子交换模式的样品置换色谱（SDC）大约在20年前被引入。该方法首次用于多肽和蛋白质的制备纯化。最近，离子交换模式下的SDC也成功地用于从人血浆中富集低丰度蛋白质。本文演示了在疏水相互作用模式下使用SDC分离血浆蛋白。通过在样品应用过程中使用串联耦合的两个或多个柱，随后平行洗脱分离的柱，实现了结合蛋白的额外分离。进一步的低丰度、生理活性蛋白可以通过ESI-MS/MS进行高富集和检测。
Sample displacement chromatography (SDC) in reversed-phase and ion-exchange modes was introduced approximately twenty years ago. This method was first used for the preparative purification of peptides and proteins. Recently, SDC in ion-exchange mode was also successfully used for enrichment of low abundance proteins from human plasma. In this paper, the use of SDC for the separation of plasma proteins in hydrophobic interaction mode is demonstrated. By use of two or more columns coupled in series during sample application, and subsequent elution of detached columns in parallel, additional separation of bound proteins was achieved. Further low-abundance, physiologically active proteins could be highly enriched and detected by ESI-MS/MS.
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