Selectivity of monolithic supports under overloading conditions and their use for separation of human plasma and isolation of low abundance proteins
Selectivity of monolithic supports under overloading conditions and their use for separation of human plasma and isolation of low abundance proteins
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DOI：
10.1016/j.chroma.2010.11.059
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发表时间：
2011-04-29
影响因子：
4.1
通讯作者：
Josic, Dj.
中科院分区：
文献类型：
--
作者：
Brgles, Marija;Clifton, James;Josic, Dj.
人血清白蛋白（HSA）和免疫球蛋白G（IgG）占人血浆中所有蛋白质的75%以上。这两种蛋白质经常干扰低丰度蛋白质的检测、测定和纯化，所述低丰度蛋白质可以是各种疾病的潜在生物标志物和生物标志物候选物。一些低丰度血浆蛋白如凝血因子和抑制剂也是重要的治疗剂。本文采用样品置换色谱法（SDC）对离子交换整体载体在超负荷条件下的性能进行了表征。如果这些支持物用于分离人血浆，则在阴离子和阳离子交换模式下结合和洗脱的蛋白质的组成取决于柱负载。在超负荷条件下，弱结合的蛋白质，如阴离子交换中的HSA和阳离子交换模式中的IgG被较强结合的蛋白质取代，并且这种现象不依赖于柱的大小。因此，具有100和200 μ L的柱体积的小型整体柱是高通量筛选的理想支持物，以开发用于分离复杂混合物的新方法，以及用于蛋白质组学技术中的样品制备。由爱思唯尔公司出版
Human serum albumin (HSA) and immunoglobulin G (IgG) represent over 75% of all proteins present in human plasma. These two proteins frequently interfere with detection, determination and purification of low abundance proteins that can be potential biomarkers and biomarker candidates for various diseases. Some low abundance plasma proteins such as clotting factors and inhibitors are also important therapeutic agents. In this paper, the characterization of ion-exchange monolithic supports under overloading conditions was performed by use of sample displacement chromatography (SDC). If these supports were used for separation of human plasma, the composition of bound and eluted proteins in both anion- and cation-exchange mode is dependent on column loading. Under overloading conditions, the weakly bound proteins such as HSA in anion-exchange and IgG in cation-exchange mode are displaced by stronger binding proteins, and this phenomenon was not dependent on column size. Consequently, small monolithic columns with a column volume of 100 and 200 mu L are ideal supports for high-throughput screening in order to develop new methods for separation of complex mixtures, and for sample preparation in proteomic technology. Published by Elsevier B.V.