High-sensitivity profiling of glycoproteins from human blood serum through multiple-lectin affinity chromatography and liquid chromatography/tandem mass spectrometry
High-sensitivity profiling of glycoproteins from human blood serum through multiple-lectin affinity chromatography and liquid chromatography/tandem mass spectrometry
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DOI：
10.1016/j.jchromb.2006.07.067
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发表时间：
2007-01-01
影响因子：
3
通讯作者：
Novotny, Milos V.
中科院分区：
文献类型：
--
作者：
Madera, Milan;Mechref, Yehia;Novotny, Milos V.
我们在这里报告了使用一系列二氧化硅结合的凝集素在微尺度水平上使用高性能凝集素亲和富集糖蛋白。这种方法在从微升体积的人血清中富集糖蛋白方面的潜力已得到证实。将样品单独注射到填充有四种具有不同聚糖特异性的凝集素材料（Con A、SNA-I、UEA-I、PHA-L）的亲和微柱中，然后进行离线反相预分级和 Nano-LC/MS/MS，可以鉴定凝集素结合级分中的 108 种蛋白质，浓度动态范围为 7-10 个数量级。相比之下，多凝集素微柱亲和层析（另一种富集方法）只能鉴定 67 种蛋白质。微量水平的高性能凝集素亲和色谱的一个吸引人的特点是显着减少了样品损失，而这种损失通常是在较大样品量所需的大量样品制备过程中遇到的。 (c) 2006 Elsevier B.V. 保留所有权利。
We report here the use of high-performance lectin affinity enrichment of glycoproteins at microscale levels using a series of silica-bound lectins. The potential of this approach is being demonstrated for the glycoprotein enrichment from microliter volumes of human blood serum. Individual injections of sample to the affinity microcolumns packed with four lectin materials with different glycan specificities (Con A, SNA-I, UEA-I, PHA-L), followed by off-line reversed-phase pre-fractionation and nano-LC/MS/MS, permitted identification of 108 proteins in the lectin-bound fractions spanning a concentration dynamic range of 7-10 orders of magnitude. In contrast, multi-lectin microcolumn affinity chromatography, an alternative enrichment approach allowed identification of only 67 proteins. An attractive feature of high-performance lectin affinity chromatography at microscale levels is the substantial reduction of sample losses that are commonly experienced with extensive sample preparation needed for larger sample volumes. (c) 2006 Elsevier B.V. All rights reserved.