Comparison of ultrafiltration units for proteomic and N-glycoproteomic analysis by the filter-aided sample preparation method
Comparison of ultrafiltration units for proteomic and N-glycoproteomic analysis by the filter-aided sample preparation method
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DOI：
10.1016/j.ab.2010.12.004
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发表时间：
2011-03-15
影响因子：
2.9
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Wisniewski, Jacek R.;Zielinska, Dorota F.;Mann, Matthias
过滤器辅助样品制备（FASP）方法允许对用去污剂溶解的生物样品进行无凝胶处理，以通过质谱法进行蛋白质组学分析。在FASP中，洗涤剂通过超滤去除，并且在蛋白质消化后，肽与未消化的材料分离。在这里，我们比较了不同的过滤设备的蛋白质组和糖蛋白质组分析的有效性。我们表明，Microcon和Vivacon过滤装置的标称分子量截留值为30，000和50，000（分别为30和50 k）同样适用于FASP，而Microcon 30 k装置最适合于N-糖基化位点的作图。使用具有这些相对大的截留值的过滤器有利于洗涤剂的消耗。(C)2010年爱思唯尔公司All rights reserved.
The filter-aided sample preparation (FASP) method allows gel-free processing of biological samples solubilized with detergents for proteomic analysis by mass spectrometry. In FASP detergents are removed by ultrafiltration, and after protein digestion peptides are separated from undigested material. Here we compare the effectiveness of different filtration devices for analysis of proteomes and glycoproteomes. We show that Microcon and Vivacon filtration units with nominal molecular weight cutoffs of 30,000 and 50,000 (30 and 50 k, respectively) are equally suitable for FASP, whereas Microcon 30 k units are most appropriate for mapping of N-glycosylation sites. The use of filters with these relatively large cutoffs facilitates depletion of detergents. (C) 2010 Elsevier Inc. All rights reserved.