Investigation and reduction of sub-microgram peptide loss using molecular weight cut-off fractionation prior to mass spectrometric analysis
Investigation and reduction of sub-microgram peptide loss using molecular weight cut-off fractionation prior to mass spectrometric analysis
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DOI：
10.1002/jms.3069
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发表时间：
2012-10-01
影响因子：
2.3
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
--
作者：
Cunningham, Robert;Wang, Jingxin;Li, Lingjun
本研究将甲醇和盐改性剂引入分子量截止膜离心过滤器（MWCO）中，以富集亚微克肽量。使用神经肽标准，缓激肽，样品损失减少超过两个数量级，有无未消化的蛋白质存在。此外，还研究了牛血清白蛋白胰蛋白酶消化。甲醇富集后，MALDI质谱中鉴定出27个色氨酸肽，而水分离MWCO后，仅鉴定出2个色氨酸肽。本文提出的策略提高了亚肽MWCO分离样品的回收率。版权所有：John Wiley & Sons， Ltd。
This work investigates the introduction of methanol and a salt modifier to molecular weight cut-off membrane-based centrifugal filters (MWCO) to enrich sub-microgram peptide quantities. Using a neuropeptide standard, bradykinin, sample loss was reduced over two orders of magnitude with and without undigested protein present. Additionally, a bovine serum albumin tryptic digestion was investigated. Twenty-seven tryptic peptides were identified from MALDI mass spectra after enriching with methanol while only two tryptic peptides were identified after MWCO separation using H2O. The strategy presented here enhances recovery from MWCO separation for sub-mu g peptide samples. Copyright (c) 2012 John Wiley & Sons, Ltd.