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Investigation and reduction of sub-microgram peptide loss using molecular weight cut-off fractionation prior to mass spectrometric analysis

机译:在质谱分析之前使用分子量截止分馏法研究和减少亚微克肽损失

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摘要

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 H _2O. The strategy presented here enhances recovery from MWCO separation for sub-μg peptide samples.
机译:这项工作研究了将甲醇和盐改性剂引入分子量截断膜式离心过滤器(MWCO),以丰富亚微克肽的量。使用神经肽标准缓激肽,无论是否存在未消化的蛋白质,样品损失都减少了两个数量级。另外,研究了牛血清白蛋白的胰蛋白酶消化。富集甲醇后,从MALDI质谱图中鉴定出27个胰蛋白酶肽,而使用H _2O进行MWCO分离后,仅鉴定了2个胰蛋白酶肽。此处提出的策略可提高MWCO分离中亚μg肽样品的回收率。

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