Simple and Sensitive Analysis for Dried Blood Spot Proteins by Sodium Carbonate Precipitation for Clinical Proteomics.,Journal of Proteome Research

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Simple and Sensitive Analysis for Dried Blood Spot Proteins by Sodium Carbonate Precipitation for Clinical Proteomics.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-04-28 , DOI: 10.1021/acs.jproteome.0c00271
Daisuke Nakajima ₁ , Yusuke Kawashima ₁ , Hirofumi Shibata ₂ , Takahiro Yasumi ₂ , Masahiko Isa ₂ , Kazushi Izawa ₂ , Ryuta Nishikomori ₃ , Toshio Heike _{2,

4} , Osamu Ohara ₁

Affiliation

Dried blood spots (DBS) are widely used for screening biomolecular profiles, including enzymatic activities. However, detection of minor proteins in DBS by liquid chromatography–mass spectrometry (LC-MS/MS) without pre-enrichment remains challenging because of the coexistence of large quantities of hydrophilic proteins. In this study, we address this problem by developing a simple method using sodium carbonate precipitation (SCP). SCP enriches hydrophobic proteins from DBS, allowing substantial removal of soluble proteins. In combination with SCP, we used quantitative LC-MS/MS proteome analysis in a data-independent acquisition mode (DIA) to enhance the sensitivity and quantification limits of proteome analysis. As a result, identification of 1977 proteins in DBS is possible, including 585 disease-related proteins listed in the Online Mendelian Inheritance in Man.

中文翻译：

用于蛋白质组学的碳酸钠沉淀法对干血斑蛋白的简单而灵敏的分析。

干血斑（DBS）被广泛用于筛选生物分子特征，包括酶活性。然而，由于大量亲水性蛋白质的共存，通过液相色谱-质谱法（LC-MS / MS）检测DBS中的微量蛋白质仍然具有挑战性。在这项研究中，我们通过开发一种使用碳酸钠沉淀（SCP）的简单方法解决了这个问题。SCP从DBS富集疏水蛋白，从而可大量去除可溶性蛋白。与SCP结合使用时，我们在数据独立获取模式（DIA）中使用了定量LC-MS / MS蛋白质组分析，以增强蛋白质组分析的灵敏度和定量限。结果，可以鉴定DBS中的1977种蛋白质，

更新日期：2020-07-02

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