S-Trap, an Ultrafast Sample-Preparation Approach for Shotgun Proteomics,Journal of Proteome Research

当前位置： X-MOL 学术 › J. Proteome Res. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

S-Trap, an Ultrafast Sample-Preparation Approach for Shotgun Proteomics
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-08-28 , DOI: 10.1021/acs.jproteome.8b00505
Milkessa HaileMariam _{1,

2} , Rodrigo Vargas Eguez ₁ , Harinder Singh ₁ , Shiferaw Bekele ₁ , Gobena Ameni ₂ , Rembert Pieper ₁ , Yanbao Yu ₁

Affiliation

The success of shotgun proteomic analysis depends largely on how samples are prepared. Current approaches (such as those that are gel-, solution-, or filter-based), although being extensively employed in the field, are time-consuming and less effective with respect to the repetitive sample processing, recovery, and overall yield. As an alternative, the suspension trapping (S-Trap) filter has been commercially available very recently in the format of a single or 96-well filter plate. In contrast to the conventional filter-aided sample preparation (FASP) approach, which utilizes a molecular weight cut-off (MWCO) membrane as the filter and requires hours of processing before digestion-ready proteins can be obtained, the S-Trap employs a three-dimensional porous material as filter media and traps particulate protein suspensions with the subsequent depletion of interfering substances and in-filter digestion. Due to the large (submicron) pore size, each centrifugation cycle of the S-Trap filter only takes 1 min, which significantly reduces the total processing time from approximately 3 h by FASP to less than 15 min, suggesting an ultrafast sample-preparation approach for shotgun proteomics. Here, we comprehensively evaluate the performance of the individual S-Trap filter and 96-well filter plate in the context of global protein identification and quantitation using whole-cell lysate and clinically relevant sputum samples.

中文翻译：

S-Trap，一种用于Shot弹枪蛋白质组学的超快速样品制备方法

shot弹枪蛋白质组学分析的成功在很大程度上取决于样品的制备方式。当前的方法（例如基于凝胶，溶液或过滤器的方法）虽然在本领域得到了广泛采用，但在重复性样品处理，回收和总产率方面却很耗时且效率较低。作为替代方案，悬浮液捕获（S-Trap）过滤器最近已经以单孔或96孔滤板的形式在市场上出售。与常规的过滤器辅助样品制备（FASP）方法不同，该方法利用分子量截留（MWCO）膜作为过滤器，并且需要数小时的处理才能获得可消化的蛋白质，S-Trap采用三维多孔材料作为过滤介质，捕获颗粒状蛋白质悬浮液，随后消耗掉干扰物质并进行过滤器内消化。由于孔径大（亚微米），S-Trap过滤器的每个离心周期仅需1分钟，这将总处理时间从FASP的大约3小时显着减少到不到15分钟，这表明样品制备方法超快用于shot弹枪蛋白质组学。在这里，我们在使用全细胞裂解物和临床相关痰液样本进行全球蛋白质鉴定和定量分析的背景下，全面评估了单个S-Trap过滤器和96孔滤板的性能。S-Trap过滤器的每个离心周期仅花费1分钟，这将总处理时间从FASP的大约3小时减少到不到15分钟，这表明，弹枪蛋白质组学的超快速样品制备方法。在这里，我们在使用全细胞裂解物和临床相关痰液样本进行全球蛋白质鉴定和定量分析的背景下，全面评估了单个S-Trap过滤器和96孔滤板的性能。S-Trap过滤器的每个离心周期仅花费1分钟，这将总处理时间从FASP的大约3小时减少到不到15分钟，这表明，弹枪蛋白质组学的超快速样品制备方法。在这里，我们在使用全细胞裂解物和临床相关痰液样本进行全球蛋白质鉴定和定量分析的背景下，全面评估了单个S-Trap过滤器和96孔滤板的性能。

更新日期：2018-08-29

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11