当前位置:科学网首页 > 小柯机器人 >详情
科学家通过plexDIA提高灵敏蛋白质组学的通量
作者:小柯机器人 发布时间:2022/7/21 17:25:57

近日,美国东北大学Nikolai Slavov等研究人员合作通过plexDIA提高灵敏蛋白质组学的通量。2022年7月14日,国际知名学术期刊《自然—生物技术》在线发表了这一成果。

为了提高敏感蛋白质组学的通量,研究人员开发了一个实验和计算框架,称为plexDIA,用于同时对肽和样品进行多路分析。使用plexDIA的复用分析可以在不降低蛋白质组覆盖率或定量准确性的情况下,以倍数增加标签数量的通量。通过使用三复合非异构质量标签,plexDIA可以对纳克级样品中多三倍的蛋白质比率进行定量。使用1小时的活性梯度,plexDIA在每个标记的三联体样本中量化了约8,000个蛋白质,并提高了数据的完整性,将不同样本的缺失数据减少了2倍以上。

应用于单个人类细胞,plexDIA在每个细胞中量化了约1,000个蛋白质,并在每个细胞使用约5分钟的活性色谱时,在一个plexDIA组中实现了98%的数据完整性。这些结果建立了一个通用框架,可用于提高敏感和定量蛋白质分析的通量。

据了解,目前的质谱方法能够对大量的样品进行高通量的蛋白质组学分析,但低样品量的蛋白质组学分析在深度和通量上仍然受到限制。

附:英文原文

Title: Increasing the throughput of sensitive proteomics by plexDIA

Author: Derks, Jason, Leduc, Andrew, Wallmann, Georg, Huffman, R. Gray, Willetts, Matthew, Khan, Saad, Specht, Harrison, Ralser, Markus, Demichev, Vadim, Slavov, Nikolai

Issue&Volume: 2022-07-14

Abstract: Current mass spectrometry methods enable high-throughput proteomics of large sample amounts, but proteomics of low sample amounts remains limited in depth and throughput. To increase the throughput of sensitive proteomics, we developed an experimental and computational framework, called plexDIA, for simultaneously multiplexing the analysis of peptides and samples. Multiplexed analysis with plexDIA increases throughput multiplicatively with the number of labels without reducing proteome coverage or quantitative accuracy. By using three-plex non-isobaric mass tags, plexDIA enables quantification of threefold more protein ratios among nanogram-level samples. Using 1-hour active gradients, plexDIA quantified ~8,000 proteins in each sample of labeled three-plex sets and increased data completeness, reducing missing data more than twofold across samples. Applied to single human cells, plexDIA quantified ~1,000 proteins per cell and achieved 98% data completeness within a plexDIA set while using ~5minutes of active chromatography per cell. These results establish a general framework for increasing the throughput of sensitive and quantitative protein analysis.

DOI: 10.1038/s41587-022-01389-w

Source: https://www.nature.com/articles/s41587-022-01389-w

期刊信息

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:31.864
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex