Pushing the Isotopic Envelope: When carrier channels pollute their neighbors' signals.
Pushing the Isotopic Envelope: When carrier channels pollute their neighbors' signals.
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推动同位素包络：当载波信道污染其邻居的信号时。
DOI：
10.1101/2024.04.15.587811
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Payne,SamuelH
中科院分区：
文献类型：
--
作者：
Peterson,Connor;Boekweg,Hannah;Presley,Eilenora;Payne,SamuelH
单个细胞是生物学的基本单位，了解它们的功能和相互作用对于提高我们对健康和疾病的理解至关重要。单细胞蛋白质组学已经引起了质谱学家的强烈兴趣，其目标是通过适应目前在大量样品中使用的技术来定量单细胞的蛋白质组学。迄今为止，大多数方法优化研究都致力于增加单细胞的蛋白质组覆盖率。一种突出的技术是使用等压标记将许多单个细胞多路复用到单个数据采集事件中。伴随着单个电池，一个标签通常用于许多电池的混合集，称为载波或升压通道。虽然这提高了多肽的识别率，但一些研究小组已经研究了随着更多的细胞包含在载体通道中，例如100倍或500倍，对定量准确性的影响。本文探讨了多路复用试剂中的杂质如何导致在错误通道中作为可测量信号观察到的不准确定量。我们发现载体和单细胞之间的严重丰度差异，加上试剂杂质，可以掩盖通常用于单细胞的几个通道。对于载流子数量的100倍及以上，这种污染可能与来自单个细胞的真实信号一样丰富。因此，我们建议将载波通道限制在最小数量，并平衡识别和量化的目标。
Individual cells are the foundational unit of biology, and understanding their functions and interactions is critical to advancing our understanding of health and disease. Single cell proteomics has seen intense interest from mass spectrometrists, with a goal of quantifying the proteome of single cells by adapting current techniques used in bulk samples. To date, most method optimizations research has worked towards increasing the proteome coverage of single cells. One prominent technique multiplexes many individual cells into a single data acquisition event using isobaric labels. Accompanying the single cells, one label is typically used for a mixed set of many cells, called a carrier or boost channel. Although this improves peptide identification rates, several groups have examined the impact on quantitative accuracy as more cells are included in the carrier channel, e.g. 100x or 500x. This manuscript explores how impurities in the multiplexing reagent can lead to inaccurate quantification observed as a measurable signal in the wrong channel. We discover that the severe abundance differential between carrier and single cell, combined with the reagent impurities, can overshadow several channels typically used for single cells. For carrier amounts 100x and above, this contamination can be as abundant as true signal from a single cell. Therefore, we suggest limiting the carrier channel to a minimal amount and balance the goals of identification and quantification.
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