等压法
化学
试剂
等压标记
串联质量标签
多路复用
计算生物学
鉴定(生物学)
色谱法
定量蛋白质组学
串联质谱法
质谱法
蛋白质组学
计算机科学
生物化学
蛋白质质谱法
电信
物理
物理化学
生物
基因
热力学
植物
作者
Zhiyong Wu,Wei Xiang,Lin Huang,Shuwei Li,Xumin Zhang
标识
DOI:10.1021/acs.analchem.3c00237
摘要
Isobaric labeling has emerged as an indispensable quantitative proteomic approach for its unprecedented multiplexing capacity in a single analysis. Currently, different hyperplexing approaches have been developed to meet the demand for the increasing sample size in large-scale cohort analysis. In this report, we present a tribrid hyperplexing approach by the combinatorial use of three types of isobaric reagents, a novel isobaric tag 16-plex (IBT16) reagent and the widely used tandem mass tag (TMT; TMT11) and TMTpro (TMT18) reagents. After the determination of labeling efficiency and the optimization of testing conditions, we systematically evaluated the identification and quantification performance of the three labeling reagents in both independent and combinatorial manners using the mixtures of E. coli and HeLa peptides with different ratios. Our results reveal that the three reagents are quite similar in all testing aspects despite some differences, and the combination use of the three reagents could expand the multiplexing capacity to up to 45-plex. Furthermore, we conclude the advantages of IBT16 in the combination use and the preferred combinations for different practical applications. Data are available via ProteomeXchange with identifier PXD037498.
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