磷酸蛋白质组学
多路复用
化学
蛋白质组学
定量蛋白质组学
等压标记
计算生物学
吞吐量
生物信息学
计算机科学
生物化学
磷酸化
基因
蛋白激酶A
蛋白质磷酸化
生物
无线
电信
作者
Xiaofang Zhong,Qinying Yu,Fengfei Ma,Dustin C. Frost,Lei Lü,Zheng-Wei Chen,Henrik Zetterberg,Cynthia M. Carlsson,Ozioma C. Okonkwo,Lingjun Li
标识
DOI:10.1021/acs.analchem.8b04580
摘要
Absolute quantification in targeted proteomics is challenging due to a variety of factors, including low specificity in complex backgrounds, limited analytical throughput, and wide dynamic range. To address these problems, we developed a hybrid offset-triggered multiplex absolute quantification (HOTMAQ) strategy that combines cost-effective mass difference and isobaric tags to enable simultaneous construction of an internal standard curve in the MS1 precursor scan, real-time identification of peptides at the MS2 level, and mass offset-triggered accurate quantification of target proteins in synchronous precursor selection (SPS)-MS3 spectra. This approach increases the analytical throughput of targeted quantitative proteomics by up to 12-fold. The HOTMAQ strategy was employed to verify candidate protein biomarkers in preclinical Alzheimer's disease with high accuracy. The greatly enhanced throughput and quantitative performance, paired with sample flexibility, makes HOTMAQ broadly applicable to targeted peptidomics, proteomics, and phosphoproteomics.
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