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HST-MRM-MS: A Novel High-Sample-Throughput Multiple Reaction Monitoring Mass Spectrometric Method for Multiplex Absolute Quantitation of Hepatocellular Carcinoma Serum Biomarker
Journal of Proteome Research ( IF 4.4 ) Pub Date : 2018-10-31 , DOI: 10.1021/acs.jproteome.8b00790 Hucong Jiang , Lei Zhang , Ying Zhang , Liqi Xie , Yi Wang , Haojie Lu
Journal of Proteome Research ( IF 4.4 ) Pub Date : 2018-10-31 , DOI: 10.1021/acs.jproteome.8b00790 Hucong Jiang , Lei Zhang , Ying Zhang , Liqi Xie , Yi Wang , Haojie Lu
Absolute quantification of clinical biomarkers by mass spectrometry (MS) has been challenged due to low sample-throughput of current multiple reaction monitoring (MRM) methods. For this problem to be overcome, in this work, a novel high-sample-throughput multiple reaction monitoring mass spectrometric (HST-MRM-MS) quantification approach is developed to achieve simultaneous quantification of 24 samples. Briefly, triplex dimethyl reagents (L, M, and H) and eight-plex iTRAQ reagents were used to label the N- and C-termini of the Lys C-digested peptides, respectively. The triplex dimethyl labeling produces three coelute peaks in MRM traces, and the iTRAQ labeling produces eight peaks in MS2, resulting in 24 (3×8) channels in a single experiment. HST-MRM-MS has shown good accuracy (R2 > 0.98 for absolute quantification), reproducibility (RSD < 15%), and linearity (2–3 orders of magnitude). Moreover, the novel method has been successfully applied in quantifying serum biomarkers in hepatocellular carcinoma (HCC)-related serum samples. In conclusion, HST-MRM-MS is an accurate, high-sample-throughput, and broadly applicable MS-based absolute quantification method.
中文翻译:
HST-MRM-MS:一种用于肝癌血清生物标志物多重绝对定量的新型高通量多反应监测质谱方法
由于当前的多反应监测(MRM)方法的样品通量较低,因此通过质谱(MS)对临床生物标志物进行绝对定量的方法受到了挑战。为了克服这个问题,在这项工作中,开发了一种新颖的高样品通量多反应监测质谱(HST-MRM-MS)定量方法,以实现同时定量24个样品。简而言之,三重二甲基试剂(L,M和H)和八重iTRAQ试剂分别用于标记Lys C消化的肽的N和C末端。三联体二甲基标记在MRM迹线中产生三个共洗脱峰,而iTRAQ标记在MS2中产生八个峰,在单个实验中产生24(3×8)个通道。HST-MRM-MS表现出良好的准确性(R 2> 0.98(绝对定量),重现性(RSD <15%)和线性(2-3个数量级)。此外,该新方法已成功应用于定量肝细胞癌(HCC)相关血清样品中的血清生物标志物。总之,HST-MRM-MS是一种准确,高样品通量且广泛适用的基于MS的绝对定量方法。
更新日期:2018-11-02
中文翻译:
HST-MRM-MS:一种用于肝癌血清生物标志物多重绝对定量的新型高通量多反应监测质谱方法
由于当前的多反应监测(MRM)方法的样品通量较低,因此通过质谱(MS)对临床生物标志物进行绝对定量的方法受到了挑战。为了克服这个问题,在这项工作中,开发了一种新颖的高样品通量多反应监测质谱(HST-MRM-MS)定量方法,以实现同时定量24个样品。简而言之,三重二甲基试剂(L,M和H)和八重iTRAQ试剂分别用于标记Lys C消化的肽的N和C末端。三联体二甲基标记在MRM迹线中产生三个共洗脱峰,而iTRAQ标记在MS2中产生八个峰,在单个实验中产生24(3×8)个通道。HST-MRM-MS表现出良好的准确性(R 2> 0.98(绝对定量),重现性(RSD <15%)和线性(2-3个数量级)。此外,该新方法已成功应用于定量肝细胞癌(HCC)相关血清样品中的血清生物标志物。总之,HST-MRM-MS是一种准确,高样品通量且广泛适用的基于MS的绝对定量方法。