HST-MRM-MS: A Novel High-Sample-Throughput Multiple Reaction Monitoring Mass Spectrometric Method for Multiplex Absolute Quantitation of Hepatocell…
Authors of this article are:
Jiang H, Zhang L, Zhang Y, Xie L, Wang Y, Lu H.
A summary of the article is shown below:
Absolute quantification of clinical biomarkers by mass spectrometry (MS) has been challenged due to low sample-throughput of current multiple reaction monitoring (MRM) methods. To overcome this problem, 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-termini and C-termini of the Lys C digested peptides respectively. The triplex dimethyl labeling produces three co-elute peaks in MRM traces, while the iTRAQ labeling produces 8 peaks in MS2, resulting 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). Besides, 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.
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