SEIBUTSU BUTSURI KAGAKU

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Recent progress in mass spectrometry has made a major impact for discovery of plasma biomarkers. Although sensitivity of mass spectrometer for proteomics seemed to be very high, it still remained to be limited in quantification. This has hampered to validate molecular candidates in clinical samples. A validation system with high efficiency and throughput has been expected.
Multiple reaction monitoring (MRM) is a unique mode for quantitative analysis in mass spectrometry and being useful to quantify known analytes. We employed MRM as a mean of validation on possible marker peptides obtained by Q-TOF screening with non-labeled plasma proteins, and found that MRM validation in combination with Q-TOF screening might be an useful method to explore plasma biomarkers. Plasma proteins of mouse model for postmenopausal osteoporosis were separated by SDS-PAGE and in-gel digested to peptide with trypsin. These peptides were compared to those of control mice by ESI-Q-TOF. Mass spectra more than 3 fold increase in disease model were assigned by MS / MS analysis and used to construct MRM channels. The resulting MRM channels were directly used to validate their discriminatory power to distinguish disease conditions from control.
Construction of MRM channels with spectra from peptide assignment and direct use of MRM channels in biomarker validation appeared to be very effective. Collectively, MRM validation in combination with Q-TOF screening might be a suitable setting for a discovery of biomarker in any kind of body fluid including plasma.

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