Targeted Quantification of Peptides Using Miniature Mass Spectrometry
journal contributionposted on 06.04.2020, 09:13 by Spencer Chiang, Wenpeng Zhang, Charles Farnsworth, Yiying Zhu, Kimberly Lee, Zheng Ouyang
Proteomics by mass spectrometry (MS) allows for the identification of amino acid/peptide sequences in complex mixtures. Peptide analysis and quantitation enables screening of protein biomarkers and targeted protein biomarker analysis for clinical applications. Whereas miniature mass spectrometers have primarily demonstrated point-of-care analyses with simple procedures aiming at drugs and lipids, it would be interesting to explore their potential in analyzing proteins and peptides. In this work, we adapted a miniature MS instrument for peptide analysis. A mass range as wide as 100–2000 m/z was achieved for obtaining peptide spectra using this instrument with dual linear ion traps. MS2 and MS3 can be performed to analyze a wide range of peptides. The parameters of pressure, electric potentials, and solution conditions were optimized to analyze peptides with molecular weights between 900 and 1800 Da. The amino acid sequences were identified using both beam-type and in-trap collision-induced dissociation, and the results were comparable to those obtained by a commercial quadrupole time-of-flight mass spectrometer. With product ion monitoring scan mode, peptide quantitation was performed with a limit of detection of 20 nM achieved for the Met peptide. The method developed has also been applied to the analysis of the trypsin-digested cell lysate of SKBR3 cells with a low expression level of the Met gene.
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acid sequencesprotein biomarkersprotein biomarker analysisMet peptideMS instrumentpeptide analysismass spectrometrypeptide quantitationproduct ion monitoring scan modeMS 2MS 3Miniature Mass Spectrometry Proteomicspoint-of-care analyses20 nMSKBR 3 cellsmass rangein-trap collision-induced dissociationexpression leveltrypsin-digested cell lysateion trapssolution conditionsMet geneTargeted Quantificationmass spectrometers1800 DaPeptide analysispeptide spectraquadrupole time-of-flight mass spectrometer