3C-O2-1515 PDF
Development of a Targeted Quantitation System for Human Body Fluid Proteome using a Wheat Germ Cell-Free Synthesis System and Selected Reaction Monitoring Assay
Mass spectrometry-based protein quantitation of various human body fluids have shown immense potential for clinical biomarker discovery. In particular, targeted protein quantitation using selected reaction monitoring (SRM) shows much promise for the validation of biomarker proteins from body fluid samples of limited availability. However, this technique requires an initial time consuming development of assay parameters and reference datasets. In this study, we have developed a large-scale SRM assay targeting human body fluid proteins based on publicly available proteomics dataset. For the high-throughput production of a peptide reference library, we selected 2,201 targeted peptides representing 1,215 proteins from previously published studies, and in vitro synthesized them as peptide concatamers using a wheat germ cell-free system. Our study clearly demonstrated that the high-throughput production of reference peptide library using wheat germ cell-free system is an effective approach to establish a reference peptide library for developing an SRM assay that targets over 1,000 proteins experimentally.