肾移植早期排异反应时尿液中肽段生物标志物的定量蛋白质组学分析方法及后续的SRM化验方法发展

　　简介：In this study, we describe the development of a workflow utilizing off-line size fractionation coupled with on-line liquid chromatography and high resolution tandem MS on an LTQ Orbitrap XL™ hybrid mass spectrometer specifically eared toward the identification of differentially expressed proteins and endogenous peptides in urine. Selected protein or peptide biomarkers identified in the high-resolution discovery workflow were subsequently quantified in targeted selective reaction monitoring (SRM) assays on a TSQ Quantum Ultra™ triple quadrupole mass spectrometer. The described quantitative workflows were used to analyze urine samples from normal, early rejection and acute humoral rejection transplant patients.

　　仪器：LTQ Orbitrap XL

　　结论：

　　• A robust workflow was created for the recovery of intact endogenous peptides (not generated by proteolytic cleavage ex vivo) from urine.

　　• Quantitative, label-free differential analysis was applied to high-resolution MS spectra to drive the selection of putative peptide markers for early transplant rejection.

　　• The high-resolution accurate mass MS/MS fragmentation data facilitated design of a targeted SRM assay, by yielding highly accurate fragment assignments on difficult to analyze endogenous peptides.

　　• A targeted SRM assay for one of the peptides in the panel was tested on the patient sample set and quantitative data were obtained.

　　• The described workflows can serve as