A New Method For Proteome And Metabolome Pretreatment Of Dry Blood Samples

Proteins and metabolites in blood as the most important biomarkers have been broadly applied for clinical diagnosis.With the development of biomass spectrometry,proteomics and metabolomics analysis technology MS-based has been widely used in the large-scale analysis of biomarkers.Generally,proteomic and metabolomic analyses need to adopt different sample preparation steps,separation systems and mass spectrometry methods due to the differences in physico-chemical properties.Therefore,the omics analysis of a large number of clinical samples often has the problems of low throughput and difficult to control.Due to these limitation,the current clinical omics analysis is often based on the results of a single cohort for biological and disease phenotype analysis,which is prone to single conclusion and inaccurate.In recent years,multi-omics analysis combined with various big data types is a new strategy.This strategy helps to reveal the biological relationship in a deeper way,but also places higher demands on sample preparation and analysis techniques.In this study,we designed an integrated and simple sample preparation technology called Multi-omic-SISPROT.This technique treats dried plasma one-stop and simultaneously performs metabolomic and proteomic analyses.(1)Combining fingertip blood sampling,this method can easily enrich 1 ?L blood samples into centrifugal devices of dry blood in room temperature.(2)The method can integrate protein and metabolite for sample pretreatment in 3 h and analyze them using the same LC-MS/MS in 2 h,identifying more than 150 proteins and 160 metabolites.Using the same sample preparation device and analysis system can minimize the data deviation caused by different processes.In order to improve the detection ability of low abundance biomarkers,we further analyzed 10 reported cancer protein biomarkers using the targeted property spectrum detection mode MRM,and obtained a good signal-to-noise ratio and a maximum detection threshold of 5 pg/m L.(3)Moreover,the dried plasma samples enriched in the device could still reach a good identification level after being stored at room temperature for 24 h.Anyhow,Multi-omic-SISPROT technology will trace blood sample collection,transportation and integrated sample preparation and based on the same LC-MS system to multiple omics analysis together.The breadth and depth of plasma proteome and metabolome analysis are considered.This greatly improves subject participation and promotes the application of proteomics and metabolomics in precision medicine.