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Novel bifunctional material coupled to PASEF enables sensitive extracellular vesicles phosphoproteomic analysis of urine
Poster Title: Novel bifunctional material coupled to PASEF enables sensitive extracellular vesicles phosphoproteomic analysis of urine
Submitted on 05 Oct 2021
Author(s): Jie Sun1,2, W.Andy Tao3, Xiaoxian Du4 and Xianming Liu4
Affiliations: 1Southeast University, Nanjing, Jiangsu Province, China 2Bell Mountain Molecular Medicine, Nanjing, Jiangsu Province, China 3Purdue University, Indiana, United States 4Bruker Daltonics, Shanghai, China SouthMethods UniversitMethodsMethods NanjinMethods ProvinMethods ChinaMethods Methods Methods MediciMethods ProvinMethods ChinaMethods UniversityMethods Methods Methods Methods MethodsMethods Methods ChinaMethods
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Abstract: Here, we introduced a novel bifunctionalized magnetic beads (BiMBs) coupled trapped ion mobility (TIMS) and PASEF for phosphoproteomic analysis of urine samples (Fig. 1). Benefiting from the highly efficient beads together with highly sensitive timsTOF Pro, we can identify 36262 unique EV peptides corresponding to 3302 unique proteins and 3233 unique phosphopeptides representing 1098 unique phosphoproteins using only 100 μL and 5 mL urine samples, respectively. The EV proteome from BiMBs overlapped with 95 out of 100 top exosome marker proteins (Fig.2)
Furthermore, quantitative phosphoproteomics of urine samples from prostate cancer patients and healthy individuals revealed 121 upregulated phosphoproteins in cancer patients in contrast to the healthy group (Fig. 3). In addition, TIMS provides one more dimension of separation that enables to distinguish the isobaric peptides with different modified positions (Fig. 4).
Summary: •Highly efficient beads together with highly sensitive timsTOF Pro can achieve depth of proteome coverage even
with low amount urine sample
•Many upregulated phosphoproteins associate with prostate cancer according to quantitative phosphoproteomics analysis
•TIMS provides additional dimension of information on ions with specific CCS values, which may lead to broader applications with PTM analyses.
References: (1) Jie Sun, et. al., Synergistically Bifunctional Paramagnetic Separation Enables Efficient Isolation of Urine Extracellular Vesicles and Downstream Phosphoproteomic Analysis. ACS Applied Materials & Interfaces 2021, 13, 3, 3622-3630.Report abuse »
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