Simple and Integrated Spintip-Based Technology Applied for Deep Proteome Profiling

Chen, Wendong1,3; Wang, Shuai4; Adhikari, Subash1; Deng, Zuhui4; Wang, Lingjue1; Chen, Lan1; Ke, Mi1; Yang, Pengyuan3; Tian, Ruijun1,2

2016-05-03

10.1021/acs.analchem.6b00631

ANALYTICAL CHEMISTRY   影响因子和分区

0003-2700

1520-6882

Great efforts have been taken for developing high-sensitive mass spectrometry (MS)-based proteomic technologies, among which sample preparation is one of the major focus. Here, a simple and integrated spintip-based proteomics technology (SISPROT) consisting of strong cation exchange beads and C-18 disk in one pipet tip was developed. Both proteomics sample preparation steps, including protein preconcentration, reduction, alkylation, and digestion, and reversed phase (RP)-based desalting and high-pH RP-based peptide fractionation can be achieved in a fully integrated manner for the first time. This easy-to-use technology achieved high sensitivity with negligible sample loss. Proteomic analysis of 2000 HEK 293 cells readily identified 1270 proteins within 1.4 h of MS time, while 7826 proteins were identified when 100000 cells were processed and analyzed within only 22 h of MS time. More importantly, the SISPROT can be easily multiplexed on a standard centrifuge with good reproducibility (Pearson correlation coefficient > 0.98) for both single-shot analysis and deep proteome profiling with five-step high-pH RP fractionation. The SISPROT was exemplified by the triplicate analysis of 100000 stern cells from human exfoliated deciduous teeth (SHED). This led to the identification of 9078 proteins containing 3771 annotated membrane proteins, which was the largest proteome data set for dental stem cells reported to date. We expect that the SISPROT will be well suited for deep proteome profiling for fewer than 100000 cells and applied for translational studies where multiplexed technology with good label-free quantification precision is required.

SCI ; EI

英语

NI论文

第一 ; 通讯

Shenzhen Innovation of Science and Technology Commission[JCYJ20150901153557178]

Chemistry

Chemistry, Analytical

WOS:000379636600039

AMER CHEMICAL SOC

20162102411177

Biological membranes ; Correlation methods ; Cytology ; Desalination ; Mass spectrometry ; Molecular biology ; Stem cells

Water Treatment Techniques:445.1 ; Biological Materials and Tissue Engineering:461.2 ; Biology:461.9 ; Chemistry:801 ; Organic Compounds:804.1 ; Mathematical Statistics:922.2

CHEMISTRY

Web of Science

1.South Univ Sci & Technol China, Dept Chem, Shenzhen 518055, Peoples R China
2.South Univ Sci & Technol China, Shenzhen Key Lab Cell Microenvironm, Shenzhen 518055, Peoples R China
3.Fudan Univ, Dept Chem, Shanghai 200433, Peoples R China
4.Shenzhen Univ, ENT Inst, Shenzhen Longgang ENT Hosp, Shenzhen 518172, Peoples R China

Chen, Wendong,Wang, Shuai,Adhikari, Subash,et al. Simple and Integrated Spintip-Based Technology Applied for Deep Proteome Profiling[J]. ANALYTICAL CHEMISTRY,2016,88(9):4864-4871.

Chen, Wendong.,Wang, Shuai.,Adhikari, Subash.,Deng, Zuhui.,Wang, Lingjue.,...&Tian, Ruijun.(2016).Simple and Integrated Spintip-Based Technology Applied for Deep Proteome Profiling.ANALYTICAL CHEMISTRY,88(9),4864-4871.

Chen, Wendong,et al."Simple and Integrated Spintip-Based Technology Applied for Deep Proteome Profiling".ANALYTICAL CHEMISTRY 88.9(2016):4864-4871.