Applications of Monolithic Column and Isotope Dimethylation by Fangjun Wang

By Fangjun Wang

In this thesis, the writer outlines the advance of recent monolithic columns and isotope dimethyl labeling concepts and their functions in high-performance proteome analyses. notwithstanding kinds of monolithic columns were broadly constructed for chromatography and electrophoresis separation, their program in proteomics for complicated peptide combos separation remains to be a problem. the writer discusses the training of recent monolithic columns and optimization of chromatography separation power to enhance insurance and accuracy of proteome research. additional, the writer describes a unique on-line multidimensional chromatography process mixed with computerized on-line isotope labeling, which considerably improves the throughput, sensitivity and accuracy of quantitative proteomics. as well as the advance of recent applied sciences, the writer investigates the proteome and phosphoproteome expression alterations of scientific hepatocellular carcinoma tissues and the hippocampi of mice with Alzheimer’s illness. The paintings during this thesis has ended in a number of courses in high-profile journals within the fields of analytical chemistry and proteome research.

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Wang F, Jiang X, Feng S, Tian R, Han G, Liu H, Ye M, Zou H (2007) Automated injection of uncleaned samples using a ten-port switching valve and a strong cation-exchange trap column for proteome analysis. Fu H, Xie C, Dong J, Huang X, Zou H (2004) Monolithic column with zwitterionic stationary phase for capillary electrochromatography. Dong J, Zhou H, Wu R, Ye M, Zou H (2007) Specific capture of phosphopeptides by Zr4+modified monolithic capillary column. Wang F, Dong J, Jiang X, Ye M, Zou H (2007) Capillary trap column with strong cationexchange monolith for automated shotgun proteome analysis.

The solution from the monolithic column flowed into a 500 nL 1,100-DAD micro cell UV detector (Agilent). 1 % FA and 10 % acetonitrile was used to estimate the void time of this system at the same flow rate because benzonic acid with considerable UV absorption is neutral in acidic solution.  × 2 cm was also measured following the same procedures. 1. This system was used to systematically study the influence of dead volume between trap and analytical columns on the separation performance and proteomic coverage.

38]. Copyright 2007 American Chemical Society aViscosity data were from Ref. [34] bPermeability can be calculated by Darcy’s Law, k = ηLF/(πr2△P), where η is the viscosity, L is the column length, F is the solvent flow rate, r is the radius of trap column, and △P is the column back pressure cThe packed length of particulate trap columns was kept at 1 cm due to the pressure limit at high flow rate 20 2 Online Sample Injection and Multidimensional Chromatography porogenic solvent. Monolithic column used in proteome analysis should be stable enough in different separation buffers and not exhibit extra swelling or shrinking, which always influences the permeability of monolith.

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