title: Intact-protein trapping columns for proteomic analysis in capillary high-performance liquid chromatography.
abstract: A new type of monolithic trapping columns with high mechanical strength was prepared by thin-layer sol-gel coating method and applied to trapping intact proteins for on-line capillary liquid chromatography. Monolithic trapping columns were fabricated by entrapping C8 reversed-phase particles into the capillary columns through a sol-gel network, which was formed by hydrolysis and polycondensation of methyltriethoxysilane. Hundreds times of trapping/untrapping for intact proteins were carried out. The trapping columns showed long-term stability up to 300 bar. Recovery, loading capacity and reproducibility of trapping columns were evaluated using four proteins. The recovery of four protein mixtures for the C8 monolithic trapping columns was 99.3% on average. The loading capacity of 5 mm × 320 μm i.d. C8 trapping columns for the protein mixtures was 30 μg. Day-to-day relative standard deviation (RSD) values for recoveries of protein mixtures on the same C8 trapping column ranged from 2.34 to 5.87%, column-to-column RSD values were from 3.01 to 6.81%. The C8 trapping columns were used to trap normal mouse liver intact proteins in a capillary liquid chromatography system. Results demonstrated high efficiency of the monolithic trapping columns for trapping intact proteins for proteomic analysis in on-line capillary liquid chromatography system.
authors.affiliation: Department of Chemistry, Fudan University, Shanghai 200433, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Guan
authors.forename: Guoquan
authors.initials: G
authors.lastname: Yan
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gao
authors.forename: Guangfeng
authors.initials: G
authors.lastname: Hong
authors.forename: Chunhui
authors.initials: C
authors.lastname: Deng
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000085:Acetates / Q000737:chemistry; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D005782:Gels; D008110:Liver Extracts / Q000737:chemistry; D051379:Mice; D008855:Microscopy, Electron, Scanning; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D015203:Reproducibility of Results; D012965:Sodium Chloride / Q000737:chemistry
chemical_list: D000085:Acetates; D005782:Gels; D008110:Liver Extracts; D011506:Proteins; D012965:Sodium Chloride; C018824:ammonium acetate