title: Functionalization of hybrid monolithic columns via thiol-ene click reaction for proteomics analysis.
abstract: The vinyl-functionalized hybrid monolithic columns (75 and 150μm i.d.) were prepared via sol-gel chemistry of tetramethoxysilane (TMOS) and vinyltrimethoxysilane (VTMS). The content of accessible vinyl groups was further improved after the monolithic column was post-treated with vinyldimethylethoxysilane (VDMES). The surface properties of monolithic columns were tailored via thiol-ene click reaction by using 1-octadecanethiol, sodium 3-mercapto-1-propanesulfonate and 2,2'-(ethylenedioxy)diethanethiol/vinylphosphonic acid, respectively. The preparing octadecyl-functionalized monolithic columns were adopted for proteomics analysis in cLC-MS/MS. A 37-cm-long×75-μm-i.d. monolithic column could identify 3918 unique peptides and 1067 unique proteins in the tryptic digest of proteins from HeLa cells. When a 90-cm-long×75-μm-i.d. monolithic column was used, the numbers of unique peptides and proteins were increased by 82% and 32%, respectively. Furthermore, strong cation exchange (SCX) monolithic columns (4cm in length×150μm i.d.) were also prepared and coupled with the 37-cm-long×75-μm-i.d. octadecyl-functionalized monolithic column for two-dimensional SCX-RPLC-MS/MS analysis, which could identify 17114 unique peptides and 3211 unique proteins.
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Zhongshan
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Zheyi
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Wang
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China. Electronic address: junjieou@dicp.ac.cn.
authors.forename: Junjie
authors.initials: J
authors.lastname: Ou
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China. Electronic address: mingliang@dicp.ac.cn.
authors.forename: Mingliang
authors.initials: M
authors.lastname: Ye
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Hanfa
authors.initials: H
authors.lastname: Zou
mesh_terms: D002853:Chromatography, Liquid; D057930:Click Chemistry; D006367:HeLa Cells; D006801:Humans; D063065:Organophosphonates / Q000737:chemistry; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D012821:Silanes / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014753:Vinyl Compounds / Q000737:chemistry
chemical_list: C579787:3-mercapto-1-propanesulfonate; D063065:Organophosphonates; D010455:Peptides; D011506:Proteins; D012821:Silanes; D013438:Sulfhydryl Compounds; D014753:Vinyl Compounds; C529208:trimethoxyvinylsilane; C433467:vinylphosphonic acid; C055360:n-octadecyl mercaptan; C061205:tetramethoxysilane; D012822:Silicon Dioxide
keywords: Liquid chromatography; Monolithic column; Proteomics analysis; Thiol-ene click reaction; Two-dimensional separation