title: Rapid fabrication of functionalized plates for peptides, glycopeptides and protein purification and mass spectrometry analysis.
abstract: A rapid and simple approach for fabricating a disposable functionalized membrane on matrix-assisted laser desorption ionization (MALDI) targets, glass, or plastic substrates, without using complex mechanical protocols or chemical reactions, was developed for sample enrichment and mass spectrometry analysis. By coating functionalized-silica particles on a polydimethylsiloxane (PDMS)-coated plate, these particles can form a monolayer of materials on the PDMS membrane for sample handling without peeling off. An octadecyl(C18)-functionalized plate was fabricated by coating porous C18-silica particles on a PDMS-coated plate. The C18 particle-coated PDMS plate (CP plate) has better sensitivity than C18 tips and magnetic nanoparticles, along with a higher sample recovery (64.3 ± 4.9%) compared to the C18 tip method, when analyzing trace amounts of 5 fm BSA digest samples. The CP plate shows significantly higher urea/SDS removal efficiency on the cell lysate proteome compared to C18 tips. The capacity of the C18 spot (∼2.8 mm in diameter) on the CP plate was ∼10 μg of BSA digests. A hydrophilic particle-coated PDMS plate was also fabricated and successfully used for glycopeptide enrichment and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis.
authors.affiliation: Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung 40402, Taiwan. cjchen@mail.cmu.edu.
authors.forename: Hsin-Yi
authors.initials: HY
authors.lastname: Liao
authors.affiliation: Department of Medical Genetics and Medical Research, China Medical University Hospital, Taichung 40402, Taiwan.
authors.forename: Fuu-Jen
authors.initials: FJ
authors.lastname: Tsai
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan.
authors.forename: Chien-Chen
authors.initials: CC
authors.lastname: Lai
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Mei-Chun
authors.initials: MC
authors.lastname: Tseng
authors.affiliation: Graduate Institute of Clinical Medical Science, China Medical University, Taichung 40402, Taiwan.
authors.forename: Chung Y
authors.initials: CY
authors.lastname: Hsu
authors.affiliation: Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung 40402, Taiwan. cjchen@mail.cmu.edu and Graduate Institute of Integrated Medicine, China Medical University, Taichung 40402, Taiwan.
authors.forename: Chao-Jung
authors.initials: CJ
authors.lastname: Chen
mesh_terms: D004129:Dimethylpolysiloxanes / Q000737:chemistry; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics; D012822:Silicon Dioxide / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors
chemical_list: D004129:Dimethylpolysiloxanes; D006020:Glycopeptides; D011506:Proteins; C013830:baysilon; D012822:Silicon Dioxide