title: Self-packed core shell nano liquid chromatography columns and silica-based monolithic trap columns for targeted proteomics.
abstract: Self-preparation of nano liquid chromatography (nLC) columns has advantages regarding cost and flexibility. For targeted proteomics, we evaluated several approaches for particle-packing nLC columns and manufacturing fritless silica-based monolithic trap columns (50μm inner diameter). Our preferred approach for nLC column preparation was to magnetically stir Accucore core shell particles (C18 stationary phase) in ACN/water (80/20, v/v) suspensions during pressure-driven filling of polymer-fritted standard fused silica capillaries. The columns were ready for use about one hour after preparation had begun. They had comparable peak capacities (peptides) to commercial columns, and satisfactory within/between-column retention time repeatability, suited for targeted proteomics. Packing with commercial capillary housings/nanospray emitters did not improve performance compared to packing with in-house fritted stock fused silica capillary tubing. For trap columns, several recipes for narrow bore silica-based monolithic columns were evaluated, and we found the recipe by Zou et al. (2005) to be reproducible. Compared to the standard C18 trap column for Accucore nLC columns, monolith trap columns (C8 stationary phase) significantly reduced peak widths. The readily prepared in-house columns were used for targeted detection of the enzyme CYP27A1 in cancer cells, which is associated with proliferation and metastasis of breast cancer.
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Henriette Sjaanes
authors.initials: HS
authors.lastname: Berg
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Kristina Erikstad
authors.initials: KE
authors.lastname: Seterdal
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Tone
authors.initials: T
authors.lastname: Smetop
authors.affiliation: Faculty of Chemistry, University of Latvia, Jelgavas iela 1, Post Box LV-1004, Rīga, Latvia.
authors.forename: Rūdolfs
authors.initials: R
authors.lastname: Rozenvalds
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Ole Kristian
authors.initials: OK
authors.lastname: Brandtzaeg
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Tore
authors.initials: T
authors.lastname: Vehus
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Elsa
authors.initials: E
authors.lastname: Lundanes
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway. Electronic address: stevenw@kjemi.uio.no.
authors.forename: Steven Ray
authors.initials: SR
authors.lastname: Wilson
mesh_terms: D045744:Cell Line, Tumor; D053493:Cholestanetriol 26-Monooxygenase / Q000032:analysis / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000032:analysis* / Q000302:isolation & purification; D011312:Pressure; D040901:Proteomics / Q000379:methods*; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D010455:Peptides; D012822:Silicon Dioxide; C506831:CYP27A1 protein, human; D053493:Cholestanetriol 26-Monooxygenase
keywords: CYP27A1; Column packing; Core shell particles; Liquid chromatography; Nano; Silica monolith; Targeted proteomics