2019
DOI: 10.1021/acs.jproteome.9b00545
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Improved Nanoflow RPLC-CZE-MS/MS System with High Peak Capacity and Sensitivity for Nanogram Bottom-up Proteomics

Abstract: Novel mass spectrometry (MS)-based proteomic tools with extremely high sensitivity and high peak capacity are required for comprehensive characterization of protein molecules in mass-limited samples. We reported a nanoRPLC-CZE-MS/MS system for deep bottom-up proteomics of low micrograms of human cell samples in previous work. In this work, we improved the sensitivity of the nanoRPLC-CZE-MS/MS system drastically via employing bovine serum albumin (BSA)-treated sample vials, improving the nanoRPLC fraction colle… Show more

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Cited by 24 publications
(30 citation statements)
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“…To minimize sample loss due to the adsorption on the inner wall of the insert tube used for CZE-MS, we coated the inner wall with BSA according to our previous work. 36 In total, seven zebrafish brain sections were collected by LCM from optic tectum (Teo, 5 sections) and telencephalon (Tel, 2 sections) of a 20 mm thick brain slice for the analysis, Fig. 2.…”
Section: Development Of An Og-based Sample Preparation Methods For Cz...mentioning
confidence: 99%
See 1 more Smart Citation
“…To minimize sample loss due to the adsorption on the inner wall of the insert tube used for CZE-MS, we coated the inner wall with BSA according to our previous work. 36 In total, seven zebrafish brain sections were collected by LCM from optic tectum (Teo, 5 sections) and telencephalon (Tel, 2 sections) of a 20 mm thick brain slice for the analysis, Fig. 2.…”
Section: Development Of An Og-based Sample Preparation Methods For Cz...mentioning
confidence: 99%
“…Each CZE sample vial was first coated with 2 mg mL À1 bovine serum albumin (BSA) to reduce sample adsorption to the inner walls during analysis. 36 The procedure is as follows:…”
Section: Cze-esi-ms/msmentioning
confidence: 99%
“…At least two different new approaches have been introduced recently in this regard. First, methods were developed allowing analysis with minimum sample requirement, for example, N-glycan analysis from only 100 cells and nanoRPLC-CZE-MS/MS system for proteomics analysis consuming small peptide amounts roughly corresponding to 1000 cells [142,143]. Second, improvements in loading capacity were recently reported with up to 2 μl injections for bottom-up analysis of complex protein samples [144][145][146].…”
Section: Methodological Advancementsmentioning
confidence: 99%
“…We attribute the drastic improvement of proteoform IDs to the high separation efficiency of CZE for proteoforms, [23] high sensitivity of CZE-MS for proteoform detection, [23][24][25] and high orthogonality of LC and CZE for biomolecule separations. [21,26] The features of CZE-MS/MS for TDP have been systematically reviewed recently. [27,28] The list of identified proteoforms is shown in Supporting Information II.…”
Section: Deep Tdp Of Human Colorectal Cancer Cell Lines-pagementioning
confidence: 99%