Data-Independent Acquisition MS Analysis of Peptides

Peptides were analysed in a data‐independent acquisition (DIA) mode by the liquid chromatography (LC) MS, equipped with an EASY‐nanoLC 1200 HPLC system and Orbitrap Fusion Lumos Tribrid mass spectrometer (Thermo) as we previously described with minor modifications (Lu et al., 2019 (link); Tang et al., 2018 ; Zhang et al., 2017 (link)). For LC separation, 1 μg of peptide was loaded into an analytical column (Omics High Resolution, 0.1 mm × 50 cm) and separated with a 130 min gradient and online DIA MS analysis. The DIA MS parameters were set as follows. (1) MS parameters: detector type, orbitrap; orbitrap resolution, 120000; scan range, 350–1200 m/z; RF lens, 30%; AGC target, 1000000; maximum injection time, 100 ms. (2) MS/MS parameters: isolation mode, quadrupole; activation type, HCD; collision energy, 33%; detector type, orbitrap; orbitrap resolution, 50000; mass range, normal; scan range: 200–2000 m/z; AGC target, 100000; maximum injection time, 86 ms. All MS raw data are available in iProX (Ma et al., 2019 ) (accession number: PXD026034).

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Guo J., Wu C., Lin X., Zhou J., Zhang J., Zheng W., Wang T, & Cui Y. (2021). Establishment of a simplified dichotomic size‐exclusion chromatography for isolating extracellular vesicles toward clinical applications. Journal of Extracellular Vesicles, 10(11), e12145.

Publication 2021

EASY‐nanoLC 1200 HPLC system Orbitrap Fusion Lumos Tribrid mass spectrometer

Hplc Isolation Lens Liquid chromatography Ms ms Peptide Scan

Corresponding Organization : Jinan University

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Variable analysis

independent variables

Liquid chromatography (LC) separation method
DIA MS analysis parameters

dependent variables

Peptide identification and quantification

control variables

Experimental parameters (e.g., detector type, resolution, scan range, AGC target, injection time) kept constant across MS and MS/MS analysis

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