Capillary zone electrophoresis-mass spectrometry with microliter-scale loading capacity, 140 min separation window and high peak capacity for bottom-up proteomics

The Analyst
Daoyang ChenLiangliang Sun

Abstract

Better peptide separation is required for bottom-up proteomics for further improving the proteome coverage. The two-dimensional liquid chromatography (2D-LC) systems only explore differences among peptides in their hydrophobicity (reversed-phase, RP) and charge (strong cation/anion exchange, SCX/SAX). Alternative separation techniques with different separation mechanisms are required to further improve the separation. Capillary zone electrophoresis (CZE) is an attractive alternative because it has high efficiency for separation of biomolecules and it separates analytes based on their size-to-charge ratios, complementary with LC. However, the low loading capacity and narrow separation window of CZE limit its wide application for large-scale proteomics. In this manuscript, we present an automated CZE-mass spectrometry (MS) system for solving those issues. The CZE-MS system can approach at least half-a-microliter loading capacity with good robustness and reproducibility, can routinely use over 12% of the available sample in the sample vial for analysis, and can generate a 140 min separation window and high peak capacity (∼380) for complex proteome analysis. The results represent the highest peak capacity and the widest separation ...Continue Reading

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Citations

Nov 18, 2017·Journal of the American Society for Mass Spectrometry·Sam B ChoiPeter Nemes
Aug 4, 2018·Electrophoresis·Andrea ŠlampováPetr Gebauer
Sep 22, 2018·Journal of Separation Science·Ivan Mikšík
May 3, 2019·Proteomics·Leah V SchafferLloyd M Smith
Oct 15, 2019·Analytical Methods : Advancing Methods and Applications·Elijah N McCoolLiangliang Sun
Mar 25, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Camille Lombard-Banek, John E Schiel
Feb 26, 2021·Analytical Methods : Advancing Methods and Applications·Zhichang Yang, Liangliang Sun
Aug 12, 2021·Analytical Methods : Advancing Methods and Applications·Bo LiuHong-Xu Chen

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