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Peptide Identification Using Tandem Mass Spectrometry

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Updated: Sep 21, 2025

Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry

Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry

Published on: October 24, 2018

Monolithic Materials-Based RPLC-MS for Proteoform Separation and Identification.

Yu Liang¹, Lihua Zhang², Yukui Zhang¹

¹CAS Key Lab of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.

Methods in Molecular Biology (Clifton, N.J.)

|June 3, 2022

Summary

This summary is machine-generated.

New functionalized hybrid monoliths enable high-performance proteoform separation for top-down proteomics. This method enhances proteomic analysis by combining chromatography with mass spectrometry for better identification.

Keywords:

Bridged hybrid monolith Monolithic materials Proteoform separation Top-down proteomic analysis

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Area of Science:

Proteomics
Analytical Chemistry
Materials Science

Background:

Proteoform separation is crucial for complex proteome analysis in top-down proteomics.
Existing methods face challenges due to the high complexity of proteomes.

Purpose of the Study:

To develop functionalized ethylene-bridged hybrid monolithic materials for enhanced proteoform separation.
To integrate these materials with high-resolution mass spectrometry for top-down proteomic analysis.

Main Methods:

Preparation of functionalized ethylene-bridged hybrid monoliths.
Reversed-phase liquid chromatography (RPLC) using the developed monoliths.
Online coupling of RPLC with high-resolution mass spectrometry (HRMS).

Main Results:

The developed monoliths exhibit homogenous functional groups, good chemical stability, and high permeability.
Achieved high-resolution separation, good reproducibility, and low backpressure for proteoforms.
Demonstrated successful online combination with HRMS for proteoform identification.

Conclusions:

Functionalized hybrid monoliths are effective for high-performance proteoform separation in top-down proteomics.
The developed method offers an improved approach for analyzing complex proteomes.
This technique facilitates detailed proteoform separation and identification.