Related Experiment Video
Updated: Jun 7, 2026
10:37
Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
Clinical proteomics: liquid chromatography-mass spectrometry purification systems
1National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland.
Methods in Molecular Biology (Clifton, N.J.)
|October 28, 2010
Summary
Liquid chromatography/mass spectrometry (LC/MS) is a powerful tool for clinical proteomics, aiding protein identification and biomarker discovery. This chapter details simple protocols for peptide analysis using one-dimensional and two-dimensional LC/MS techniques.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Liquid chromatography/mass spectrometry (LC/MS) is a standard technology in clinical proteomic research.
- It is essential for protein identification, characterization, and biomarker discovery.
Purpose of the Study:
- To describe simple protocols for protein digestion.
- To detail peptide analysis methods using LC/MS.
Main Methods:
- Protein digestion protocols.
- One-dimensional reversed-phase liquid chromatography (1D-RP-LC).
- Two-dimensional liquid chromatography (2D-LC), including strong cation exchange (SCX) followed by reversed-phase chromatography (RP).
- Tandem mass spectrometry (MS/MS) for peptide analysis.
Main Results:
- The chapter provides straightforward methods for preparing and analyzing digested peptides.
- Demonstrates the application of both 1D-LC and 2D-LC coupled with MS/MS.
Conclusions:
- The described protocols facilitate routine protein identification and characterization in clinical proteomic studies.
- These methods support the discovery of protein biomarkers using LC/MS technology.
Related Concept Videos
01:33Peptide Identification Using Tandem Mass Spectrometry
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
01:21Mass Spectrometry: Complex Analysis
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...