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Updated: Jul 18, 2026
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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
Toward a high-throughput approach to quantitative proteomic analysis: expression-dependent protein identification by
T J Griffin1, D K Han, S P Gygi
1Department of Molecular Biotechnology, University of Washington, Seattle, USA. tgriffin@systemsbiology.org
Journal of the American Society for Mass Spectrometry
|January 5, 2002
Summary
This study introduces a new two-step method using isotope-coded affinity tag (ICAT) technology and software for efficient protein quantification. It focuses on identifying differentially expressed proteins, improving proteomic analysis throughput.
Area of Science:
- Proteomics
- Biochemistry
- Mass Spectrometry
Background:
- Isotope-coded affinity tag (ICAT) technology allows for comparative quantitative proteomic analysis.
- Previous ICAT implementations lacked focused identification of differentially expressed proteins.
- Improving sample throughput is crucial for advanced proteomic studies.
Purpose of the Study:
- To develop an improved, high-throughput ICAT method for focused protein quantification.
- To identify proteins with altered abundance between biological samples.
- To enhance the analysis of differentially expressed proteins in disease states.
Main Methods:
- Developed a two-step approach combining automated peptide quantification software with tandem mass spectrometry.
- Utilized microcapillary electrospray ionization time-of-flight mass spectrometry for initial quantification.
- Employed electrospray quadrupole time-of-flight mass spectrometry and database searching for peptide identification.
Main Results:
- Successfully quantified and identified peptides with differential abundance between samples.
- Applied the method to analyze proteins secreted from human prostate epithelial cells, distinguishing between non-tumorigenic and cancerous states.
- Demonstrated the effectiveness of the new approach in identifying differentially expressed proteins.
Conclusions:
- The developed two-step ICAT approach significantly improves sample throughput and focuses on differentially expressed proteins.
- This method enhances the ability to identify and quantify proteins with altered abundance.
- The technology shows promise for applications in cancer research and biomarker discovery.
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