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Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
Published on: August 17, 2015
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LC-SRM-Based Targeted Quantification of Urinary Protein Biomarkers
Yuqian Gao1, Hui Wang1, Carrie D Nicora1
1Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2017
Summary
Liquid chromatography-selected reaction monitoring (LC-SRM) offers sensitive and reproducible protein quantification. This method accurately measures protein biomarkers in human urine using stable isotope-labeled standards.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Proteomics
Background:
- Liquid chromatography-selected reaction monitoring (LC-SRM) is a highly sensitive, reproducible, and multiplexing technique for protein quantification.
- Accurate measurement of targeted proteins in complex biological samples is crucial for biomarker discovery.
- Stable heavy isotope-labeled peptides serve as effective internal standards for precise quantification.
Purpose of the Study:
- To provide a detailed protocol for applying LC-SRM.
- To enable the quantification of candidate protein biomarkers in human urine.
Main Methods:
- Utilizing liquid chromatography coupled with selected reaction monitoring (LC-SRM).
- Employing stable heavy isotope-labeled peptides as internal standards.
- Quantifying surrogate proteotypic peptides for targeted proteins in human urine samples.
Main Results:
- Demonstrated the successful application of LC-SRM for protein biomarker quantification.
- Established a step-by-step protocol for reproducible results.
- Validated the accuracy of LC-SRM in complex human urine matrices.
Conclusions:
- LC-SRM is a powerful and accurate method for quantifying protein biomarkers in human urine.
- The developed protocol facilitates the application of LC-SRM in clinical and research settings.
- This technique supports biomarker discovery and validation efforts.
