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Updated: Jun 26, 2026
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Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
Published on: August 17, 2015
Absolute ProGRP quantification in a clinical relevant concentration range using LC-MS/MS and a comprehensive internal
Bjørn Winther1, Petter Moi, Marianne S Nordlund
1Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo, Oslo, Norway.
Summary
This study presents a new liquid chromatography-tandem mass spectrometry method for precisely measuring the small cell lung cancer biomarker ProGRP in human serum. The developed assay achieves low detection limits, aiding in early cancer diagnosis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Oncology
Background:
- Small cell lung cancer (SCLC) requires sensitive biomarkers for early detection.
- ProGRP is a validated biomarker for SCLC, necessitating accurate quantification methods.
Purpose of the Study:
- To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for absolute quantification of ProGRP.
- To establish a reliable assay for ProGRP measurement in human serum.
Main Methods:
- Protein precipitation using acetonitrile followed by tryptic digestion.
- On-line restricted access media (RAM) for sample clean-up and enrichment.
- LC separation on a BioBasic C8 column coupled to a triple quadrupole MS operating in selected reaction monitoring (SRM) mode.
- Use of an acetylated ProGRP internal standard for improved quantification accuracy.
Main Results:
- Achieved a detection limit of 150 pg ProGRP on column in human serum.
- Demonstrated the method's suitability for analyzing realistic clinical samples.
- Internal standard corrected for variations in sample pretreatment and trypsination.
Conclusions:
- The developed LC-MS/MS method provides accurate and sensitive absolute quantification of ProGRP in human serum.
- This assay is a valuable tool for SCLC biomarker monitoring and clinical diagnostics.
- The use of an internal standard enhances the robustness of the quantification process.
