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MRM assay for quantitation of complement components in human blood plasma - a feasibility study on multiple sclerosis
Melinda Rezeli1, Akos Végvári, Jan Ottervald
1Division of Clinical Protein Science & Imaging, Biomedical Center, Department of Measurement Technology and Industrial Electrical Engineering, Lund University, Lund, Sweden. melinda.rezeli@elmat.lth.se
Journal of Proteomics
|June 28, 2011
Summary
This study developed a new assay to measure inflammation proteins in blood plasma. The method successfully quantified five key proteins, showing potential for disease research like multiple sclerosis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Immunology
Background:
- Plasma proteins are crucial biomarkers for inflammatory diseases.
- Accurate quantification of these proteins is essential for clinical diagnostics and research.
- Existing methods may have limitations in sensitivity or throughput for certain applications.
Purpose of the Study:
- To develop and validate a multiple reaction monitoring (MRM) assay for quantifying seven inflammation-associated plasma proteins.
- To assess the assay's performance in terms of reproducibility and linear range.
- To apply the developed assay to a pilot cohort of multiple sclerosis patients and controls.
Main Methods:
- Development of a targeted mass spectrometry assay using multiple reaction monitoring (MRM).
- Quantification of proteotypic peptides representing complement components and C-reactive protein.
- Assay validation using depleted plasma digests on a linear ion trap mass spectrometer.
Main Results:
- Successfully quantified 5 out of 7 target inflammation proteins.
- Achieved reasonable reproducibility (RSD ≤ 25%) over a 2-order linear dynamic range.
- Demonstrated assay utility by analyzing plasma samples from multiple sclerosis patients and healthy controls.
Conclusions:
- The developed MRM assay is a viable tool for quantifying inflammation biomarkers in plasma.
- The assay shows promise for application in neurological disease research, specifically multiple sclerosis.
- This proof-of-principle study highlights the potential of targeted proteomics for biomarker discovery and validation.
