A targeted proteomics approach to amyloidosis typing.
Matteo Conti1, Irene Poppi1, Thomas Matulli Cavedagna1
1Clinical Mass Spectrometry, Metropolitan Laboratory AUSL, Bologna, Italy.
Clinical Mass Spectrometry (Del Mar, Calif.)
|August 28, 2024
Summary
A new targeted proteomics method accurately identifies amyloidosis-causing proteins in tissues using mass spectrometry. This approach offers a sensitive and specific diagnostic tool for clinical research, aiding in personalized amyloidosis treatment.
Area of Science:
- Proteomics
- Mass Spectrometry
- Clinical Diagnostics
Background:
- Amyloidosis is a severe condition caused by protein deposits in organs and tissues.
- Accurate protein identification is crucial for diagnosis and personalized treatment.
- Current advanced methods like LC-MS/MS are limited to specialized centers.
Purpose of the Study:
- To develop a targeted proteomics approach for amyloid protein typing.
- To enable analysis using low-resolution mass spectrometry without laser microdissection.
- To determine frequently encountered amyloid proteins (immunoglobulin light chains, transthyretin) and tissue-specific reference proteins.
Main Methods:
- Tissue samples were digested, reduced, alkylated, and trypsinized to create peptide mixtures.
- Peptides were purified by SPE and separated by LC.
- Proteotypic peptides were detected using Multiple Reaction Monitoring (MRM) transitions.
Main Results:
- The method demonstrated high specificity and sensitivity for detecting amyloid protein peptides.
- Limit of detection (LOD) values were in the picomole range for key amyloid proteins in different tissues.
- Ratios of amyloid to tissue-specific proteins correlated with amyloid deposit presence.
Conclusions:
- This targeted proteomics approach provides sensitive and specific discrimination of amyloidosis-affected tissues.
- The method is suitable for clinical research applications.
- It facilitates the identification of amyloid proteins for better patient management.
