Towards reproducible MRM based biomarker discovery using dried blood spots

Area of Science:

• Biochemistry and Molecular Biology
• Analytical Chemistry
• Clinical Proteomics

Background:

• Growing interest in dried blood spot (DBS) sampling and multiple reaction monitoring (MRM) for proteomics.
• Previous studies focused on protein detection in DBS, but reproducibility and suitability for high-throughput biomarker discovery remain unclear.

Purpose of the Study:

• To assess the reproducibility of multiplexed targeted protein measurements in DBS compared to serum.
• To evaluate the utility of DBS for high-throughput biomarker discovery in proteomics.

Main Methods:

• Monitoring of 82 medium to high abundance proteins in technical and biological replicates using DBS and serum.
• Application of statistical quality control approaches to detect inaccurate transitions.
• Comparison of protein quantification data between DBS and serum samples.

Main Results:

• Median coefficient of variation (CV) for detected peptides was 13.2% in DBS and 8.8% in serum after statistical quality control.
• A strong correlation (r=0.72) was observed between relative peptide abundance in DBS and serum.
• Demonstrated feasibility of multiplexed targeted protein quantification in DBS.

Conclusions:

• DBS sampling provides reproducible multiplexed targeted protein measurements comparable to serum.
• The combination of DBS and mass spectrometry (MS) is suitable for high-throughput proteomic studies and biomarker discovery.
• This approach has the potential to transform clinical proteomics and personalized medicine through large-scale studies.