High-Throughput and High-Sensitivity Biomarker Monitoring in Body Fluid by Fast LC SureQuant IS-Targeted...

Area of Science:

• Proteomics
• Biomarker Discovery
• Analytical Chemistry

Background:

• Targeted proteomics assays are crucial for protein and peptide quantification but face limitations in sensitivity and throughput for complex biological samples.
• Current limitations hinder the widespread clinical application of targeted proteomics methods, particularly for low-abundance proteins.

Purpose of the Study:

• To report a novel targeted proteomics assay combining SureQuant internal standard-triggered method with advanced mass spectrometry and liquid chromatography.
• To demonstrate the assay's high sensitivity and throughput for quantifying proteins across a wide dynamic range in complex biological matrices.

Main Methods:

• Utilized the SureQuant internal standard-triggered targeted method.
• Employed a latest-generation mass spectrometer coupled with the EvoSep One liquid chromatography platform.
• Validated the assay using human wound fluid exudates, a complex biological sample.

Main Results:

• Achieved high sensitivity and throughput, processing 100 samples per day.
• Successfully quantified proteins across six orders of magnitude, including low-abundance biomarkers like tumor necrosis factor A and interleukin 1-β.
• Demonstrated robustness in quantifying trace amounts of biomarkers previously inaccessible to targeted proteomics.

Conclusions:

• The developed assay significantly enhances sensitivity and throughput for targeted proteomics.
• This method expands the capabilities for mass spectrometry-based biomarker quantification, particularly for low-abundance analytes.
• The assay holds promise for advancing clinical proteomics and biomarker discovery.