PepS: An Innovative Microfluidic Device for Bedside Whole Blood Processing before Plasma Proteomics Analyses.

Area of Science:

• Clinical proteomics
• Biomarker analysis
• Mass spectrometry (MS)-based proteomics

Background:

• Immunoassays are standard for protein quantification but have limitations.
• Mass spectrometry (MS) offers a promising alternative for protein biomarker profiling.
• Clinical translation of MS proteomics is hindered by complex, time-consuming sample preparation, especially for plasma's wide dynamic range.

Purpose of the Study:

• To develop an automated microfluidic device for rapid plasma sample preparation.
• To streamline bottom-up MS-based proteomics analysis for clinical applications.
• To address preanalytical challenges in plasma processing for high-throughput biomarker discovery.

Main Methods:

• Design and implementation of a microfluidic device (PepS) for automated sample processing.
• Integration of plasma separation, albumin depletion, protein digestion, and peptide stabilization.
• Operation via a compact instrument with automated fluidics and thermal control.
• Performance evaluation using discovery and targeted proteomics.

Main Results:

• The PepS device successfully automates plasma separation, digestion, and peptide stabilization in under 2 hours.
• Demonstrated detection of three clinical protein biomarkers (ALT1, CRP, myoglobin) using the automated workflow.
• Validated the device's performance in both discovery and targeted proteomics analyses.

Conclusions:

• The PepS microfluidic device significantly simplifies and accelerates plasma sample preparation for MS proteomics.
• This automated solution facilitates the clinical adoption of proteomics for health status monitoring.
• The device has the potential to streamline clinical proteomics studies and biomarker assays.