Mass spectrometry protocol for tear fluid proteome profiling and biomarker discovery

Area of Science:

• Proteomics
• Biomarker Discovery
• Clinical Diagnostics

Background:

• Tear fluid is a valuable, non-invasive biofluid for disease biomarker discovery.
• Existing proteomics protocols for tear fluid can be time-consuming and prone to contamination.
• Standardization is needed for reliable tear fluid analysis in clinical settings.

Purpose of the Study:

• To develop and validate a rapid and robust protocol for tear fluid proteomics.
• To enable reproducible biomarker discovery for ocular, neurodegenerative, and systemic diseases.
• To optimize protein recovery and minimize contaminants in tear fluid samples.

Main Methods:

• Tear fluid collection using Schirmer strips from human and mouse models.
• Protein digestion, peptide desalting, and solid-phase extraction for sample purification.
• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) with both data-dependent acquisition (DDA) and data-independent acquisition (DIA) strategies.

Main Results:

• A 5-hour workflow that maximizes protein recovery from tear fluid.
• Minimized contaminants, ensuring high-quality proteomic data.
• Demonstrated reproducibility for biomarker discovery applications.

Conclusions:

• The developed protocol offers an efficient and reliable method for tear fluid proteomics.
• This technique facilitates non-invasive biomarker discovery for a range of diseases.
• The protocol supports advancements in personalized medicine through accessible biofluid analysis.