Single unit filter-aided method for fast proteomic analysis of tear fluid.

Area of Science:

• Biochemistry
• Proteomics
• Ophthalmology

Background:

• Human tear fluid is a rich source of protein biomarkers for eye diseases like Graves' ophthalmopathy.
• Current Schirmer tear test methods present challenges in sample handling and protein extraction, potentially leading to sample loss.
• Efficient protein extraction is crucial for comprehensive tear proteome analysis.

Purpose of the Study:

• To develop and optimize a novel single-unit filter-aided method for tear sample handling and protein extraction.
• To systematically investigate optimal conditions for protein extraction from Schirmer tear test strips.
• To establish a comprehensive catalogue of the human tear proteome for future biomarker research.

Main Methods:

• Development of a single-unit filter-aided device for integrated tear collection and protein extraction.
• Systematic investigation of various extraction buffer conditions, including ammonium bicarbonate and sodium chloride concentrations.
• Analysis of extracted proteins using one-dimensional and two-dimensional liquid chromatography tandem mass spectrometry (LC-MS/MS).

Main Results:

• Optimized extraction using 100 mM ammonium bicarbonate with 50 mM NaCl yielded the highest number of identified proteins via 1D LC-MS/MS.
• A total of 1526 proteins were identified using the optimized method combined with 2D LC-MS/MS analysis.
• The study successfully generated a comprehensive dataset of the tear proteome.

Conclusions:

• The novel filter-aided method significantly improves protein extraction efficiency from tear samples.
• This optimized approach enhances the study of the tear proteome and facilitates biomarker discovery for eye-related diseases.
• The generated protein dataset serves as a valuable resource for future ophthalmological research.