Multiplexed fluorescent microarray for human salivary protein analysis using polymer microspheres and fiber-optic...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Immunotechnology

Background:

• Multiplex protein analysis is crucial for diagnostics.
• Existing methods face challenges in sensitivity and throughput.
• Saliva offers a non-invasive source for biomarkers.

Purpose of the Study:

• To develop and validate a novel fiber-optic microsphere-based immunoassay.
• To enable simultaneous measurement of multiple proteins in saliva.
• To establish a sensitive and efficient platform for salivary diagnostics.

Main Methods:

• Utilized encoded polymer microspheres with distinct fluorescent signatures.
• Employed EDC/NHS chemistry for antibody immobilization on microspheres.
• Assembled a fiber-optic bundle microarray for random microsphere deposition.
• Performed sandwich immunoassays with fluorescent detection (R-phycoerythrin).
• Analyzed data using fluorescence microscopy and custom MATLAB algorithms.

Main Results:

• Successfully developed a protocol for simultaneous measurement of six saliva proteins.
• Demonstrated the capability of the microsphere immunoassay for protein capture and detection.
• Achieved sensitive detection using a sandwich immunoassay format.
• Validated the use of fiber-optic bundles as carriers and imaging platforms.

Conclusions:

• The fiber-optic microsphere immunoassay is a viable method for multiplexed saliva protein analysis.
• This technology offers a sensitive and efficient platform for salivary biomarker discovery and diagnostics.
• The protocol combines advantages of microsphere arrays and fluorescence microscopy for robust immunoassays.