Sample-sparing multiplexed antibody Fc biomarker discovery using a reconfigurable integrated microfluidic platform

Area of Science:

• Biomarker Discovery
• Immunology
• Microfluidics

Background:

• Endemic infectious disease control requires sensitive, specific, and accessible biomarkers.
• Antibody fragment crystallizable (Fc) regions, including glycosylation, show promise as pathogen-specific biomarkers.
• Current Fc feature screening methods are often impractical due to high sample volume, cost, and expertise requirements.

Purpose of the Study:

• To develop a simple, flexible, and reconfigurable microfluidic device for high-throughput, multiplexed screening of antibody features.
• To enable discovery of biomarkers targeting both antibody fragment antigen-binding (Fab) and Fc regions.
• To overcome limitations of existing methods for large-scale antibody Fc feature screening.

Main Methods:

• A microfluidic device was fabricated using rapid prototyping techniques, integrating an antigen microarray with reconfigurable microfluidics.
• The device performs highly multiplexed assays (1400 total) measuring 100 antibody Fab and Fc features from a small sample volume (15 μL).
• Features screened include antigen specificity, antibody isotypes, subclasses, N-glycosylation, and Fc receptor binding.

Main Results:

• The device demonstrated cleanroom-free fabrication and ease of use comparable to standard immunoassay platforms.
• Performance was validated against existing methods.
• Biomarker screening for schistosomiasis identified antibody subclass-based biomarkers distinguishing current infection from former infection and endemic controls.

Conclusions:

• The developed microfluidic device offers a simple, efficient, and scalable solution for antibody biomarker discovery.
• It significantly reduces sample volume and complexity compared to traditional methods.
• The device successfully identified novel biomarkers for schistosomiasis, demonstrating its potential for controlling endemic infectious diseases.