Single-Color Barcoding for Multiplexed Hydrogel Bead-Based Immunoassays.

Area of Science:

• Biomaterials Science
• Analytical Chemistry
• Microfluidics

Background:

• Precision medicine demands simultaneous detection of multiple biomarkers in complex samples like blood.
• Existing barcoded hydrogel bead immunoassays often involve complex or resource-intensive encoding/decoding methods.

Purpose of the Study:

• To present an efficient, single-step method for fabricating barcoded, functionalized hydrogel beads.
• To develop a simple and versatile barcoding strategy for multiplex bioanalytical processes.

Main Methods:

• Utilized droplet-based microfluidics and photochemically induced C-H insertion reactions.
• Integrated photo-crosslinking, (bio-) functionalization, and barcode integration in a single step.
• Created single-color barcodes using green-fluorescent particles of varying sizes and concentrations for simple readout.

Main Results:

• Successfully fabricated functionalized hydrogel beads with integrated single-color barcodes.
• Demonstrated simple and simultaneous readout using a standard plate reader.
• Validated the bead performance for detecting Lyme disease-specific antibodies in patient sera using a 3x3 matrix.

Conclusions:

• The novel barcoding strategy is simple, versatile, and does not interfere with bioanalytical processes.
• This method offers an attractive solution for multiplex bioanalytical processes in precision medicine.
• The developed hydrogel beads facilitate efficient biomarker detection in complex biological samples.