Paper-Based Antibody Detection Devices Using Bioluminescent BRET-Switching Sensor Proteins.

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Point-of-Care Diagnostics

Background:

• Microfluidic paper-based analytical devices (microPADs) offer potential for low-cost diagnostics.
• Existing microPADs often require complex sample handling and reagents.
• Novel detection mechanisms are needed for user-friendly, reagent-free operation.

Purpose of the Study:

• To develop fully integrated "sample-in-signal-out" microPADs.
• To utilize bioluminescence resonance energy transfer (BRET) for analyte recognition and signal generation.
• To enable reagent-free, sample volume-independent operation for point-of-care testing.

Main Methods:

• Integration of BRET-based antibody sensing proteins into vertically assembled paper layers.
• Design of devices for on-device blood plasma separation.
• User operation involving a single sample drop and image acquisition via a camera.

Main Results:

• Demonstrated fully integrated, reagent-free microPADs.
• Achieved simultaneous detection of three antibodies (anti-HIV1, anti-HA, anti-DEN1) in whole blood.
• Device operation is independent of sample volume and requires minimal user intervention.

Conclusions:

• The developed microPADs are simple, user-friendly, and suitable for point-of-care testing.
• The BRET-based approach enables sensitive and specific antibody detection.
• These devices are particularly well-suited for low-resource environments.