VEGF Detection via Simplified FLISA Using a 3D Microfluidic Disk Platform

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Biochemistry

Background:

• Fluorescence-linked immunosorbent assay (FLISA) is a standard quantitative method for detecting biochemical changes via antigen-antibody interactions on well-plate platforms.
• Advancements in microfluidic systems enable the implementation of FLISA on microfluidic disks for high-resolution detection of trace biochemical reactions.

Purpose of the Study:

• To develop a novel microfluidic system for enhanced FLISA performance.
• To reduce reagent consumption and assay time for biochemical detection.

Main Methods:

• A microfluidic disk with a unique three-dimensional incubation chamber was designed and fabricated.
• The system utilizes centrifugal and gravitational forces for automated control of antibody-conjugated microbead movement during incubation.
• Vascular endothelial growth factor (VEGF) was detected sequentially using the 3D microfluidic disk FLISA protocol.

Main Results:

• The 3D microfluidic disk reduced reagent volume by 1/10 and assay time to under one hour.
• Automated microbead movement within the 3D chamber ensured efficient antigen-antibody and substrate binding.
• The system demonstrated effective sequential detection of VEGF concentrations in ng/mL without manual intervention.

Conclusions:

• The proposed 3D microfluidic disk offers a highly efficient and automated platform for FLISA.
• This technology significantly enhances sensitivity and reduces resource requirements for biochemical assays.
• The system holds potential for point-of-care diagnostics and high-throughput screening applications.