A Multiwell-Based Detection Platform with Integrated PDMS Concentrators for Rapid Multiplexed Enzymatic Assays.

Area of Science:

• Biotechnology
• Analytical Chemistry
• Biomedical Engineering

Background:

• Standard laboratory assays often require lengthy incubation and detection times.
• High-throughput screening demands efficient methods for sample concentration and signal amplification.
• Existing microfluidic integration methods can be complex and substrate-dependent.

Purpose of the Study:

• To develop an integrated system (ISAAC-12) for accelerating biochemical assays using microfluidic concentrators.
• To demonstrate the system's capability for rapid detection of matrix metalloproteinase-9 (MMP-9) expression.
• To present a novel fabrication strategy for microfluidic concentrators compatible with standard multiwell plates.

Main Methods:

• Fabrication of microfluidic concentrator chips by direct printing of a conductive ion-permselective polymer on polydimethylsiloxane (PDMS) channels.
• Integration of the microfluidic concentrator chips with standard 12-well plates via reversible bonding.
• Utilizing electrokinetic preconcentration to accelerate enzymatic reactions and enhance signal intensity.
• Detection of matrix metalloproteinase-9 (MMP-9) expression in MDA-MB-231 breast cancer cell line supernatant.

Main Results:

• The ISAAC-12 system successfully accelerated enzymatic reactions and increased end-point signal intensity.
• Rapid detection of MMP-9 expression was achieved in breast cancer cell culture supernatant.
• The decoupling fabrication strategy allowed easy integration with standard multiwell plates without substrate modification.
• The system demonstrated versatility for multiplexed detection by concentrating enzymes and reaction products simultaneously.

Conclusions:

• The ISAAC-12 system offers a versatile and efficient platform for accelerating high-throughput assays.
• The novel microfluidic concentrator chip fabrication and integration method simplifies assay development.
• This technology has potential applications in rapid diagnostics and drug discovery, particularly for biomarker detection.