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Published on: January 22, 2019
Hydrogel-Framed Nanofiber Matrix Integrated with a Microfluidic Device for Fluorescence Detection of Matrix
1Department of Chemical and Biomolecular Engineering, Yonsei University , 50 Yonsei-ro, Seodaemun-gu, Seoul 120-749, South Korea.
Abstract:
Matrix metalloproteinases (MMPs) play a pivotal role in regulating the composition of the extracellular matrix and have a critical role in vascular disease, cancer progression, and bone disorders. This paper describes the design and fabrication of a microdevice as a new platform for highly sensitive MMP-9 detection. In this sensing platform, fluorescein isocyanate (FITC)-labeled MMP-9 specific peptides were covalently immobilized on an electrospun nanofiber matrix to utilize an enzymatic cleavage strategy. Prior to peptide immobilization, the nanofiber matrix was incorporated into hydrogel micropatterns for easy size control and handling of the nanofiber matrix. The resultant hydrogel-framed nanofiber matrix immobilizing the peptides was inserted into microfluidic devices consisting of reaction chambers and detection zones. The immobilized peptides were reacted with the MMP-9-containing solution in a reaction chamber, which resulted in the cleavage of the FITC-containing peptide fragments and subsequently generated fluorescent flow at the detection zone. As higher concentrations of the MMP-9 solution were introduced or larger peptide-immobilizing nanofiber areas were used, more peptides were cleaved, and a stronger fluorescence signal was observed. Due to the huge surface area of the nanofiber and small dimensions of the microsystem, a faster response time (30 min) and lower detection limit (10 pM) could be achieved in this study. The hydrogel-framed nanofiber matrix is disposable and can be replaced with new ones immobilizing either the same or different biomolecules for various bioassays, while the microfluidic system can be continuously reused.
Insights
This study presents a novel microdevice for highly sensitive detection of matrix metalloproteinase-9 (MMP-9). The platform uses enzyme-cleaved peptides on nanofibers, achieving a low detection limit for improved diagnostics.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Biochemistry
Background:
- Matrix metalloproteinases (MMPs) are crucial in extracellular matrix regulation and implicated in vascular disease, cancer, and bone disorders.
- Accurate and sensitive detection of MMPs, particularly MMP-9, is vital for disease diagnosis and monitoring.
- Existing detection methods may lack sensitivity, speed, or require complex sample preparation.
Purpose of the Study:
- To design and fabricate a novel microdevice for highly sensitive and rapid detection of MMP-9.
- To develop a sensing platform utilizing immobilized peptides and an enzymatic cleavage strategy for fluorescence-based detection.
- To evaluate the performance of the microdevice in terms of sensitivity, response time, and reusability.
Main Methods:
- Covalent immobilization of fluorescein isocyanate (FITC)-labeled MMP-9 specific peptides onto an electrospun nanofiber matrix.
- Incorporation of the nanofiber matrix into hydrogel micropatterns for enhanced handling and size control.
- Integration of the hydrogel-framed nanofiber matrix into a microfluidic device with reaction and detection zones.
- Detection of MMP-9 based on the enzymatic cleavage of immobilized peptides, generating a measurable fluorescent signal.
Main Results:
- The microdevice achieved a low detection limit of 10 pM for MMP-9.
- A rapid response time of 30 minutes was observed due to the high surface area of nanofibers and microdevice dimensions.
- Fluorescence signal intensity correlated directly with MMP-9 concentration and the area of peptide immobilization.
- The hydrogel-framed nanofiber matrix is disposable, while the microfluidic system is reusable.
Conclusions:
- The developed microdevice offers a highly sensitive and rapid platform for MMP-9 detection.
- The combination of hydrogel micropatterns, electrospun nanofibers, and microfluidics enables efficient bioassays.
- This disposable sensing matrix and reusable microfluidic system present a versatile platform for various bioanalytical applications.
