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A Guided Materials Screening Approach for Developing Quantitative Sol-gel Derived Protein Microarrays
Published on: August 26, 2013
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Ag@SiO2-entrapped hydrogel microarray: a new platform for a metal-enhanced fluorescence-based protein assay
Eunji Jang1, Minsu Kim, Won-Gun Koh
1Department of Chemical and Biomolecular Engineering, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-749, South Korea. wongun@yonsei.ac.kr.
The Analyst
|April 4, 2015
Summary
We created a novel hydrogel microarray bioassay using metal-enhanced fluorescence (MEF) for sensitive glucose detection. This platform significantly improves signal detection for diagnostic applications.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Materials Science
Background:
- Protein-based bioassays are crucial for diagnostics.
- Metal-enhanced fluorescence (MEF) offers signal amplification potential.
- Hydrogel microarrays provide a stable platform for enzyme immobilization.
Purpose of the Study:
- To develop a novel protein-based bioassay platform using MEF.
- To detect glucose concentrations using a hydrogel microarray entrapping silica-coated silver nanoparticles (Ag@SiO2).
- To optimize MEF for enhanced fluorescence and improved glucose detection sensitivity.
Main Methods:
- Fabrication of poly(ethylene glycol) (PEG) hydrogel microarrays via photopatterning.
- Immobilization of glucose oxidase (GOX) and peroxidase (POD) enzymes within the hydrogel.
- Incorporation of silica-coated silver nanoparticles (Ag@SiO2) to enhance fluorescence.
- Detection of glucose using a sequential bienzymatic reaction and fluorescence measurement.
Main Results:
- Successful entrapment of enzymes and Ag@SiO2 within hydrogel microarrays without leaching or deactivation.
- Demonstrated fluorescence signal enhancement dependent on silica shell thickness and Ag@SiO2 concentration.
- Achieved optimal MEF with a 17.5 nm silica shell and 0.5 mg mL(-1) Ag@SiO2.
- Observed over a 4-fold fluorescence enhancement for glucose detection (10(-3) mM to 10.0 mM) compared to assays without Ag@SiO2.
- Significantly improved sensitivity and limit of detection (LOD) for glucose measurement.
Conclusions:
- The developed hydrogel microarray platform effectively utilizes MEF for enhanced bioassays.
- The system demonstrates superior sensitivity and LOD for glucose detection.
- This platform is a promising step towards creating MEF-based micro-total-analysis-systems (μ-TAS) when integrated with microfluidics.
