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FRET-based homogeneous immunoassay on a nanoparticle-based photonic crystal
Jin-Hee Han1, Lakshmana Sudheendra, Ian M Kennedy
1Department of Mechanical and Aerospace Engineering, University of California, Davis, CA, 95616, USA.
Analytical and Bioanalytical Chemistry
|May 10, 2015
Summary
This study demonstrates how fluorescence resonance energy transfer (FRET) on a photonic crystal (PC) platform significantly enhances protein immunoassay speed and sensitivity. The novel PC immunoplatform offers rapid, highly sensitive detection for biological assays.
Area of Science:
- Nanotechnology
- Biotechnology
- Analytical Chemistry
Background:
- Protein-based immunoassays are crucial for diagnostics.
- Enhancing immunoassay speed and sensitivity remains a key challenge.
- Photonic crystals (PCs) offer unique optical properties for signal amplification.
Purpose of the Study:
- To evaluate the potential of fluorescence resonance energy transfer (FRET) within a PC nanostructured array.
- To enhance the speed and sensitivity of protein-based immunoassays.
- To demonstrate a FRET-based homogeneous assay on a PC chip.
Main Methods:
- Utilized 40-nm carboxylated particles conjugated with donor-labeled antibodies as FRET donors.
- Employed electrophoresis for particle trapping on the PC array.
- Leveraged PC phase matching for enhanced fluorescent excitation and emission.
- Performed an immunoglobulin G (IgG)-based immunoassay to validate the platform.
Main Results:
- Achieved rabbit-IgG detection limits of 0.001 μg/mL (20 min) and 0.1 μg/mL (1 min).
- Demonstrated sensitivities 1000x and 10x better than a standard 96-well plate reader, respectively.
- Generated standard curves for incubation times of 20 min and 1 min.
Conclusions:
- FRET on a PC immunoplatform is a facile, effective, rapid, and sensitive detection technology.
- The PC array significantly enhances FRET efficiency through phase matching.
- This approach represents a significant advancement in homogeneous immunoassay development.
