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Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets
Published on: March 17, 2023
Wavelength-scanning surface plasmon resonance imaging for label-free multiplexed protein microarray assay
Soichi Otsuki1, Mitsuru Ishikawa
1Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho, Takamatsu 761-0395, Japan. otsuki-so@aist.go.jp
Biosensors & Bioelectronics
|July 20, 2010
Summary
This study introduces wavelength-scanning surface plasmon resonance (SPR) imaging for label-free protein microarray analysis. The novel method accurately measures molecular binding and layer thickness, achieving high sensitivity for biomolecular interactions.
Area of Science:
- Biophotonics
- Surface Chemistry
- Analytical Chemistry
Background:
- Protein microarrays are crucial for diagnostics and drug discovery.
- Label-free detection methods are needed to simplify assays and reduce costs.
- Surface Plasmon Resonance (SPR) offers label-free detection but can be limited in multiplexing and sensitivity.
Purpose of the Study:
- To develop and validate a wavelength-scanning SPR imaging technique for label-free, multiplexed protein microarray analysis.
- To demonstrate the capability of the method for precise layer thickness estimation.
- To assess the real-time monitoring of biomolecular binding events with high sensitivity.
Main Methods:
- Utilized wavelength-scanning SPR imaging with a gold-coated substrate.
- Implemented dual compensation for wavelength dependence and temporal fluctuations.
- Developed automated SPR wavelength determination per pixel.
- Quantified layer thicknesses and monitored avidin-biotin binding in real-time.
Main Results:
- Successfully estimated thicknesses of thiol, biotin, and spacer layers.
- Demonstrated real-time monitoring of specific avidin-biotin binding.
- Achieved a limit of detection of 20 pm in SPR wavelength (5 pm in thickness).
- Validated the method for label-free, multiplexed protein microarray assays.
Conclusions:
- Wavelength-scanning SPR imaging is a powerful optical tool for label-free, multiplexed protein microarrays.
- The technique enables accurate quantification of molecular layers and binding kinetics.
- This method holds significant potential for high-throughput biomolecular analysis and diagnostics.
