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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
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Surface Plasmon Resonance is an Analytically Sensitive Method for Antigen Profiling of Extracellular Vesicles
Elmar L Gool1,2,3, Ivan Stojanovic4, Richard B M Schasfoort4
1Department of Biomedical Engineering and Physics; E.L.Gool@amc.nl.
Clinical Chemistry
|August 9, 2017
Summary
Surface plasmon resonance imaging (SPRi) offers a sensitive method for extracellular vesicle (EV) phenotyping. SPRi detects lower antigen levels than flow cytometry (FCM), correlating well with mean antigen density on EVs.
Area of Science:
- Biotechnology
- Nanotechnology
- Analytical Chemistry
Background:
- Extracellular vesicles (EVs) characterization is challenging due to their small size, low refractive index, and limited surface antigens.
- Accurate EV phenotyping is crucial for understanding their biological roles and developing diagnostics.
Purpose of the Study:
- To evaluate the potential of a 48-multiplex surface plasmon resonance imaging (SPRi) system for extracellular vesicle (EV) phenotyping.
- To compare the performance of SPRi with traditional flow cytometry (FCM) in detecting and quantifying EV surface antigens.
Main Methods:
- Investigated a 48-multiplex SPRi system for EV phenotyping.
- Measured antigen surface density of 11 antigens on human breast cancer cell lines (HS578T, MCF7, SKBR3) and their EVs.
- Compared SPRi results with flow cytometry (FCM) data for both cells and EVs.
Main Results:
- SPRi and FCM signals for antigen exposure correlated well for cells (R² ranging from 0.60 to 0.78).
- SPRi detected significantly more antibody-EV pairs (31/33) compared to FCM (5/33) due to lower background signals.
- SPRi measurements of EV antigen density correlated strongly with parental cell antigen density (R² ranging from 0.49 to 0.77).
Conclusions:
- SPRi responses demonstrate a strong correlation with mean antigen density on extracellular vesicles (EVs).
- SPRi exhibits higher sensitivity than FCM in detecting low antigen-exposure levels on EVs.
- SPRi's ensemble measurement approach provides a more comprehensive analysis of EV surface antigens compared to single-EV detection by FCM.
