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Determination of exosome concentration in solution using surface plasmon resonance spectroscopy
Déborah L M Rupert1, Cecilia Lässer, Maria Eldh
1Department of Applied Physics, Chalmers University of Technology , SE-412 96 Gothenburg, Sweden.
Analytical Chemistry
|May 23, 2014
Summary
This study presents a label-free surface plasmon resonance (SPR) method for quantifying exosomes. The technique measures exosome binding rates to determine their concentration in biological fluids with improved accuracy.
Area of Science:
- Biotechnology
- Nanotechnology
- Analytical Chemistry
Background:
- Exosomes are crucial for intercellular communication and hold potential for diagnostics and therapeutics.
- Accurate quantification of exosomes in biological fluids is essential for their clinical applications.
- Current methods for exosome detection and quantification face challenges in sensitivity and specificity.
Purpose of the Study:
- To develop and validate a label-free surface-based sensing method using surface plasmon resonance (SPR) for exosome concentration determination.
- To analyze human mast cell secreted exosomes (CD63+) using SPR.
- To establish a reliable quantification method accounting for exosome characteristics and SPR limitations.
Main Methods:
- Utilized surface plasmon resonance (SPR) with anti-CD63 antibody functionalized sensors.
- Measured the diffusion-limited binding rate of exosomes to the sensor surface.
- Converted SPR response to surface-bound mass and related it to exosome concentration.
- Employed calibration with proteins and lipid vesicles, considering exosome size distribution, SPR probing depth, and adsorption-induced deformation.
Main Results:
- Successfully quantified exosomes by measuring their binding kinetics via SPR.
- Established a method relating SPR-derived mass uptake over time to exosome concentration.
- Achieved an estimated accuracy of better than ±50% for concentration determination, with potential for improvement.
Conclusions:
- The developed SPR-based method offers a sensitive, label-free approach for exosome quantification in complex biological samples.
- The method accounts for critical factors influencing SPR measurements of exosomes, enhancing reliability.
- This technique supports the advancement of exosome-based diagnostics and therapeutic delivery systems.
