Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase
Daniela Waas1, Marc Juraschitz2, Yen-Fu Adam Chen1
1Eximmium Biotechnologies GmbH, Munich, Germany.
Journal of Extracellular Vesicles
|July 31, 2025
Summary
Researchers developed horseradish peroxidase (HRP)-expressing extracellular vesicles (EVs) as reference standards. These HRP+ EVs enable sensitive quantification and standardization of EV analysis for reliable biomarker applications.
Area of Science:
- Biotechnology
- Biomarker Discovery
- Nanomedicine
Background:
- Extracellular vesicles (EVs) are promising circulating biomarkers for diagnostics and therapeutics.
- Current EV research faces challenges in result replication and interpretation due to diverse protocols.
- Lack of qualified reference standards hinders quantitative EV assessment and quality control.
Purpose of the Study:
- To develop a novel class of reference extracellular vesicles (EVs) for standardization.
- To facilitate reliable EV tracing during isolation and purification.
- To enable quantitative assessment and quality control in EV-based diagnostics.
Main Methods:
- Generation of engineered EVs expressing horseradish peroxidase (HRP).
- Quantification of HRP+ EVs via direct HRP activity measurement.
- Quantification of HRP+ EVs via indirect immunodetection of surface HRP.
Main Results:
- HRP+ EVs provide a sensitive and reliable method for EV quantification.
- Demonstrated ability to quantify absolute EV numbers in biological samples.
- Established a method for normalizing clinical specimens in liquid biopsies using HRP+ EVs.
Conclusions:
- HRP-expressing EVs serve as valuable reference standards for EV research.
- This approach enhances the reproducibility and comparability of EV analysis.
- Facilitates the clinical translation of EV-based liquid biopsies.
