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Recombinant extracellular vesicles as biological reference material for method development, data normalization and
Edward Geeurickx1,2, Lien Lippens1,2,3, Pekka Rappu4
1Laboratory of Experimental Cancer Research, Department of Human Structure and Repair, Ghent University, Ghent, Belgium.
Nature Protocols
|January 16, 2021
Summary
Researchers developed recombinant extracellular vesicles (rEV) as a reference material for extracellular vesicle (EV) analysis. This standardized approach enhances the reliability and reproducibility of EV-based diagnostics and therapeutics.
Area of Science:
- Biotechnology
- Biomedical Engineering
- Molecular Biology
Background:
- Extracellular vesicles (EVs) show promise for diagnostic and therapeutic applications.
- Standardized reference materials are crucial for reliable EV analysis and data interpretation.
- Existing methods lack robust controls for EV quantification and characterization.
Purpose of the Study:
- To engineer recombinant extracellular vesicles (rEV) as a biological reference material.
- To provide a protocol for rEV production, quality assurance, and application in EV analysis.
- To demonstrate the utility of rEV for method development, data normalization, and assessment of pre-analytical variables.
Main Methods:
- Engineered recombinant EVs (rEV) with biochemical and biophysical similarity to native EVs.
- Developed a protocol for rEV production, separation, and quality control.
- Integrated rEV spiking into bodily fluids prior to EV analysis using fluorescence, nucleic acid, and protein measurements.
Main Results:
- rEVs serve as effective internal controls for quantitative and qualitative EV analysis.
- Spiking rEVs in bodily fluids allows for mapping technical variability and facilitates inter-laboratory studies.
- Demonstrated rEVs' utility in method development, data normalization, and pre-analytical variable assessment.
Conclusions:
- Recombinant EVs (rEV) offer a valuable, standardized biological reference material for extracellular vesicle research.
- This protocol enables improved reliability, reproducibility, and comparability of EV-based studies.
- The rEV approach supports the development and validation of regulated EV applications.
