Related Experiment Video
Updated: Jun 7, 2026
Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry
Published on: March 17, 2015
Detection and isolation of cell-derived microparticles are compromised by protein complexes resulting from shared
Bence György1, Károly Módos, Eva Pállinger
1Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary.
Abstract:
Numerous diseases, recently reported to associate with elevated microvesicle/microparticle (MP) counts, have also long been known to be characterized by accelerated immune complex (IC) formation. The goal of this study was to investigate the potential overlap between parameters of protein complexes (eg, ICs or avidin-biotin complexes) and MPs, which might perturb detection and/or isolation of MPs. In this work, after comprehensive characterization of MPs by electron microscopy, atomic force microscopy, dynamic light-scattering analysis, and flow cytometry, for the first time, we drive attention to the fact that protein complexes, especially insoluble ICs, overlap in biophysical properties (size, light scattering, and sedimentation) with MPs. This, in turn, affects MP quantification by flow cytometry and purification by differential centrifugation, especially in diseases in which IC formation is common, including not only autoimmune diseases, but also hematologic disorders, infections, and cancer. These data may necessitate reevaluation of certain published data on patient-derived MPs and contribute to correct the clinical laboratory assessment of the presence and biologic functions of MPs in health and disease.
Insights
Immune complexes (ICs) share physical properties with microvesicles/microparticles (MPs), potentially interfering with MP detection and quantification in various diseases. This finding necessitates reevaluating existing patient data on MPs.
Area of Science:
- Biomedical Sciences
- Immunology
- Cell Biology
Background:
- Elevated microvesicle/microparticle (MP) counts are linked to numerous diseases.
- Accelerated immune complex (IC) formation is a known characteristic of these diseases.
- Potential overlap between MPs and protein complexes like ICs could affect MP analysis.
Purpose of the Study:
- To investigate the overlap in biophysical properties between protein complexes (ICs) and MPs.
- To assess the impact of this overlap on MP detection, isolation, and quantification.
Main Methods:
- Comprehensive characterization of MPs using electron microscopy, atomic force microscopy, dynamic light-scattering, and flow cytometry.
- Analysis of biophysical properties including size, light scattering, and sedimentation.
Main Results:
- Protein complexes, particularly insoluble ICs, exhibit overlapping biophysical properties with MPs.
- This overlap significantly affects MP quantification by flow cytometry and purification by differential centrifugation.
- The interference is pronounced in diseases with common IC formation, such as autoimmune diseases, infections, hematologic disorders, and cancer.
Conclusions:
- Insoluble ICs share biophysical characteristics with MPs, impacting their accurate measurement.
- Published data on patient-derived MPs may require reevaluation due to potential IC interference.
- Findings may lead to improved clinical laboratory assessment of MPs in health and disease.
