Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Subcellular Fractionation01:32

Subcellular Fractionation

7.1K
The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...
7.1K
Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

5.8K
Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
5.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Engineering zirconium-based MOFs for selective adsorption of estrogenic contaminants and the application in electrospun solid-phase microextraction.

Journal of hazardous materials·2026
Same author

Protein-Based High-Performance Liquid Chromatography and Cyclodextrin-Capillary Electrokinetic Chromatography for the Chiral Separation of Azoles.

Electrophoresis·2026
Same author

Qualitative gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry fingerprinting of tissue-specific phytochemicals in finnish Quercus robur L. acorns.

Scientific reports·2026
Same author

Identification of adducts formed between phosphatidylcholine and mustard agents.

Analytical and bioanalytical chemistry·2026
Same author

Sample Preparation and Separation of Lignans by Liquid Chromatography.

Journal of separation science·2026
Same author

Metabolic Glycoengineering Enables Fluorine-18 Radiolabeling of T Lymphocytes via Dual-Bioorthogonal Chemistry.

Bioconjugate chemistry·2026

Related Experiment Video

Updated: Jul 31, 2025

Author Spotlight: Asymmetric Field Flow Fractionation for Bioreactor Integration
06:28

Author Spotlight: Asymmetric Field Flow Fractionation for Bioreactor Integration

Published on: February 2, 2024

805

Automated On-Line Isolation and Fractionation Method for Subpopulations of Extracellular Vesicles.

Susanne K Wiedmer1, Evgen Multia1, Thanaporn Liangsupree1

  • 1Department of Chemistry, University of Helsinki, Helsinki, Finland.

Methods in Molecular Biology (Clifton, N.J.)
|May 4, 2023
PubMed
Summary

This study presents an automated method combining immunoaffinity chromatography (IAC) and asymmetrical flow field-flow fractionation (AsFlFFF) for isolating extracellular vesicles from human plasma. The technique achieves high purity and yield of vesicle subpopulations without lipoproteins.

Keywords:
antibodiesasymmetrical flow field-flow fractionationextracellular vesiclesimmunoaffinity chromatographyisolationon-line couplingtetraspanins

More Related Videos

Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles
10:16

Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles

Published on: January 20, 2023

3.1K
Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
10:46

Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media

Published on: May 13, 2022

3.9K

Related Experiment Videos

Last Updated: Jul 31, 2025

Author Spotlight: Asymmetric Field Flow Fractionation for Bioreactor Integration
06:28

Author Spotlight: Asymmetric Field Flow Fractionation for Bioreactor Integration

Published on: February 2, 2024

805
Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles
10:16

Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles

Published on: January 20, 2023

3.1K
Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
10:46

Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media

Published on: May 13, 2022

3.9K

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Immunoaffinity chromatography (IAC) and asymmetrical flow field-flow fractionation (AsFlFFF) are powerful techniques for biomacromolecule isolation and fractionation.
  • Extracellular vesicles (EVs) are crucial biomarkers, but their isolation from complex matrices like human plasma, especially free of lipoproteins, remains challenging.
  • Existing methods may lack the specificity or throughput required for comprehensive EV analysis.

Purpose of the Study:

  • To develop and validate an automated, on-line coupled IAC-AsFlFFF method for the isolation and fractionation of extracellular vesicles from human plasma.
  • To achieve high purity and yield of EV subpopulations, specifically excluding lipoproteins.
  • To establish a fast, reliable, and reproducible platform for challenging biomacromolecule separation.

Main Methods:

  • On-line coupling of immunoaffinity chromatography (IAC) with polymeric monolithic disk columns to selective antibodies.
  • Integration with asymmetrical flow field-flow fractionation (AsFlFFF or AF4) for subsequent fractionation of isolated biomacromolecules.
  • Application to human plasma samples for the isolation and fractionation of extracellular vesicles, ensuring lipoprotein removal.

Main Results:

  • Successful isolation and fractionation of extracellular vesicle subpopulations from human plasma.
  • Demonstrated removal of lipoproteins from the isolated EV fractions, enhancing purity.
  • Achieved high purity and high yields of specific EV subpopulations through the automated, on-line coupled system.

Conclusions:

  • The developed on-line IAC-AsFlFFF methodology provides an efficient and automated approach for isolating and fractionating extracellular vesicles from human plasma.
  • This technique enables the separation of EV subpopulations with high purity and yield, free from interfering lipoproteins.
  • The method offers a reliable and reproducible platform for advancing research in EV biology and diagnostics.