{"id":18374,"date":"2023-10-17T11:31:36","date_gmt":"2023-10-17T03:31:36","guid":{"rendered":"https:\/\/www.rocker.com.tw\/?post_type=application&p=18374"},"modified":"2026-03-09T11:16:13","modified_gmt":"2026-03-09T03:16:13","slug":"exosome-isolation","status":"publish","type":"application","link":"https:\/\/www.rocker.com.tw\/en\/application\/exosome-isolation\/","title":{"rendered":"Exosome Isolation"},"content":{"rendered":"

Exosomes isolation methods<\/h2>\n

 <\/p>\n

It is critical to understand the importance of exosome isolation methods, which play key roles in cell-to-cell communication and disease mechanisms. Exosomes are tiny vesicles released by cells that contain biomolecules such as proteins and sequences that can affect the functions of surrounding cells. By understanding how exosomes are isolated, we can study the mechanism of cell-to-cell message transmission, thereby revealing the occurrence and development of many diseases.<\/p>\n

The study of exosomes is not only of great significance to basic science, but also expected. Therefore, an in-depth understanding of exosome isolation methods will not only help reveal the mysteries of life, but also has the potential to promote innovation and progress in the medical field.\u00a0<\/p>\n

 <\/p>\n

\n
\n
\n
<\/div>\n<\/div>\n<\/form>\n<\/div>\n
Contents<\/strong><\/span>
\nWhat are exosomes\/extracellular vesicles?<\/a>
\nNomenclature and biological origin of extracellular vesicles<\/a>
\nMethods and differences in isolating exosomes<\/a>
\nConventional ultrafiltration VS tangential flow filtration (TFF)<\/a>
\nApplications of exosomes<\/a><\/div>\n

 <\/p>\n

 <\/p>\n

<\/a>What are exosomes\/extracellular vesicles?<\/h2>\n

 <\/p>\n

It is worth noting that since the biological origin of various vesicles is difficult to clearly identify, the International Society for Extracellular Vesicles (ISEV) recommended in the MISEV2018 guideline that they be named in the following way:<\/p>\n

 <\/p>\n

\n

 <\/p>\n

<\/a>Nomenclature and biological origin of extracellular vesicles<\/h2>\n

 <\/p>\n

It is worth noting that since the biological origin of various vesicles is difficult to clearly identify, the International Society for Extracellular Vesicles (ISEV) recommended in the MISEV2018 guideline that they be named in the following way:<\/p>\n\n\n\n\n
\n

Naming basis<\/p>\n<\/td>\n

\n

Physical characteristics<\/p>\n<\/td>\n

\n

Surface markers; biochemical composition<\/span><\/p>\n<\/td>\n

\n

Descriptions of conditions or cell of origin<\/span><\/p>\n<\/td>\n<\/tr>\n

\n

Example<\/p>\n<\/td>\n

\n

\u2022Exosome Size: small \/ medium \/ large extracellular vesicles (sEVs \/ mEVs \/ lEVs)<\/p>\n

\u2022Density: slow \/ middle, high EVs<\/p>\n<\/td>\n

\n

\u2022D63+ \/ CD81+ EVs<\/p>\n

\u2022Annexin A5-stained EVs<\/p>\n<\/td>\n

\n

\u2022 Podocyte EVs<\/p>\n

\u2022 Hypoxic EVs<\/p>\n

\u2022 Large oncosomes<\/p>\n

\u2022 Apoptotic bodies<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\"the<\/p>\n

 <\/p>\n

What is the difference between liposomes and EVs?<\/h3>\n

 <\/p>\n

Compared with synthetic carriers such as liposomes and nanoparticles, exosomes have characteristics such as endogeneity and heterogeneity, making exosomes useful as Excellent carriers can transport bioactive substances to target cells through a variety of pathways and sites to participate in regulation, such as tissue repair, Immune regulation, angiogenesis, cellular differentiation, neoplasia, etc.<\/p>\n

 <\/p>\n

Advantages and disadvantages of exosomes<\/h3>\n

 <\/p>\n

Therefore, exosomes have great potential and advantages in disease diagnosis, treatment and biology research, such as as biomarkers for tumor diagnosis and drugs for cancer treatment. carrier etc. However, exosomes also have some limitations, such as low stability, low yield, low purity, and weak targeting, which may limit their clinical application.<\/p>\n\n\n\n\n\n
\n

Extracellular vesicles, EVs<\/p>\n<\/td>\n

\n

Exosomes<\/p>\n<\/td>\n

\n

Microvesicles<\/p>\n<\/td>\n

\n

Apoptotic bodies<\/p>\n<\/td>\n<\/tr>\n

\n

Size<\/p>\n<\/td>\n

\n

40~120 nm<\/p>\n<\/td>\n

\n

100~1000 nm<\/p>\n<\/td>\n

\n

1000~5000 nm<\/p>\n<\/td>\n<\/tr>\n

\n

Biogenesis<\/p>\n<\/td>\n

\n

Multivesicular bodies, MVB
\nEndo-lysosomal pathway<\/p>\n<\/td>\n

\n

Plasma membrane, outward budding<\/p>\n<\/td>\n

\n

Plasma membrane, apoptosis process<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

\n

 <\/p>\n

<\/a>Exosomes isolation methods<\/h2>\n

 <\/p>\n

As the research on exosomes isolation continues to deepen, their potential application value is also continuously being explored. The isolation, purification, and concentration (also known as enrichment) of exosomes are critical for evaluating their biological functions and their downstream applications. However, the components of biological samples are complex and contain many similar molecular structures, such as cell fragments, protein aggregates, lipoproteins, etc., and the size and composition of exosomes are heterogeneous, making “isolation” more challenging. Therefore, “how to effectively isolate and concentrate exosomes” is a major test faced in academic research and clinical applications.<\/p>\n

 <\/p>\n

Differences between exosomes isolation methods<\/h3>\n

 <\/p>\n

There are many exosomes isolation methods, depending on the principles. Common exosomes isolation methods are ultracentrifugation, particle size screening chromatography, ultrafiltration, precipitation and immunoaffinity. The following table summarizes the differences between the methods:<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

 <\/p>\n<\/td>\n

\n

Ultracentrifugation
\n(UC) *1<\/sup><\/p>\n<\/td>\n

\n

Particle size screening chromatography (SEC)<\/p>\n<\/td>\n

\n

Ultrafiltration (UF)<\/p>\n<\/td>\n

\n

Precipitation<\/p>\n<\/td>\n

\n

Immunoaffinity (IA)<\/p>\n<\/td>\n<\/tr>\n

\n

Principle<\/p>\n<\/td>\n

\n

Sedimentation Coefficient (size, density)<\/p>\n<\/td>\n

\n

Hydrodynamic Radius (size, molecular weight)<\/p>\n<\/td>\n

\n

Membrane pore size (size, molecular weight)<\/p>\n<\/td>\n

\n

Solubility (surface charge)<\/p>\n<\/td>\n

\n

Specific Binding (Membrane protein marker)<\/p>\n<\/td>\n<\/tr>\n

\n

Yield<\/p>\n<\/td>\n

\n

Low<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n

\n

High<\/p>\n<\/td>\n

\n

High<\/p>\n<\/td>\n

\n

Low<\/p>\n<\/td>\n<\/tr>\n

\n

Purity<\/p>\n<\/td>\n

\n

Medium (lipoprotein)<\/p>\n<\/td>\n

\n

Medium to high (lipoprotein, albumin)<\/p>\n<\/td>\n

\n

Medium (protein)<\/p>\n<\/td>\n

\n

Low (proteins, polymers)<\/p>\n<\/td>\n

\n

High<\/p>\n<\/td>\n<\/tr>\n

\n

Functionality<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n

\n

High<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n

\n

Low<\/p>\n<\/td>\n

\n

Low<\/p>\n<\/td>\n<\/tr>\n

\n

Time<\/p>\n<\/td>\n

\n

> 4 hr<\/p>\n<\/td>\n

\n

0.3 hr*2<\/sup><\/p>\n<\/td>\n

\n

< 4 hr<\/p>\n<\/td>\n

\n

\u22480.3~12 hr<\/p>\n<\/td>\n

\n

4~20 hr<\/p>\n<\/td>\n<\/tr>\n

\n

Sample volume<\/p>\n<\/td>\n

\n

Large<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n

\n

Large *(TFF)<\/sup><\/p>\n<\/td>\n

\n

Large<\/p>\n<\/td>\n

\n

Small<\/p>\n<\/td>\n<\/tr>\n

\n

Denature and inactive of EVs<\/p>\n<\/td>\n

\n

Yes (high speed)<\/p>\n<\/td>\n

\n

No<\/p>\n<\/td>\n

\n

Yes (Shear force)<\/p>\n<\/td>\n

\n

Yes (Co-precipitates with proteins or polymers)<\/p>\n<\/td>\n

\n

Yes (Refining steps)<\/p>\n<\/td>\n<\/tr>\n

\n

Complexity<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n

\n

Simple<\/p>\n<\/td>\n

\n

Simple<\/p>\n<\/td>\n

\n

Simple<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n<\/tr>\n

\n

Scalability<\/p>\n<\/td>\n

\n

Medium<\/p>\n<\/td>\n

\n

High<\/p>\n<\/td>\n

\n

Medium to high 3(TFF)<\/sup><\/p>\n<\/td>\n

\n

High<\/p>\n<\/td>\n

\n

Low<\/p>\n<\/td>\n<\/tr>\n

\n

Other<\/p>\n<\/td>\n

\n

\u2022 Labor intensive and time consuming<\/p>\n<\/td>\n

\n

\u2022 High reproducibility<\/p>\n

\u2022 EVs have complete functions and forms<\/p>\n<\/td>\n

\n

\u2022 High reproducibility<\/p>\n

\u2022 High flexibility of use<\/p>\n

\u2022 Filter membrane clogged<\/p>\n<\/td>\n

\n

\u2022 Polymers are difficult to separate<\/p>\n

\u2022 More costly kit<\/p>\n<\/td>\n

\n

\u2022 EVs have the best purity<\/p>\n

\u2022 Study of specific EV subpopulations<\/p>\n

\u2022 Unable to separate total EVs<\/p>\n

\u2022 Antibodies are expensive<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

*1 Ultracentrifugation (UC) is also called differential centrifugation (dUC)
\n*2 EVs purification can be completed within 20 minutes using commercially available kits
\n*3 High scalability refers to TFF tangential flow filtration method<\/p>\n

 <\/p>\n

\n

 <\/p>\n

 <\/p>\n

<\/a>Conventional ultrafiltration VS tangential flow filtration<\/h2>\n

 <\/p>\n

Ultrafiltration (UF) is a method of separation based on molecular size, which can be subdivided into centrifugal-driven ultrafiltration centrifuge tubes, pressure-driven stirred filters, and tangential flow filtration (TFF). type. Ultrafiltration centrifugal tubes and stirred filters are easy to operate and have fast processing time, but the biggest problem is that the filter membrane is easily clogged, which not only reduces the separation efficiency but also increases the cost; and the development of tangential flow filtration technology (TFF) It can significantly solve the problem of filter membrane clogging.<\/p>\n

\"exosomes<\/p>\n

 <\/p>\n

Using tangential flow filtration to separate EVs<\/h3>\n

 <\/p>\n

Many documents also point out that when TFF is used to separate EVs, its yield, reproducibility, and decontamination capabilities are superior to the ultracentrifugation method (UC or dUC) called the gold standard for EV purification, and even saves more than 40% of time. , more suitable for large-scale mass production applications.
\nFor a detailed comparison of each method and the EV isolation data of TFF & UC, you can watch the “Exosome Exosome Purification Online Seminar Live Highlights”.<\/p>\n

 <\/p>\n

\n\n\n\n\n
\n

ROCKER Tanfil 100 Tangential Flow Filtration System
\n\u25c6 Plug & start all-in-one system
\n\u25c6 Concentration and diafiltration at same run.
\n\u25c6 Suitable for bioprocesses including DNA and RNA concentration
\n\u25c6 Easy scale-up. Use same parameters for mass production.
\n\u25c6 No more membrane fouling, longer lifespan.
\n\u25c6 CE, RoHS, UKCA certificated<\/p>\n<\/td>\n

\n

\"lab<\/p>\n<\/td>\n<\/tr>\n

Lab scale tangential flow filtration (TFF) system – Tanfil 100<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/blockquote>\n

\"try<\/a><\/p>\n

 <\/p>\n

\"Rocker<\/a>
Global expert support across USA, Asia, and worldwide. \u25b2Click to get a quick quote from your local Rocker dealer.<\/figcaption><\/figure>\n

 <\/p>\n

Improve the exosomes isolation efficiency<\/h2>\n

 <\/p>\n

Although various isolation methods have been developed, these methods still have shortcomings and cannot fully meet the high purity and high yield requirements for isolating exosomes. In order to improve the isolation efficiency and concentration of exosomes, many research teams have begun to try to combine multiple isolation methods; so far, studies have pointed out that multiple isolation methods can increase yield and purity more effectively than a single isolation method.<\/p>\n

 <\/p>\n

Exosome Isolation via TFF Using Diverse Technologies<\/h3>\n

 <\/p>\n

For example, UC and UF are combined to initially extract exosomes, and then IA is used to further purify target exosomes. Alternatively, UC and SEC can be combined to not only process large sample volumes but also effectively reduce interference from contaminants. When selecting a combination method, attention should be paid to whether each single method will cause adverse effects on the target exosomes, such as exosome denaturation and inactivation. The following are\u00a0exosomes isolation methods using TFF with various technologies.<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\u00a0<\/td>\n\n

Target<\/p>\n<\/td>\n

\n

EV approach<\/p>\n<\/td>\n

\n

TFF function<\/p>\n<\/td>\n

\n

Pore size<\/p>\n<\/td>\n

\n

Brand name<\/p>\n<\/td>\n

\n

Sample source<\/p>\n<\/td>\n<\/tr>\n

\n

Purification<\/p>\n<\/td>\n

\n

Concentration<\/p>\n<\/td>\n

\n

Buffer<\/p>\n<\/td>\n<\/tr>\n

\n

1<\/p>\n<\/td>\n

\n

Exosome<\/p>\n<\/td>\n

\n

TFF \u2192 SEC<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

5 nm<\/p>\n<\/td>\n

\n

HansaBioMed,<\/p>\n

TFF-Easy<\/p>\n<\/td>\n

\n

Cell culture medium (DP-MSC)<\/p>\n<\/td>\n<\/tr>\n

\n

2<\/p>\n<\/td>\n

\n

EVs<\/p>\n<\/td>\n

\n

TFF \u2192 SEC<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

50 \u00b1 10 nm<\/p>\n<\/td>\n

\n

HansaBioMed,<\/p>\n

TFF-EVs<\/p>\n<\/td>\n

\n

HPL Supernatant<\/p>\n<\/td>\n<\/tr>\n

\n

3<\/p>\n<\/td>\n

\n

Exosome<\/p>\n<\/td>\n

\n

UC \u2192 TFF<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

V<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

5 nm<\/p>\n<\/td>\n

\n

HansaBioMed,<\/p>\n

TFF-Easy<\/p>\n<\/td>\n

\n

Cell culture medium (NK-92 cell)<\/p>\n<\/td>\n<\/tr>\n

\n

4<\/p>\n<\/td>\n

\n

Plant EVs<\/p>\n<\/td>\n

\n

dUC \u2192 TFF<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

V<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

5 nm<\/p>\n<\/td>\n

\n

HansaBioMed,<\/p>\n

TFF-Easy<\/p>\n<\/td>\n

\n

Cell culture medium (tobacco)<\/p>\n<\/td>\n<\/tr>\n

\n

5<\/p>\n<\/td>\n

\n

Exosome<\/p>\n<\/td>\n

\n

(TFF) \u2192 Qiagen, exoEasy Maxi Kit \/ SEC<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

V<\/p>\n

(10~20X)<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

5 nm<\/p>\n<\/td>\n

\n

HansaBioMed,<\/p>\n

TFF-Easy<\/p>\n<\/td>\n

\n

Cell culture medium (cancer cells)<\/p>\n<\/td>\n<\/tr>\n

\n

6<\/p>\n<\/td>\n

\n

Exosome<\/p>\n<\/td>\n

\n

LSC* (300g) \u2192 TFF\u2573<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

100 kDa<\/p>\n<\/td>\n

\n

Pall, Minimate<\/p>\n<\/td>\n

\n

Cell culture medium (EF-MSCs)<\/p>\n<\/td>\n<\/tr>\n

\n

7<\/p>\n<\/td>\n

\n

EVs<\/p>\n<\/td>\n

\n

LSC* (300g) \u2192 TFF<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\n

V
\n(50X)<\/p>\n<\/td>\n

\u00a0<\/td>\n\n

300 kDa<\/p>\n<\/td>\n

\n

Pall, Minimate<\/p>\n<\/td>\n

\n

Cell culture medium\u00a0(ASC)<\/p>\n<\/td>\n<\/tr>\n

\n

8<\/p>\n<\/td>\n

\n

EVs<\/p>\n<\/td>\n

\n

TFF<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\n

V
\n(4~20X)<\/p>\n<\/td>\n

\n

V<\/p>\n<\/td>\n

\n

0.65 \u03bcm, 500 kDa<\/p>\n<\/td>\n

\n

Spectrum Labs<\/p>\n<\/td>\n

\n

Cell culture medium<\/p>\n

Lipoaspirate<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*LSC\uff1aLow speed centrifugation<\/p>\n\n\n\n\n
\n

1 Klimova, D. et al., 2023<\/a><\/p>\n<\/td>\n

\n

2 Kenneth W. Witwer, 2022<\/a><\/p>\n<\/td>\n

\n

3 Kim et al., 2022<\/a><\/p>\n<\/td>\n

\n

4 Eric With, et al., 2021<\/a><\/p>\n<\/td>\n<\/tr>\n

\n

5 Altanerova, U. et al., 2021<\/a><\/p>\n<\/td>\n

\n

6 Sung et al., 2021<\/a><\/p>\n<\/td>\n

\n

7 Lee et al., 2021<\/a><\/p>\n<\/td>\n

\n

8 Busatto, S. et al., 2018<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n