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Comparison of advantages and disadvantages of exosome separation method of high-speed centrifuge:
Differential centrifugation: The main steps are as follows: high-speed centrifuge
Density gradient centrifugation
Size exclusion chromatography separation in vitro
Polymer-based precipitation immunosegregation method: screening separation method: high-speed centrifuge Source: Suzhou Alpha Bio.
com
| Separation method | Separation mechanism | Advantages of separation technology | Disadvantages of the separation method |
| Differential centrifugation | The method consists of several centrifugation steps designed to remove cells, large vesicles and debris and precipitate exosomes . |
Differential centrifugation is a standard and very commonly used method for isolating exosomes from biological fluids and media. |
The method is less efficient when analyzed using viscous biological fluids such as plasma and serum. |
| Density gradient centrifugation | This method combines ultracentrifugation with a density gradient of sucrose or iodixanol. |
This method allows the separation of low-density exosomes from other vesicles, particles, and contaminants. |
Very sensitive to centrifugation time. |
| Dimensional exclusion chromatography | Size exclusion chromatography separates macromolecules according to their size . It uses a column filled with porous polymer beads. |
This method allows for the precise separation of large and small molecules and the application of various solutions . In contrast to the centrifugation method, the structure of exosomes separated by chromatography is not affected by shear forces. |
This method requires a long run time, which limits the application of chromatographic separation in the processing of multiple biological samples . |
| filtration | Ultrafiltration membranes are used to separate exosomes from proteins and other macromolecules. Exosome populations are concentrated on membranes . |
Filtration allows the separation of small particles and soluble molecules from exosomes . In this process, exosome populations are concentrated by a filter membrane. |
Exosomes can adhere to the filter membrane and are lost in the following analysis. In addition, exosomes may deform or be damaged due to the additional force applied to allow the analyzed liquid to pass through the membrane. |
| Polymer-based precipitation | The technique includes mixing biological fluids with a polymer-containing precipitate solution, incubation steps, and low-speed centrifugation . |
Advantages of precipitation include mild effects on isolated exosomes and the use of neutral pH. |
Polymer-based precipitation methods work together to isolate non-vesicle contaminants, including lipoproteins . In addition, the presence of polymer materials may not be compatible with downstream analysis. |
| Immunoseptosis | Various immunological methods are applied. Magnetic beads that bind to specific antibodies are used to isolate exosomes . In addition, an ELISA-based separation method has been developed. |
This method allows isolation of all exosomes or selective subtypes of exosomes . In addition, it can be used for the characterization and quantification of exosome proteins. |
This method is not suitable for large sample sizes . In addition, isolated vesicles may lose their functional activity . |
| Screening separation | Screening separation technology is to separate exosomes by membrane screening and filtration by pressure, electrophoresis and other methods . |
Relatively short separation times and provides high purity for exosome isolation . |
Isolated exosome recovery is low . |
Table 1
.
Comparison
of exosome isolation methods.
.
Comparison
of exosome isolation methods.
Differential centrifugation: The main steps are as follows: high-speed centrifuge
Fig.
1.
Differential centrifugation
.
1.
Differential centrifugation
.
Density gradient centrifugation
Figure 2.
Ciliated porous structures for the separation of exosomes
.
This structure does not allow cells and other particles larger than 1 μm to enter the wiring area
.
Some smaller particles and cell debris can enter the micro-column region but are excluded by the nanocilia, forming pores
with a diameter of 30-200 nm.
Cilia structures selectively capture exosomes and small extracellular vesicles
.
Ciliated porous structures for the separation of exosomes
.
This structure does not allow cells and other particles larger than 1 μm to enter the wiring area
.
Some smaller particles and cell debris can enter the micro-column region but are excluded by the nanocilia, forming pores
with a diameter of 30-200 nm.
Cilia structures selectively capture exosomes and small extracellular vesicles
.
Size exclusion chromatography separation in vitro
Figure 3.
Screening model
for exosome filtration.
Screening is done by (a) applying pressure to the on-chip filter or (b) generating an electric field that forces exosomes through the porous membrane, combined with cross-current and electrophoretic sorting
[.
Screening model
for exosome filtration.
Screening is done by (a) applying pressure to the on-chip filter or (b) generating an electric field that forces exosomes through the porous membrane, combined with cross-current and electrophoretic sorting
[.
Polymer-based precipitation immunosegregation method: screening separation method: high-speed centrifuge Source: Suzhou Alpha Bio.
com
