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작성일 : 17-02-08
[HansaBioMed Life Sciences] PURE-EVs, Size Exclusion Chromatography columns
For isolating & purifying exosomes and extracellular vesicles (EVs)

 

PURE-EVs(Extracellular Vesicles), Size Exclusion Chromatography columns

For isolating & purifying exosomes and extracellular vesicles (EVs)

 

 

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Size Exclusion Chromatography (SEC) is considered one of the best methods for isolating and purifying

exosomes and extracellular vesicles (EVs) from different matrices. In particular, this technique is very

efficient for separating exosomes from the circulating proteins and does not affect the original shape and

functionality of the vesicles.

HBM has developed diff erent classes of SEC columns for EV purifi cation: PURE-EV and miniPURE-EV are

two diff erent size of columns for EV purifi cation from small volume (1.5 ml up to 100 μl) of different matrices.

In addition, HBM provides PURE-EV PLUS, which, together with the SEC column, includes MWCO (molecular

weight cut-off ) concentrators especially designed for concentrating the EV preparation. Using PURE-EVs

PLUS you can get in approximately 15 minutes a concentrate EV preparation.

 

 

Features

• Easy and fast protocol (turnaround time approximately 15 minutes).

• Isolate exosome from small volumes of sample.

• PURE-EVs PLUS includes MWCO concentrators especially designed for EV concentration.

• Reusable up to 5 times.

• Easy to store and ship (4°C).

 

 

Applications

• Exosome isolation from biofl uids and cell media.

• Purifi cation of pre-isolated EVs from contaminants

• Isolated exosomes are suitable for multiple analysis (NTA, ELISA, FACS, WB, EM, MS,

  nucleic acid extraction, etc).

 

 

PURE-EVs, SEC columns performance and applications

 

PURE-EVs, isolation of highly pure exosomes in approximately 15 minutes.

PURE-EVs column was rinsed with 1 ml of human plasma, 24 fractions (500 μl each one) have been

collected and analyzed by ELISA ExoTESTTM assay (see section 3, ExoTEST quantifi cation kit) and by

BCA test for determining respectively the exosome and total protein content. As shown in fi gure 1 and 2, EVs

are eluted in fractions 6 - 11 (turnaround time approximately 15 min), whereas the BCA analysis showed that

the peak of plasma proteins corresponded to the fractions 14 - 24.

The 24 fractions were collected in 4 groups (1-6; 7-12; 13-18; 19-24) and analyzed by NTA with Nanosight.

Figures 3, 4 and 5 show the correlations between the eluted fractions and the performed analysis: positive

matching between ExoTESTTM results and the particles analysis by NTA (fig 3), complete separation of EV

from the plasma circulating proteins (fig 4 and 5).

 

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Fig. 1. Exosome isolation from human plasma by PURE-EVs columns.

 

 

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Fig. 2. Matching of EV quantity and total protein content eluted in each single fraction. ExoTESTTM analysis

shows that EVs are eluted in fractions 6-11 and succesfully separated by the plasma circulating proteins

(eluted in fractions 14-24). ExoTEST results expressed in ratio-to-background.

 

 

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Fig. 3. EV elution peak. ExoTESTTM vs NTA analysis

 

 

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Fig. 4. EV elution vs circulating protein elution. NTA analysis compared to protein BCA test

 

 

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Fig. 5. EV elution vs circulating protein elution. ExoTESTTM analysis compared to protein BCA test

 

 

PURE-EVs PLUS: make your EVs suitable for multiple analyses.

PURE-EVs PLUS inlcludes a MWCO concentrator especially designed for concentrating EVs once eluted by

the SEC column. Concentrator allows concentrated EVs preparation to the level required for diff erent

analyses such as western blotting, nucleic acids extraction, mass spectrometry etc.

 

 

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Fig. 6. NTA analysis of exosome eluted by PURE-EVs column not concentrated and concentrated 10X with

MWCO concentrator.

 

 

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Fig. 7. WB of common exosome markers in 20μl of concentrated EVs.

 

 

 

Ordering informations

 

- PURE-EV, Size Excluson Chromatography columns

Catalog No.

Volume

Size

HBM-PEV-6

1.5 ml ~ 500 μl

6 columns

HBM-PEV-12

12 columns

 

- miniPURE-EV, Size Exclusion Chromatography columns

Catalog No.

Volume

Size

HBM-mPEV-11

500 μl - 100 μl

11 columns

HBM-mPEV-22

22 columns

 

- PURE-EV PLUS, Size Exclusion Chromatography column and MWCO concentrator

Catalog No.

Volume

Size

HBM-PEV-P5

1.5 ml - 500 μl

5 columns + 5 MWCO concentrators

HBM-PEV-P10

10 columns + 10 MWCO concentrators

 

- miniPURE-EV PLUS, Size Exclusion Chromatography column and MWCO concentrator

Catalog No.

Volume

Size

HBM-mPEV-P10

500 μl - 100 μl

10 columns + 10 MWCO concentrators

HBM-mPEV-P20

20 columns + 20 MWCO concentrators

 

 

 

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