Q: What reagents, instruments, and equipment are required for the EV purification using this column?

A: The reagents, instruments, and equipment required for the use of this product are listed below.

Reagents

• MassivEV Purification Buffer Set (Catalog ID: 295-96601)
• Cell culture medium
• EV production medium (Product Number: 053-09451 / 298-84001)
• 100％ Ethanol
• Ultrapure water

Instruments and Equipment-
*The following instruments may or may not be required depending on the operation. For more details, please see instruction manual.

• Centrifuge
• Cell strainer (e.g. FALCON, Product Number: 352340）
• 5 μm filter (e.g. GVS, Product Number: 1215396）
• 0.8 μm filter (e.g. GVS, Product Number: 1214568）
• 0.22 μm filter (e.g. Corning, Product Number: 431118）
• Incubation or water bath (which can be warmed at 37℃)
• Peristaltic pump (e.g. Repligen, Product Number: ACJR-U10-R)
• Fluid transfer tubing (see support document)
• Collection tube (e.g. Corning, Product Number: 430791)
• Ultrafiltration membrane [100kDa/PES material] (e.g. Sartorius, Product Number: VS0141)
• Gel-filtration columns（e.g. Cytiva, Product Number: 28918004）

Q: Do you have any recommendations for tubing suitable for liquid transfer? How should this tubing be connected?

A: We don't have specific recommendations, provided the tubing can control the flow rate and withstand pressure. Ensure the tubing is compatible with your peristaltic pump.

For guidance on connecting the tubing, please refer the support document titled:
Example of a purification system using the MassivEV EV Purification Column PS

Q: How should I choose between centrifugation and filtration for the pretreatment of cell culture supernatants?

A: Your choice between centrifugation and filtration will depend on the facilities, instruments, and equipment your laboratory already has. The decision does not need to be influenced by the sample type or its specific application.

Q: For buffer exchange, how should I choose between gel filtration and ultrafiltration?

A: We recommend gel filtration when the sample is for administration to animals.

Q: What volume of the EV solution is obtained during elution?

A: The volume is 4 mL for a 1 mL column, and 20 mL for a 5 mL column.

Q: How long can a column wrapped in Parafilm be stored?

A: Based on our experience at Fujifilm Wako, a column can be stored for approximately 1.5 months.

Q: Can the column be used multiple times?

A: The column can be reused for the same sample up to 5 times, including the initial use and four subsequent reuses.
For each reuse, begin with Cleaning 1, as detailed in the "Procedures" section (5) of the instruction manual.

Q: How should air that has entered the column be removed?

A: To remove air from the column, try passing 20% or 70% ethanol through it at the flow rates specified below. Note that prolonged or repeated exposure to air can degrade the resin's performance. For more information, consult the instruction manual.

Flow Rates

• 1 mL column: 0.85 mL/min
• 5 mL column: 4.25 mL/min

Q: Can I use this product for commercial or for-profit purposes?

A: Please use this product for research purposes only. If you wish to use it for commercial or for-profit purposes, please contact Fujifilm Wako at wkuslabchem@fujifilm.com.