Ultrafiltration and Diafiltration (UF/DF)
Purify & concentrate biomolecules, exchange buffers, and more
Get everything ready for the next step
Ultrafiltration (UF) and diafiltration (DF) are critical for the development and manufacturing of biological therapeutics, such as proteins, antibodies, and nucleic acids as well as therapies that rely on viral or lipid nanoparticle (LNP) delivery. They can be used separately or in combination to get the target molecule ready for the next processing step by enabling sample cleanup, purification, concentration, buffer exchange, and desalting. While both involve the use of membrane filters, they operate in slightly different ways.
What’s the difference between ultrafiltration and diafiltration?
UF uses a semipermeable membrane to separate molecules based on size and is typically used for purification and concentration. DF is the process of adding back a different buffer or solvent after UF.
Here’s how they work:
Use the force
UF uses a semi-permeable membrane with active force applied to drive filtration. Typically, force is applied by centrifugation or pressure.
Size matters
The semi-permeable membrane retains larger target molecules while allowing smaller molecules to filter out. Regenerated cellulose and polyethersulfone (PES) membranes are popular for gene therapy and biologics applications given their hydrophilic properties popular for gene therapy and biologics applications given its hydrophilic properties, low protein binding, and compatibility across a wide pH range.
Exchange that buffer
DF enables buffer exchange by adding new buffer to the retentate. The buffer is replaced, and the sample is returned to the original volume. At this point the UF/DF cycle can be repeated until the sample reaches a targeted level of replacement with the new buffer.
Lil' Tuna
Lil’ Tuna is the first automated, single sample or plate-based ultrafiltration and diafiltration (UF/DF) system for buffer exchanging, concentrating, or desalting proteins, viral vectors and nucleic acids - with consistently high recoveries. Process 1 to 96 samples in parallel with minimal hands-on time and achieve consistent and reliable sample prep for formulation screening, stability testing or any other downstream assay.
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Gene therapy and biologics scientists can now use the right tools for buffer exchange and sample concentration with platforms that free you up and deliver superior results. Have a question or ready to find out more?