Protein Technologies – Bioseparation Products
Perfluorosorb® S - Unique synthetic adsorbent for the purification of plasmid DNA
Gene therapy and DNA vaccination are rapidly evolving practices as innovation in biotechnology and an industrys dedication to science continues to seek cures for genetic disorders and infectious disease.
ProMetic BioSciences is proud to play a role in the advancement of life science. With the introduction of Perfluorosorb® S, we have applied our core capabilities in the design and manufacture of synthetic, particulate chromatographic media to this important field.
Perfluorosorb® S is a unique, patented product for the purification of plasmid DNA. It has been specifically engineered to preferentially adsorb supercoiled DNA from mixtures of RNA, endotoxin, proteins and other isoforms of DNA and thus provides DNA of the highest purity and potency for laboratory experimentation, clinical trial, and therapeutic manufacture.
Introducing Perfluorosorb® S
Perfluorosorb® S is an engineered, polymeric adsorbent for the purification of plasmid DNA using reverse phase chromatography. Its highly porous structure, broad pH resistance and high binding capacity make it ideal for purification of supercoiled pDNA. Perfluorosorb® S is conveniently available as a slurry in 24% Ethanol in water for improved results in a number of purification processes:
- Primary capture, post-cell lysis and clarification
- As a polishing agent after ion exchange to remove:
- endotoxin
- genomic DNA
- occ and other DNA isoforms
- protein
- RNA
Perfluorosorb® at work
Plasmid DNA binds to Perfluorosorb® in neutral buffers containing ion-pairing agents. Endotoxins are mainly removed in the flow-through and residual amounts are cleared along with RNA in a wash step.
The supercoiled form of plasmid DNA preferentially binds to Perfluorosorb® using reverse phase buffers. Ion-pairing agents are used to enhance separations and resolution in reverse phase chromatography and are totally removed by the wash buffers.
The addition of electrolytes, which are usually present after an ion-exchange step, will actually positively affect the binding between the extremely hydrophobic surface and exposed polar surface of the separation target.
Bioavailability
The transfection efficiency of plasmid DNA purified using Perfluorosorb® S is at least as good as pDNA obtained using other methods. For example, CCL39 fibroblasts that were transiently transfected with a plasmid encoding for Green Fluorescent Protein showed that transfection efficiencies of more 60% could be achieved using plasmids isolated using Perfluorosorb® S.
The proof is in the purity
As an example of the exceptional ability of Perfluorosorb® S to perform at process or laboratory scale, following is a typical purity profile of polished DNA that may be achieved:
- > 95% supercoiled DNA
- 1.8 < A260/280 < 2.0
- < 2% genomic DNA
- < 3 EU endotoxin/mg pDNA
- < 2% protein
- Proper restriction pattern
- Co-migrates with reference
Get started on the road to better results
No special equipment is needed to get started on plasmid purification with Perfluorosorb® S. Simply pack into a standard column using 10µm frits. Follow the supplied packing instructions and special buffer protocols. Perfluorosorb® S columns can be run on any low pressure chromatography workstation. In fact, Perfluorosorb® S is strong enough for use in an HPLC environment, if desired. If you intend to use Perfluorosorb® S in production of cGMP plasmid DNA, ask for ProMetics Drug Master File reference. A Material Safety Data Sheet (MSDS) is also available.
Product Specifications
- New synthetic, fluorinated polymeric entity
- Spherical, porous particles
- Mean particle size: 45µm
- Pure pDNA binding capacity: up to 0.7mg/ml
- Flow rates: up to 700 cm/hr at 1 bar (15psi)
- Stable in pH range 1 - 14
- Resistant to 5M sodium hydroxide
- Suited to repeat-use bioprocess applications
- Store at 2 - 30°C in 24% ethanol
Ordering Information
Perfluorosorb® S (suspension for packing into columns)
| 25 ml | 5003-00025 |
| 100 ml | 5003-00100 |
| 500 ml | 5003-00500 |
| 1000 ml | 5003-01000 |
For more information, please contact us.