Vaccines - Plasmid DNA

Clarification and Solids Removal (in place of bag filter)

• Remove solids after the cell lysis step
• Have high debris holding capacity
• Protect downstream membranes
• Charged medias should be avoided due to plasmid interaction
• Typically operated at low delta P to minimize precipitate or breakthrough
• Process is more efficient in cases of flocculants that float
• Washing of precipitate to reduce endotoxin and genomic DNA contaminants

Recommended Filter

Polygard CR 10 µm Filters         Cartridge Filters :: Opticap Capsule Filters Filter Requirements For use in place of a bag filter or for extra clarification after cell lysis Multi-stage graded density design retains solids or sediment within the depth matrix Polypropylene media provides excellent solids capacity and high throughput.

Clarigard 0.5 µm Filters         Cartridge Filters :: Opticap Capsule Filters Filter Requirements For use in series after the Polygard CR filter when no bag filter is used for solids removal Polypropylene graded density prefilter for robust particle and colloid removal 1–2 log bioburden reduction Validated for multiple sanitization and SIP cycles Higher filtrate quality allows smaller size requirements for downstream sterilizing-grade filtration step Non-fiber releasing structure preserves product purity.

Concentration and Diafiltration (prior to column chromatography steps)

• Concentration of process fluids and buffer exchange
• Will extend the life of expensive chromatography media and improve their speed and efficiency
• Fast, robust process able to accommodate feed fluid variations
• Wide chemical compatibility with cleaning agents
• Easily scalable from lab to manufacturing scale

Recommended Filter

Pellicon V Screen Cassettes with Ultracel PLC 30 or 100 kD Membrane Filter Requirements Low feed flow requirement minimizes pumping, working and hold up volumes Highly retentive regenerated cellulose membranes offer good permeate flux and withstand process upsets Robust low binding void-free Ultracel PLC membranes assure consistent processing from batch to batch, have good chemical compatibility and are easy to clean Complete range of devices for easy scale-up Choose a membrane molecular weight cut-off 3–5X smaller than the biomolecule. Choice should be based on small-scale testing.

Concentration and Diafiltration (for final formulation)

• Concentration and diafiltration adjusts the end product to the proper dosage conditions prior to final packaging
• Fast, robust process able to accommodate feed fluid variations
• Easily scalable from lab to manufacturing scale

Recommended Filter

Pellicon C Screen Cassettes with Ultracel PLC 30 or 100 kD Membranes Filter Requirements Low feed flow requirement minimizes pumping, working and hold up volumes Higher product recovery and flux Highly retentive regenerated cellulose membranes offer good permeate flux and withstand process upsets Robust low binding void-free Ultracel PLC membranes assure consistent processing from batch to batch, have good chemical compatibility and are easy to clean Complete range of devices for easy scale-up Choose a membrane molecular weight cut-off 3–5X smaller than the biomolecule. Choice should be based on small-scale testing.

Sterile Filtration

• Sterilizing-grade step removes bacteria prior to aseptic fill
• Ensures product safety
• Highly validated and reliable technology
• Preserves product purity

Note: Membrane chemistries other than PES are known to cause gel formation at the membrane surface. We strongly recommend a PES filter for this step.

Recommended Filter

Double-layer Millipore Express SHC 0.5 µm + 0.2 µm Filters        Cartridge Filters :: Opticap Capsules :: Optiscale Capsules Filter Requirements Sterilizing-grade PES 0.2 µm membrane with an additional 0.5 µm PES prefilter layer all in one pleated device Double-layer construction provides excellent capacity even in high salt conditions PES membrane provides broad chemical compatibility across a wide pH range (1–14) Robust construction validated for multiple SIP cycles 100% integrity tested to ensure quality Asymmetric PES pore structure delivers extremely high flow rates at low pressure drop Can be integrity tested on-site