Vaccines - Polysaccharides

Primary Clarification

• Remove whole cells and solids from bioreactor fluid and recover product in filtrate
• Requires high capacity
• Wide feed stream variability depending on cell culture type and conditions
• If solid content is too high, centrifugation becomes preferred technology

Recommended Filter

Prostak Open-Channel Modules with Durapore 0.22, 0.45 or 0.65 µm Membrane Filter Characteristics Best suited for large bioreactors >1,000 L or more than 10 batches/year Open-channel design to accommodate high cell density and high solids cell culture fluids High flux at low transmembrane pressure Allows volume reduction as fluid is concentrated Robust construction enables multiple use Validated for multiple SIP cycles Selection of Durapore membrane depends on particle size distribution in feed solution. Choice should be based upon small-scale testing

Secondary Clarification

• Cell recovery from the fermenter broth, elimination of waste fluid in the filtrate
• High solids content in feed stream
• Feed stream quality can vary depending on cell type and culture conditions

Recommended Filter

Triple-layer Millistak+ HC Filters (Grade B1HC)       Disc Filters :: Pod Filters Filter Characteristics Triple-layer configuration includes a membrane bottom layer to enhance filter loading capacity and retention Unique design enables the compression of multiple steps to one, lowering overall filtration costs

Bioburden Reduction

• Reduces bioburden at intermediate process step to reduce the risk of pyrogenic contamination and damage
• Protects downstream filters, media and equipment
• Preserves product purity
• Sterilizing-grade filtration required

Recommended Filter

Millipore Express SHC 0.5 µm + 0.2 µm Filters       Cartridge Filters :: Opticap Capsules :: Optiscale Capsules Filter Characteristics Sterilizing-grade PES 0.2 µm membrane with an additional 0.5 µm prefilter layer all in one pleated device may eliminate the need for a prefilter Designed for bulk intermediate processing where fluids may be fouling in batch sizes ranging from 100 to 20,000 L. 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 pore structure delivers extremely high flow rates at low pressure drop Can be integrity tested on-site

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 Characteristics 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.

Prefiltration (Bulk intermediate)

• Removes colloidal and particulate contaminants and extend the life of final filters
• Should preserve polysaccharide purity and operate consistently at high differential pressures and flow rates
• Must be able to withstand multiple SIP or hot water sanitization cycles
• Run efficiently to ensure process economics

Recommended Filter

Double-layer Milligard Filters (Grade LS, 0.5 µm + 0 µm)       Cartridge Filters :: Opticap Capsule Filters :: Optiscale Capsule Filters Filter Characteristics Prefiltration for Durapore 0.22 µm Filters Double-layer pleated configuration of mixed esters of cellulose membranes provide high dirt-holding capacity, and retention efficiency Low protein-binding media preserves polysaccharide purity High porosity membranes operate at high flow rates and low pressure drops Non-fiber releasing structure preserves product purity 100% integrity tested to ensure quality Robust construction validated to withstand multiple SIP or hot water sanitization cycles Grade LS is commonly used for this application; Milligard filters are available in numerous pore sizes and in single-layer or multi-layer configurations to accommodate the needs of each user, the degree of protection required for the downstream process and the plugging characteristics of the fluid. Choice should be based on small-scale testing.