Developing cell lines that produce commercially acceptable yields can be difficult and labor-intensive, so the selection of a high-producer cell line is critical. ?

Vaccine manufacturers rely on proper high-throughput development tools to confidently select a stable and safe cell line that meets regulatory requirements.

Selection of the appropriate raw materials is essential to ensure process safety during cell-line generation, and many in industry are moving toward chemically defined (CD) media to support process safety and robustness.

Adequate and compliant cell banking and testing is an industry imperative to ensure sufficient quality supply for as long as the product is needed.

The flexibility and functionality of Ambr^? 15 & Ambr^? 250 can accelerate cell line development and process optimization. Ambr^? is the only high-throughput cell culture system with scalable, multi parallel mini-bioreactors and automated functionality.

• Proven technology with all suspension cell lines and for fermentation applications
• Capable in all operation modes: batch, fed-batch, perfusion
• Integrated with a wide range of connectivity of inline analytics, including BioPAT^? Spectro ?
• Available with MODDE^? software to facilitate DoE ?

Additional solutions to consider: ?

• With more than 70 experienced scientists in the US and Europe to conduct a full package of cell line and product testing compliant with current regulations, Sartorius is well-positioned to support customers, from cell line development and cell banking to commercial manufacturing.
• Sartorius’ cell line development services ensure the best clone selection for high-productivity stable cell lines and protein expression in CHO DG44 when CHO cell lines are used.
• Sartorius offers a new generation of chemically defined (CD) and animal component-free (ACF) cell culture media for mammalian and insect cell lines developed and tested with Sartorius bioreactors.

Seed expansion from cell bank to production bioreactor is critical to effectively seed the production bioreactor. The latest innovations use process intensification to reduce the seed train and ensure the highest possible process consistency while Process Analytical Technologies enable full process control.

Meanwhile, robust & high yield protein production relies on scalability from process development to production scale manufacturing. The use of Process Analytical Technologies enables process controls and monitoring, while implementation of process intensification leads to a higher protein concentration.

In both cases, manufacturers need to:

• Ensure high productivity to allow production out of smaller bioreactors and decrease overall costs
• Ensure full scalability from process to development to manufacturing scale and demonstrate process robustness
• Demonstrate overall process safety and quality, in compliance to the requirements of the industry
• Ensure flexibility to adapt the production to the demand, especially when facing an outbreak

Biostat STR^? is a single-use bioreactor offering the widest range of PAT in a Biosafety Level 2 environment, making it the perfect match for viral production:

• A unique and robust film across the process ensures excellent cell growth
• Demonstrated for mammalian cell lines, insect cell lines, as well as fermentation applications with low demand in oxygen ?
• Capable in all operation modes: batch, fed-batch and perfusion, allowing process intensification
• Integration of a wide range of PAT, including BioPAT^? Viamass, BioPAT^? Trace and BioPAT^? Spectro
• Built on Biosafety principles to reduce risk of contamination and loss of product ?
• Direct scalability from Ambr^? to STR for rapid process development

Additional USP solutions to consider:

• Biostat^? RM, a fully GMP-compliant, wave-mixed bioreactor, is an easy-to-use system for seed expansion offering the widest range of PAT in a Biosafety Level 2 environment
• Biostat^? D-DCU is a compact bioprocess system available in microbial or cell culture versions with vessel choices from 10 to 200L working volume
• A new generation of chemically defined (CD) and animal component-free (ACF) cell culture media for mammalian and insect cells developed and tested with Sartorius bioreactors

Vaccine developers must develop a downstream process tailored to their antigen, whether it is a protein or a virus-like particle. The choice of expression system will influence the choice of technologies. ?
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In either case, the steps will be the following:

• Harvest and clarification to efficiently remove cell debris, large aggregates, and insoluble contaminants
• Capture and polishing achieve the highest purity and recovery of the antigen
• Crossflow filtration to concentrate the product and exchange buffers before/during chromatography steps or for final formulation
• Sterile filtration to prevent microbial contamination and ensure patient safety

Which leads to the following needs:

• In the absence of an affinity ligand, a wide range of chromatography resins and ligand is needed to find the best purification strategy and ensure a rapid and efficient purification of the antigen ?
• A solution to address the variety of contaminants that will have to be removed, depending on the expression system: DNA, host cell proteins, viruses, baculoviruses, endotoxins, etc.
• High-capacity, high-recovery technologies are needed to optimize overall yield and manage costs, which becomes increasingly challenging as the process is intensified ?
• Ensure flexibility to adapt the production to the demand, especially when facing potential outbreaks

Hydrosart^?, a state-of-the-art crossflow membrane designed for low adsorption of proteins, has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for protein-based vaccines:

• Available with molecular weight cut-off membrane ranging from 2kD up to 300kD ?
• Provides low adsorption, high flow rate and high product recovery
• Compatible with a wide range of chemicals for a broader spectrum of operating conditions and cleaning protocols
• Can be delivered gamma sterile, allowing closed, ready-to-use and sterile processing compliant to Biosafety Level 2
• Available in standard and single-pass format for continuous ultrafiltration and diafiltration

Additional DSP solutions to consider:

• A wide range of chromatography resins for protein capture and the widest range of membranes for VLP capture and polishing
• A unique orthogonal platform for state-of-the-art virus clearance when antigens are produced in mammalian and insect cells
• Virus Counter^? to monitor baculovirus titer for proteins and VLP produced in insect cells
• Sartopore^? Platinum for final filling applications. The unique surface modification of the PES membrane provides the lowest adsorption for high product yield and secure wetting for reliable integrity testing