Intensification Strategies Start With a Focus on Process Development
Optimized Scalable Process for Shorter Time to Market
Efficient process development (PD) including a high-titer cell line, clone selection and optimization, and process characterization is critical to develop a scalable upstream process. Now, with technology scalable to fit different intensified process types, PD also enables opportunities to optimize.
Sartorius offers PD solutions for high productivity, flexible upstream intensification processes and increased speed to IND.
High throughput screening tools allow for faster selection of optimized clone, media and upstream process.
Be “right first time” by integrating PAT and data analytic tools for enhanced molecule characterization and process understanding.
Bioprocessing 4.0 starts from PD. Data management, trend monitoring and recipe transfer tools ensure reliable scale-up and tech transfer.
PD Process Intensification Workflow
Cell Line & Media Development
For the development of a new therapeutic, early identification of robust, stable and highly productive cell line, media and cell culture conditions suitable for intensified processes are very important. However, the multitude of time- and labor-intensive experiments required to generate a viable cell line result in longer development times and higher cost.
Early identification of a productive cell line depends on a robust and efficient screening platform or a reliable partner that can develop a perfusion process in less time. In particular, you need:
- Automated high-throughput tools mimicking perfusion process
- Simple and easy-to-use DOE software
- Expertise in cell line development and perfusion clone selection
Sartorius supplies state-of-the-art, industry-leading solutions and services for cell line development, as well as clone and media screening. Our solutions include:
- Ambr® 15 HT multi-parallel bioreactor system for efficient clone and perfusion media screening under perfusion-mimicking conditions at the smallest scale
- 4Cell® XtraCHO Media System for effective cultivation of CHO cells
- MODDE® software for straightforward DOE analysis of process parameters and their interaction effects
- Expert Cell Line Development Services using Cellca CHO Expression Platform for rapid generation of cell cultures, going from DNA to RCB in just 14 weeks
USP PD & Clarification
Accelerating upstream process development and time to IND requires a reliable, scalable and optimized process. However, manual and time-intensive development processes, lack of in-line process sensors and high-capacity clarification solutions extend development timelines.
To realize the benefits of intensified upstream process development, you need reliable and efficient scale-down models that predict performance at large scales, with the following capabilities:
- Automated high-throughput tools for perfusion
- Scale-down, perfusion-enabled benchtop bioreactors
- In-line PAT tools for better process understanding
Sartorius provides scalable and perfusion-enabled solutions for upstream process optimization and clarification from 250 mL to 2000 L, including:
- Ambr® 250 Perfusion multi-parallel bioreactor system with integrated perfusion pumps and automated bleed control for rapid perfusion process development
- Biostat® B-DCU scale-down benchtop system integrated with BioPAT® tools and MFCS software for perfusion recipes enables robust process characterization
- Sartoclear® high-capacity depth filters for robust clarification
- Sartopore® 2 sterile filters for efficient bioburden reduction while maintaining sterility
Capture Chromatography
Affinity chromatography captures the target molecule by binding it to a ligand while most of the bulk impurities flow through from the clarified feed stream. However, higher product titers obtained using perfusion processes and the labor-and time-intensive chromatography process development lead to high costs, low productivity and low product yields.
Your affinity capture program requires semi-continuous capture capabilities that can be easily integrated with upstream perfusion technology and that supports:
- Multi-column chromatography
- Flexibility and automation
Sartorius offers the following flexible and cost-effective solution for affinity chromatography, providing 3-5x productivity along with significant savings on resin costs:
- BioSMB PD System single-use, automated, high-efficiency multi-column chromatography solution for easy integration with both fed-batch and perfusion processes
Polishing
Polishing chromatography occurs post-capture to remove remaining impurities, such as HCP, DNA and potential viruses, from the product stream. However, the low capacity of chromatography media combined with highly manual and long development processes increase overall costs and extend timelines.
For effective polishing, you need scalable chromatography media that enable efficient impurity removal and that feature:
- High-efficiency and cost-effective media
- Flexibility, automation and low footprint
Sartorius provides flexible choices between membrane adsorbers and resins that reduce footprint and buffer usage, driving cost-effectiveness while increasing flexibility in operations:
- Sartobind® single batch use, high-capacity and high-flux membrane adsorbers for rapid purification of biomolecules
- CMM Hypercel™ high-efficiency and cost-effective mixed-mode resins for cation exchange chromatography
Virus Clearance
Viral clearance strategies include virus inactivation and removal. pH adjustment after low pH virus inactivation can lead to a lot of aggregate formation that requires efficient filtration. Robust removal of all potential viruses is also critical. However, low depth filter flux and capacity, low virus filter throughput and early virus breakthrough lead to long filtration times and large filter areas.
Your virus clearance strategy requires robust and scalable viral clearance and bioburden reduction solutions that deliver:
- High-capacity virus filtration
- Efficient bioburden reduction
- High product yield
- Solutions for both in-process holds and buffer solutions
Sartorius provides efficient, reliable and cost-effective solutions for viral clearance, aggregate removal and bioburden reduction:
- Sartoclear® single-use depth filters for high-capacity clarification
- Sartopore® 2 single-use sterile filters for high-capacity bioburden reduction
- Virosart® Max & Virosart® HF for robust virus prefiltration and filtration
- Flexsafe® Bags, including single-use bags and connectors, for fluid storage, mixing and in-process use
TFF & Final Filtration
Final steps of downstream processing call for tangential flow filtration (TFF) to obtain the desired product concentration and buffer formulation, as well as sterile filtration to ensure sterility and safety. However, multiple experiments for optimized formulation buffer, variation in feed streams and low flux filters result in long filtration times and large filter areas.
To produce a final drug substance, you need robust and efficient solutions for formulation screening, crossflow process development and sterile filtration of the drug substance, including:
- Automatic, high-throughput tools with DOE capability
- Low-adsorption and high-flux filters
- Optimized solutions for different feed streams
Sartorius provides robust, scalable, efficient TFF and final filtration solutions that accelerate process development and enable high product yield:
- Ambr® Crossflow high-throughput crossflow system for efficient formulation screening studies
- Sartoflow® Smart modular, flexible benchtop crossflow system for process development
- Hydrosart® Cassettes with extremely hydrophilic, low-adsorptive and high-flux membranes for TFF
- Sartopore® Platinum single-use sterile filters for final filtration of drug substance
Cell Line Development and Clone Selection
PI Process Compatibility Starts Here
Cell Line Development (CLD) includes gene cloning, cell line selection and characterization, media selection and Master Cell and Working Cell banking. Many regard the selection of the appropriate production clone as the most critical step in CLD. If the PD team starts with one clone and then decides to switch to another, it would require major process changes . These changes would bring both significant delays and additional costs.
This is why development and characterization of the cell line for identity, purity and stability are crucial. Sartorius is a reliable partner for these critical steps, offering a Cell Line Development Service, a high-titer cell line with CellcaCHO Expression Platform, and perfusion enabled media.
Cell Line Development Service
The Intensification Potential with CellcaCHO
The CellcaCHO Expression Platform is a scalable cell line and fed-batch process development system. It delivers stable research clones from DNA to research cell bank in 14 weeks and a fully characterized master cell bank in 10 months.
The CellcaCHO supports the potential of optimization:
- Combine with N-1 perfusion to more than double titer with 30-40% fewer culture days in the production bioreactor, resulting in up to 2x productivity
- Use in concentrated fed-batch to enable a double-digit titer resulting in up to 4x productivity
- Pair with dynamic perfusion to achieve a 5x increase in productivity
Cell Culture Media
Chemically Defined, Optimized and Highly Productive
The 4Cell® XtraCHO-Media system is chemically defined and optimized for CHO DG44 cell lines:
- Contains CHO media and feeds, optimized for protein manufacturing
- Supports rapid growth to high densities and high cell growth rates compared to standard-serum supplemented media
- All the critical parameters (e.g., viability, titer and lactate) remain consistent while cultivating the cells in bioreactors with various volumes
- Processes can be easily transferred and scaled-up to bioreactors up to 2,000 L, saving up to 3 months for scalability studies
The 4Cell® XtraCHO-Media system is cost-efficient, compliant with regulatory guidelines, and provides reliable batch-to-batch consistency.
Intensify to Meet Current and Future Demands
Clone and Media Optimization
Optimization for High Productivity
Cell culture media optimization is a crucial point of focus when exploring perfusion processes to achieve higher productivity. The medium should support rapid growth to high densities, achieve high cell growth rates with maximum viable cell concentration and provide batch-to-batch consistency.
PD Optimization Services
Process Development Services for Perfusion Process Optimization
Many cell lines were identified in a standard fed-batch process performed in Ambr® 15 microbioreactors. These cell lines show high intensification potential, but the best clone from standard fed-batch selection might not be the best in perfusion.
Mimicking the perfusion process for clone selection will not only save time, it can also enable a three-fold increase in productivity. Sartorius‘ Process Development Service makes this possible by offering a combined solution that includes cell line, media, process development and the technology to deliver optimal results.
Ambr® 15 Bioreactor
Rely on Ambr® 15 to Deliver the Data Needed for Intensification
The Ambr® 15 is the smallest single-use bioreactor on the market with system automation and parallel operation, and can be a starting point for process intensification:
- Identify suitable clones, media and feeds to achieve great performance in perfusion mode
- Mimic perfusion process to produce reliable and relatable data for intensified processes
- Reduce cost-per-experiment by saving substantial amounts on facility space, capital, labor, media and consumables
MODDE® DOE Software
An Easier Path to DOE
MODDE® is an easy-to-use solution for Design of Experiments (DOE). It offers design space functionality, multiple automated workflows, and improved process understanding through visualization and graph interaction. With an efficient DOE approach to problem-solving, you can:
- Significantly reduce experimental costs
- De-risk projects and increase success rates
- Make the most of valuable samples, raw materials and human resources
- Accelerate progress and time-to-market while keeping within budget
- Achieve quality goals and satisfy Quality by Design requirements
Advanced Flow Cytometry
iQue3 for Actionable Results from Smaller Samples in Record Time
iQue3 is an advanced flow cytometry platform for high throughput clone screening.
- Rapid microvolume sampling provides the fastest sample acquisition in the industry
- 96-well plates processed in as few as five minutes
- 384-well plates in 15-20 minutes
- Requires the smallest assay volumes and sample volumes – as little as a 1 mL – saving reagents and conserving precious cells
- Key applications for iQue3 with process intensification are clone ranking and selection of production clones with rapid and simultaneous evaluation of IgG titer, specific productivity, and cell health
Intensify to Meet Current and Future Demands
Process Characterization
Process Characterization for Establishing Perfusion-Based Processes
Characterizing an established perfusion-based process in scale-down models helps identify the economy of the process while mimicking the performance of large-scale bioreactors. Process parameters such as cell density and viability, titer and lactate level must be monitored in real-time to get the most benefit from the data.
Sartorius offers single-use and scalable technology solutions to supports perfusion-based process development.
Biostat® B-DCU
Biostat® B-DCU Benchtop Bioreactor
The Biostat® B-DCU benchtop bioreactor with advanced PAT and MFCS perfusion recipes can be used from screening to characterization of intensified processes.
- Configurable options offer maximum flexibility
- Accommodates working volumes from 250 mL to 10 L
- Aeration and feeding strategies with sensor technologies mimic commercial-scale bioreactors
- Configurable mass flow controllers cover a flow range from 1 mL to 20 L per minute
- Expand existing system by adding further supply towers (up to 6)
Ambr® 250 HT
A Fully Integrated Solution for Perfusion at PD Scale
Ambr® 250 HT perfusion supports the rapid development of scalable perfusion processes:
- Up to 24 x 100-250 mL single-use perfusion bioreactors operating in parallel
- Includes a fully automated liquid handling platform
- Reduces systems hands-on time by 50%
- Requires 13% less media than benchtop bioreactors for lower experiment costs
Only Sartorius offers a fully integrated solution for perfusion at the PD scale. Processes developed in Ambr® systems can be easily scaled from RM 20/50 to RM 200 and up to STR 2000 L.
Octet Systems
Octet Systems for Functional Characterization and Protein Quantitation
Octet is a low maintenance, label-free biomolecular interactions analysis platform. It monitors binding of proteins from solution to a biosensor surface in real time. Using Octet during process characterization effectively:
- Reduces experiment time: 96 samples in 20 minutes, 384 samples in 70 minutes
- Delivers 20x faster than traditional ELISA and HPLC (IgG titer)
- Operates 2x faster than traditional SPR (method development)
- Increases capacity for QC samples testing
- Only requires 10% the hands-on time of traditional ELISA
Key applications for process intensification are functional characterization and protein quantification, as well as glycan screening and contaminant testing. Octet can also be used for residual host cell protein detection.
