Optimize your process from cell line to high-throughput screening
Monoclonal antibodies (mAbs) and their variants, such as bispecific antibodies and antibody drug conjugates (ADCs), continue to be promising choices for creating new treatments for cancer, autoimmune diseases, and other chronic conditions. Companies must focus on strong process development practices as they strive to produce these therapies quickly and efficiently. This includes creating stable CHO cell lines, selecting the best media and feed, and speeding up downstream development with high-throughput screening techniques. Therefore, it is important to consider the tools available to improve every stage of mAb process development.
Facilitate stable cell line development for mAb production
Strategic cell line development (CLD) can facilitate achieving high titers, rapid timelines, regulatory compliance, and low licensing costs. Gibco Freedom CLD kits each come with a cGMP-banked cell line provided in an animal origin free, chemically defined medium, enabling fast transition from development to clinical trials and commercial production. Thermo Fisher Scientific offers flexible commercial-use licensing for all Freedom CLD kits.
The Freedom CLD kits have been proven to help achieve immunoglobulin G (IgG) titers of 3–6 g/L while enabling clients to move from transfection to identifying top-producing clonal cell lines in as little as four months. When comparing our two Freedom CLD kits—DG44 and CHO-S—the primary difference is that DG44 cells undergo genetic amplification of the gene of interest following treatment with methotrexate (MTX) due to their deficiency in the dihydrofolate reductase (DHFR) gene. DHFR is provided exogenously with the pOpticVEC vector. Because of this amplification step, the timeline for generating stable clones with the DG44 kit is 36 weeks, while the timeline for generating stable clones with the CHO-S kits is approximately 24 weeks.
Leverage optimized media feed and diversity
The Gibco Efficient-Pro System offers a specific catalog of media and feed combinations designed with versatility, performance, and scalability in mind with proven capability to produce mAbs, bispecific antibodies, and novel protein therapeutics. To facilitate process optimization and scalability, these formulations are nutrient-rich to support higher cell density and titer.
In a study to evaluate the benefits of Efficient-Pro Feed with two different cell lines, Efficient-Pro Feed 2 was tested at 1.5% to 3% compared to a commercially available medium and feed process. Overall, when Efficient-Pro Feed 2 was supplemented at 1.5% daily, it achieved 3x the titer compared to the other supplier’s medium and feed. In another study that assessed viable cell density (VCD), percent viability, and titer profiles, Efficient Pro Media and Feed were shown to support consistent CHO-K1 cell growth and productivity across different bioreactors.
CHO media and feed panel libraries can be leveraged to screen diverse formulations with different cell lines to improve cell growth, titer, and/or product quality. Customers can rapidly review Thermo Fisher’s libraries to identify the best options, and if an improved alternative is discovered, formulations are available, ready for shipment, and scalable to cGMP standards. Additionally, Thermo Fisher can provide customers with a sample study protocol, field application scientist (FAS) consultation, and support from knowledgeable technical teams.
When evaluating a panel, it is important to consider a range of formulations to accommodate the unique metabolic requirements of your clone(s) and optimize efficiency and cost-effectiveness. Following a media panel evaluation, a customer might choose to test the feed panel to determine the optimal media feed combination. Once chosen, this formulation can be scaled up via Thermo Fisher’s Gibco Rapid Prototyping (GRP) or cGMP manufacturing facility. If improvements are needed, spent media analysis can be performed to determine an appropriate feed strategy to test or modify through the GRP facility. If a customer wants to identify a lead formulation and confirm results in a second round of screening, Thermo Fisher can provide up to 5 L of the lead medium or 500 mL of the lead feed.
Select the appropriate resin for your downstream process
Affinity and polishing resins are two critical components of mAb downstream process development. Resins can be a major tool in alleviating the inherent challenges of purification for increasingly diverse mAb therapeutics. CaptureSelect and POROS resin technologies aim to address today’s challenges in the purification workflow and include the following:
- CaptureSelect and POROS affinity resins: High-performance affinity resins to support mAb therapeutics, viral vectors, mRNA, proteins, and vaccines. mAb-related options include MabCaptureC, CaptureSelect CH1-XL, CaptureSelect KappaXP, CaptureSelect LambdaXP, and CaptureSelect FcXP.
- POROS ion exchange resins: Performance independent of flow rate, high capacity, and salt tolerance. POROS offers four different ion exchange resins: two weak exchangers (D50 and PI50) and two strong exchangers (HQ50 and XQ).
- POROS hydrophobic interaction resins: Selectivity through a wide range of hydrophobic conditions. There are three hydrophobic resins available:
- POROS Ethyl, which works in bind/elute mode of moderately hydrophobic molecules.
- POROS Benzyl, which works in bind/elute mode or flow-through mode depending on the hydrophobicity of the molecule
- POROS Benzyl Ultra, which works in flow-through mode in lower salt concentration
- POROS Caprylate Mixed Mode Cation Exchange Chromatography Resin: High aggregate selectivity, increased productivity, and high scalability through flowthrough application
Conduct high-throughput screening
High-throughput screening methods allow developers to generate massive amounts of data through miniaturization and parallelization of experiments. When time and material are limited, high-throughput screening allows multiple experiments to be run simultaneously to reduce time waste. Typical chromatography process development starts with high-throughput screening with miniaturized systems and the ability to experiment with multiple variables at the same time.
Resin slurry plates and miniature columns are two common approaches to high-throughput screening. Resin slurry plates allow for more conditions to be run in parallel with lower capital investment and a broader window of operation. They can be easier to operate and require less material in terms of resin and feedstock. These plates can either be self-made or purchased prefilled while miniature columns require the use of pre-paid products.
Choose what works for you
In the pursuit of making your mAb process as efficient as possible, it is vital to identify the cell line, media, feed, and resin that best suit your productivity goals. Selecting the optimal components for your workflow can help yield major benefits in terms of cost savings, efficiency, and therapeutic quality.
Find more solutions on mAb process development >
Contact us for more information >
Leave a Reply