Introduction
Bispecific monoclonal antibodies (bispecific mAbs) are an innovative class of therapeutic proteins designed to simultaneously target two different antigens or epitopes, which may result in multiple or synergistic biological effects. The development of bispecific mAbs involves intricate processes to ensure high process recovery, product purity, and overcoming challenges unique to this field. In this blog post, we'll delve into key considerations in developing bispecific mAbs, highlighting our purification process optimization.
Typical Challenges in Bispecific mAb Development
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Choosing the appropriate molecular format and engineering method for the bispecific antibody, depending on the desired properties, such as stability, solubility, expression, purification, and pharmacokinetics.
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Determining the optimal binding affinity and kinetics of each arm of the bispecific antibody for its target, as well as the potential cross-reactivity and interference with other molecules.
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Establishing the quality and manufacturing specifications for the bispecific antibody, including the product characterization, bioassay, stability, and cGMP compliance.
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Achieving high recovery without compromising the product purity
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Overcoming concentration-related aggregation, which can affect both yield and quality.
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Separating the target molecule from its similar mispairing impurities.
Case Study: Developing a downstream process for a symmetry tetravalent bispecific antibody.
In EirGenix’s pursuit of excellence in developing the most optimized process for bispecific monoclonal antibodies (mAb), we present a compelling case study featuring a symmetry tetravalent bispecific antibody. This bi-specific antibody underwent a meticulous development process with a focus on optimizing purification strategies.
Our innovative approach incorporated a unique wash step in the Protein A purification process, elevating impurity removal to unprecedented levels. To combat aggregation risks, a double flow-through mode purification process was devised, ensuring the antibody's structural integrity. Striking the delicate balance between recovery and purity, the process was fine-tuned to achieve a remarkable 60% process recovery without compromising the final purity, reaching an impressive 97.64%. The Host Cell Protein (HCP) levels were meticulously controlled, with a measured value of 4.29 ppm. Notably, even after the Protein A purification step, the antibody maintained a robust 88.88% purity.
This case study stands as a testament to EirGenix’s commitment to overcoming challenges in bispecific mAb development, showcasing our dedication to advancing the frontiers of biopharmaceutical innovation. This case study exemplifies the intersection of precision, efficiency, and excellence in therapeutic protein development.
Key Considerations in Developing a Bispecific or poly-specific mAb.
1. Achieving High Process Recovery
Achieving a high process recovery is crucial for maximizing yield. Our optimized process yields a final process recovery ranging from 42.5% to 59%.
2. Maintaining High Product Purity
Product purity is paramount to the efficacy and safety of bispecific mAbs. Our process reaches a final purity of 97.64%, with an impressive final Host Cell Protein (HCP) value of 4.29 ppm.
3. Sustaining Over 80% Purity after the First Affinity Column
Early-stage purity is essential for downstream processes. Our approach ensures over 88% purity after the first affinity column, setting the stage for subsequent purification steps.
Conclusion
Bi-specific and poly-specific monoclonal antibody development and manufacturing involves intricate processes to ensure product purity, high process recovery, and avoiding concentration-related aggregation which can lower yield. In addition, there are concerns with the mispairing of impurities that can result in separation difficulties. It is important to optimize the purification process. Further, the FDA has issued a guidance document for Sponsors that provides recommendations for the development of bispecific antibodies, covering the regulatory, quality, nonclinical, and clinical aspects. The guidance also discusses the challenges and opportunities for bispecific antibody development, such as the immunogenicity, heterogeneity, and complexity of the molecules. For deeper insights into EirGenix’s methodologies and potential collaborations, our technical team is ready to engage in further discussions with you. Join us in shaping the future of biopharmaceuticals.
Consider working with EirGenix!
Are you seeking a contract development and manufacturing partner that has it all – quality, reliability, flexibility, and reasonable pricing? At EirGenix, we offer all of this and top-notch customer service. Since 2012, EirGenix has offered mammalian and microbial biopharma development and production including cell line establishment, large-scale production process development, analytical method development, and cGMP quality system operations that are certified by the US FDA, Japan PMDA, Australian TGA, Taiwan FDA, and Europe’s EMA. You can virtually tour our site here. We would appreciate the opportunity to collaborate with you! Click on this link to contact our BD team.
Citations
The FDA bispecific guidance document for the industry
Keywords:
Bispecific Monoclonal Antibody, Bi-mAb, mAb, mammalian, bacterial, biopharmaceuticals, Appended IgG; IgM, Fab; Fc heterodimerization; bispecific antibodies; polyspecific mAb, fusion proteins; immunoglobulin; scFv; BiTE; Bispecific T cell engager.