Case study - downstream processing of monoclonal antibodies produced in bioreactors

Summary

This video delves into the process of downstream processing for monoclonal antibodies, focusing on their increasing importance in treating diseases. The speaker highlights key aspects, such as the need for stability during harvesting, the role of protein A Affinity chromatography, and the significance of inline process monitoring using sensors. The discussion touches on the shift towards continuous processing and single-use technologies, cost-effective alternatives to protein A, and quality control methods throughout the process. The speaker also emphasizes the growing relevance of non-animal technologies and provides a comprehensive look at the steps involved in monoclonal antibody production.

Takeaways

• 😀 Monoclonal antibodies (mAbs) are crucial in treating diseases, especially autoimmune disorders, and are becoming more prevalent in the pharmaceutical industry.
• 😀 There is a growing trend toward continuous bioreactors over batch processes because they are more cost-effective for large-scale production.
• 😀 Single-use technologies like biowave reactors are increasingly adopted for flexibility, though they raise concerns about plastic waste.
• 😀 Process Analytical Technologies (PAT) and Quality by Design (QbD) are vital in ensuring product quality by monitoring critical quality attributes (CQAs) throughout production.
• 😀 Protein A Affinity Chromatography is commonly used to purify mAbs but is costly due to expensive resin, accounting for more than 50% of downstream processing costs.
• 😀 Alternatives to Protein A chromatography, such as multimodal chromatography, are being explored to reduce costs without compromising purity.
• 😀 Key stability factors in mAb production include pH, temperature, shear stress, and colloidal stability, which must be carefully controlled during harvesting and processing.
• 😀 Computational modeling and in silico tools are used to predict stability and optimize production processes, speeding up development and improving efficiency.
• 😀 Viral clearance steps, including filtration and chromatography, are essential for ensuring the safety and sterility of mAbs during processing.
• 😀 Ethical considerations drive the use of non-animal technologies for antibody production, with a focus on refinement, reduction, and replacement of animal models in research.

Q & A

What are monoclonal antibodies, and why are they important in the pharmaceutical industry?

Monoclonal antibodies are highly specific antibodies produced to target particular antigens, which are used to treat diseases. They are important in the pharmaceutical industry because over 500 monoclonal antibodies are already in use for various treatments, including autoimmune diseases, and they play a crucial role in medical therapeutics.

How do monoclonal antibodies relate to the body's immune response?

Monoclonal antibodies mimic the body's natural immune response by recognizing and binding to foreign invaders, such as viruses or bacteria, thereby neutralizing them. In the body, antibodies are naturally produced when a foreign substance enters, and monoclonal antibodies function similarly to help fight disease.

Why is the stability of monoclonal antibodies important during the harvesting process?

Stability is critical because antibodies can be sensitive to various environmental factors like pH, temperature, and shear stress during the harvesting process. Ensuring stability ensures that the antibodies retain their effectiveness and are not damaged during production and purification.

What role do computational models and in-silico tools play in antibody production?

Computational models and in-silico tools help predict the behavior of monoclonal antibodies during production, which speeds up the process. These tools allow for a more efficient approach by simulating potential modifications and stability issues, helping improve the overall yield and quality.

What are the key trends in the bioreactor and downstream processing for monoclonal antibodies?

Key trends include a shift from batch to continuous reactors for cost-effectiveness, the use of smaller single-use reactors in response to market demands (especially post-pandemic), and the increased use of Process Analytical Technologies (PAT) for real-time monitoring during the process.

How does Process Analytical Technology (PAT) benefit monoclonal antibody production?

PAT benefits monoclonal antibody production by enabling real-time monitoring of critical process parameters, such as pH, conductivity, and protein concentration. This helps ensure consistent product quality and reduces the need for post-production testing.

What is Protein A Affinity Chromatography, and why is it commonly used in antibody purification?

Protein A Affinity Chromatography is a technique where Protein A binds specifically to the Fc region of antibodies, allowing them to be purified with high selectivity and speed. It is commonly used because it effectively removes impurities, such as host cell proteins and DNA, resulting in high-purity antibodies.

What are the disadvantages of using Protein A Affinity Chromatography in monoclonal antibody production?

The main disadvantage of Protein A Affinity Chromatography is the high cost of the resin, which can account for up to 50% of the downstream processing costs. Additionally, some antibodies may not tolerate the low pH conditions required for elution, leading to potential instability or product loss.

What alternative techniques are being researched to replace Protein A Affinity Chromatography?

Alternatives being researched include membrane flotation, crystallization steps, and multimodal chromatography. These alternatives aim to lower costs and reduce the harsh conditions that might affect antibody stability.

How does glycosylation affect monoclonal antibody production and heterogeneity?

Glycosylation, the process of adding sugars to the antibody, can vary during production, leading to heterogeneity in the final product. Differences in glycosylation patterns can impact the stability, efficacy, and safety of monoclonal antibodies, which is why it’s critical to control production conditions and ensure consistency.