The antibody manufacturing process, regardless of the specific antibody like ustekinumab, can be broadly divided into upstream and downstream processing:
Upstream Processing:
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Cell Line Development:
- Isolate specific B cells producing the desired antibody from immunized animals or humanized systems.
- Create immortalized cell lines (hybridoma or recombinant) that stably produce the antibody.
- Optimize cell line growth and antibody production in bioreactors.
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Cell Culture and Fermentation:
- Large-scale cultivation of the selected cell line in controlled bioreactors with optimal conditions for efficient antibody production.
- Monitoring and adjusting various parameters like nutrients, pH, temperature, and oxygen levels.
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Harvest and Clarification:
- Harvesting the cell culture broth containing the produced antibody.
- Removing cellular debris and other impurities through centrifugation and filtration.
Downstream Processing:
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Capture and Purification:
- Employing chromatography techniques to capture the target antibody with high specificity and purity.
- Sequential steps like protein A chromatography, ion exchange, and size exclusion chromatography.
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Concentration and Formulation:
- Concentrating the purified antibody solution to the desired level.
- Adding stabilizing excipients (salts, sugars) and buffers to maintain stability and functionality.
- Sterile filtration and filling into vials or syringes.
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Quality Control:
- Extensive testing to ensure the final product meets stringent quality standards.
- Purity, potency, activity, and absence of contaminants are rigorously assessed.
Critical Steps and Parameters:
- Cell line stability and performance: Maintaining consistent antibody production throughout the manufacturing process.
- Bioreactor conditions: Precise control of pH, temperature, nutrients, and oxygen levels for optimal cell growth and antibody production.
- Purification efficiency: Minimizing losses while ensuring high purity and specificity of the target antibody.
- Sterility and control of contaminants: Preventing contamination throughout the process to ensure a safe and pure product.
- Process consistency and scalability: Establishing robust and reproducible protocols for large-scale manufacturing.
It's important to note that this is a simplified overview, and the specific steps and parameters involved may vary depending on the specific antibody and desired characteristics. Additionally, advancements in technologies are constantly evolving, influencing the manufacturing process for improved efficiency and quality.
Picture Credit: AstraZeneca
