Temperature Shift for Selective Antibody De-cysteinylation
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Solution Overview
Problem
Current methods for selectively reducing unpaired cysteines in monoclonal antibodies, such as secukinumab, require strict oxygen level control and additional steps, leading to increased costs and complexity in large-scale production, while maintaining the integrity of conserved disulfide bonds.
Innovation Solution
A method involving temperature shift during cell culture to selectively reduce unpaired cysteines without the need for reducing agents, either by culturing cells at a lower temperature or using on-column reduction with chromatography, optimizing pH and duration to achieve high de-cysteinylation levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If reducing agents such as cysteine are added to the antibody mixture to reduce unpaired cysteines, then de-cysteinylation of unpaired cysteines is improved, but process complexity and cost increase due to additional steps and strict oxygen level control requirements
Solution Approach 1:
The patent extracts and removes the need for reducing agents from the process. By using temperature shift during cell culture, the method achieves de-cysteinylation of unpaired cysteines without adding cysteine or other reducing agents, thereby eliminating the complexity associated with their addition, monitoring, and removal.
Solution Approach 2:
The patent enables the system to perform de-cysteinylation autonomously through temperature shift during cell culture. The cells themselves, when cultured at lower temperatures, naturally produce antibodies with reduced unpaired cysteines, eliminating the need for external reducing agents and additional processing steps.
2Manufacturing precision
If reducing agents are added to reduce unpaired cysteines, then de-cysteinylation is improved, but production cost increases due to additional reagents and process steps
Solution Approach 1:
The patent removes the requirement for reducing agents such as cysteine from the manufacturing process. By achieving de-cysteinylation through temperature shift during cell culture alone, it eliminates the cost of purchasing, storing, and handling reducing agents as well as the costs associated with additional process steps.
Solution Approach 2:
The system uses the cells' natural response to temperature shift to achieve de-cysteinylation. This self-service mechanism eliminates the need for expensive external reagents and complex additional processing steps, thereby reducing overall production costs.
3Manufacturing precision
If strict oxygen level control is maintained during reducing agent treatment, then selective reduction of unpaired cysteines is improved, but process complexity and operational difficulty increase
Solution Approach 1:
The patent extracts and eliminates the requirement for strict oxygen level control from the process. By using temperature shift during cell culture to achieve de-cysteinylation, it removes the need for complex oxygen monitoring and control systems that are required when using reducing agents.
Solution Approach 2:
The method relies on the cells' natural metabolic response to temperature shift to achieve selective de-cysteinylation. This self-regulating mechanism eliminates the need for external oxygen level control, simplifying operational procedures and reducing the skill level required for process execution.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach results in stable, biologically active monoclonal antibodies with high de-cysteinylation levels, reducing production costs and simplifying the process by eliminating the need for additional steps and oxygen level control.
Implementation Method 1
A method involving temperature shift during cell culture to selectively reduce unpaired cysteines without the need for reducing agents, either by culturing cells at a lower temperature
Data Source
AI summary
The invention relates to upstream and/or downstream processes of selectively reducing one or more unpaired cysteines of a monoclonal antibody, whilst keeping conserved inter- and intra-molecular disulfide bonds elsewhere in the antibody intact. The invention further relates to purified antibodies obtained by the methods as described herein.