Two-Step Bis-Tris Chromatography for Monoclonal Antibody Purification
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Solution Overview
Problem
The conventional three-step chromatography process for monoclonal antibody purification is time-consuming and labor-intensive, requiring multiple buffer exchanges and manual interventions, which increases the risk of contamination and limits scalability and efficiency in producing large batches quickly.
Innovation Solution
A two-step chromatography process using affinity chromatography and multi-modal resin chromatography with buffers composed of Bis Tris, NaCl, and acetic acid, allowing for automated continuous operation without buffer exchange, reducing the number of steps and manual interventions, and enabling rapid production of high-purity antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If a conventional three-step chromatography process is used for monoclonal antibody purification, then the purification can be completed with adequate purity, but the process time is extended to 3-5 working days and requires multiple manual interventions
Solution Approach 1:
The patent combines the capture step and ionic exchange step into a single multi-modal resin chromatography step that performs both functions simultaneously. This merging of steps reduces the total number of chromatography steps from three to two, thereby reducing process time while maintaining purification purity through the integrated resin design that provides both affinity capture and ionic exchange capabilities in one column
Solution Approach 2:
The patent implements continuous mode operation where the crude protein eluent from the first chromatography step is directly passed to the second chromatography step without interruption, storage, or manual buffer exchange. This continuous flow eliminates idle time between steps and removes the need for open phases, reducing the overall process time from 3-5 days to a more rapid continuous process while maintaining adequate purification through uninterrupted processing
2Manufacturing precision
If multiple buffer exchanges and manual interventions are performed during chromatography steps, then the purification process can be completed, but the risk of contamination increases and automation becomes more difficult
Solution Approach 1:
The patent eliminates buffer exchanges between chromatography steps by implementing continuous mode operation where the crude protein eluent flows directly from the first to the second chromatography column. This continuous process removes the need to open the system for buffer changes, thereby eliminating contamination risks associated with manual interventions while maintaining purification purity through the integrated multi-modal resin design that handles both capture and ionic exchange in a closed system
Solution Approach 2:
The patent uses the crude protein eluent itself as an intermediary that directly connects the first and second chromatography steps without requiring buffer exchange. The eluent serves as the continuous medium that carries the purified protein from the capture step through the ionic exchange step, eliminating the need for intermediate buffer solutions and manual interventions that would increase contamination risk
3Manufacturing precision
If three separate chromatography steps are used for capture, ionic exchange, and polishing, then adequate purification can be achieved, but the device complexity and number of manual operations increase
Solution Approach 1:
The patent merges the capture step and ionic exchange step into a single multi-modal resin chromatography column that performs both functions simultaneously. This reduces the total number of chromatography steps from three to two, simplifying the device complexity and reducing the number of manual operations required while maintaining purification purity through the integrated resin design that provides both affinity capture and ionic exchange capabilities in one column
Solution Approach 2:
The patent employs a multi-modal resin that performs multiple functions within a single chromatography column, including affinity capture and ionic exchange. This multi-functional resin eliminates the need for separate dedicated columns for each function, reducing device complexity and the number of steps required while achieving adequate purification through the combined capabilities of the universal resin
4Manufacturing precision
If conventional buffer systems are used requiring different chemicals for each step, then the purification can be completed, but the time for preparing chromatographic columns increases and manual interventions are required
Solution Approach 1:
The patent uses a universal buffer system based on Bis Tris that can be used for all chromatography steps including equilibration, washing, and elution. This single buffer system replaces the conventional requirement for multiple different buffer solutions, simplifying buffer preparation and reducing the time required to prepare chromatographic columns while maintaining purification purity through the appropriate pH adjustments and salt concentrations within the universal buffer
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 significantly reduces the time and cost of antibody purification, enhances yield and purity, and facilitates scalability by eliminating the need for manual buffer changes and open phases, making it suitable for industrial-scale production.
Implementation Method 1
one affinity chromatography
Implementation Method 2
one multi-modal resin chromatography
Data Source
AI summary
The invention provides a two-step chromatography process for small and large-scale purification of proteins, specifically monoclonal antibodies, using only four buffer solutions made from a mother solution.