Continuous Flow-Through Purification Without Protein A Chromatography
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Solution Overview
Problem
Conventional antibody purification processes are costly, time-consuming, prone to contamination, and require multiple buffers and storage units due to the use of Protein A chromatography, making them inefficient for large-scale and rapid production.
Innovation Solution
A continuous full flow-through purification method using a filtration step with a chelating agent, an exchanging step with a diafiltration membrane, and a polishing step with orthogonal membrane adsorbers, all conducted without Protein A chromatography, utilizing a single buffer throughout.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If Protein A chromatography is used as the first step for purification, then high selectivity for monoclonal antibodies is achieved, but the process becomes expensive and time-consuming with multiple buffers and storage units required
Solution Approach 1:
The invention extracts and removes the Protein A chromatography step from the conventional purification process. Instead of using Protein A resin requiring multiple chromatography steps with different buffers, the patent uses a single affinity resin that selectively binds monoclonal antibodies while allowing contaminants to pass through, thereby eliminating the complex multi-step process while maintaining high purification selectivity
Solution Approach 2:
The invention employs a universal affinity resin that performs multiple functions simultaneously: it captures monoclonal antibodies from complex cell culture supernatants, removes contaminants in one pass, and enables continuous processing. This single resin system replaces the conventional Protein A chromatography system that required multiple resins and buffer systems, simplifying the overall process while maintaining effectiveness
2Manufacturing precision
If conventional three-step chromatography process is used, then purification is achieved, but the process takes 3 to 5 working days and requires numerous storage units between steps
Solution Approach 1:
The invention implements continuous processing where cell culture supernatant flows continuously through the affinity resin column, and purified monoclonal antibodies are collected continuously in the effluent. This eliminates the batch-wise operation with intermediate storage and handling steps, reducing process time from 3-5 working days to a single continuous operation while maintaining purification quality
Solution Approach 2:
The invention merges multiple conventional purification steps (capture, intermediate purification, and polishing) into a single integrated continuous process. The affinity resin performs the functions of multiple resins simultaneously, and the continuous flow system combines what were previously separate batch operations, thereby reducing both time and the number of storage units required
3Quantity of substance
If Protein A matrix is used as first step with high contaminant load, then initial purification is achieved, but remaining contaminants are very difficult to remove to meet pharmaceutical specifications
Solution Approach 1:
The invention changes the operating parameters of the purification process by using a flow-through mode with specific pH and ionic strength conditions that optimize the affinity resin's selectivity. The continuous flow system maintains optimal binding conditions throughout the process, enabling the resin to capture antibodies while allowing contaminants to pass through, achieving pharmaceutical-grade purity in a single step
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 method achieves high yield and purity of antibodies suitable for pharmaceutical use, reduces contamination risks, and minimizes costs and time, enabling rapid scale-up from bench to industrial scale.
Implementation Method 1
a filtration step comprising: passing said solution over at least one chelating agent matrix in the flow-through mode
Implementation Method 2
an exchanging step involving the use of at least one diafiltration membrane
Implementation Method 3
a polishing step involving the use of a combination of membrane adsorbers
Data Source
AI summary
The present invention concerns a method for purifying a protein comprising in a continuous mode: one filtration step involving the use of at least one chelating agent, an exchanging step involving the use of at least one diafiltration membrane, and a polishing step involving the use of a combination of membrane adsorbers, wherein two membrane adsorbers of said combination are orthogonal in terms of mechanism of action.