Overload Elute Chromatography for High-Load Polypeptide Purification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
The large-scale, cost-effective purification of polypeptides, such as monoclonal antibodies, is challenging due to low dynamic binding capacity of existing chromatography resins, limiting plant throughput and requiring large columns or multiple cycles, especially for non-platform MAbs.
Innovation Solution
A method involving overload and elute chromatography (OEC) is employed, where a composition exceeding the dynamic binding capacity of the chromatography material is loaded, followed by elution under specific conditions to separate the polypeptide from contaminants, utilizing resins like mixed mode, anion exchange, hydrophobic interaction, or affinity materials.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If bind and elute chromatography is used to maximize dynamic binding capacity, then product purity is improved, but load density is restricted and plant throughput is limited
Solution Approach 1:
The patent inverts the traditional bind-and-elute approach by using flow-through mode where impurities bind to the chromatography resin while the product flows through. This reversal allows high load density operation without compromising product purity, as the selectivity is achieved by binding contaminants rather than the product of interest.
Solution Approach 2:
The patent changes the binding parameters by adjusting pH and ionic strength conditions to create selective binding of impurities to the resin. By modifying these parameters, the system achieves enhanced purification capacity and allows operation at load densities exceeding traditional DBC limitations while maintaining product purity.
2Productivity
If flow through chromatography is used for high load density, then productivity is improved, but implementation is limited for non-platform MAbs and existing manufacturing plants
Solution Approach 1:
The patent creates a universal chromatography approach that can handle both platform and non-platform MAbs using the same flow-through mode. The method is adaptable to existing manufacturing plants because it uses standard chromatography resins and equipment, eliminating the need for specialized implementations while achieving high load density for diverse antibody types.
3Manufacturing precision
If weak partitioning chromatography is used to enhance purification, then product purity is improved, but load conditions are restricted to low partition coefficient range
Solution Approach 1:
The patent introduces dynamic load conditions that are not restricted to a fixed low partition coefficient range. By using flow-through mode with adjustable pH and ionic strength, the system can dynamically optimize binding conditions for different antibody types and impurity profiles, enhancing purification while maintaining load condition flexibility.
4Productivity
If overload chromatography is used beyond dynamic binding capacity, then productivity is improved, but product yield decreases due to lack of elution phase
Solution Approach 1:
The patent extracts the product of interest from the bound impurities by using flow-through mode where the product does not bind to the resin and passes through in the effluent. This extraction approach achieves high productivity by loading beyond DBC while maintaining high yield, as the product is recovered in the flow-through fractions without requiring an elution phase that would otherwise be needed to release bound product.
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 enhances chromatography material utilization, allowing for smaller columns, improved plant fit, and lower costs by achieving significant impurity removal and increased yield, with polypeptide purity and efficiency improved through pooling and elution strategies.
Implementation Method 1
loading the composition onto a chromatography material in an amount in excess of the dynamic binding capacity of the chromatography material for the polypeptide
Implementation Method 2
eluting the polypeptide from the chromatography material under conditions wherein the one or more contaminants remain bound to the chromatography material using an elution buffer, wherein the elution buffer has a conductivity less than the conductivity of the loading buffer
Data Source
Figure 1a~1c
Figure 2
Figure 3
AI summary
The present invention provides methods for purifying a polypeptide from a composition comprising the polypeptide and at least one contaminant by overloading a chromatography material and eluting the product.