Polymeric Mesh Purification of Macromolecules
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Solution Overview
Problem
Current chromatographic methods for large-scale purification of bio-macromolecules, such as antibodies, face limitations due to low productivity, limited capacity, and low resolution, particularly in size exclusion chromatography, which struggles with the separation of molecules based on size within a limited liquid volume, leading to inefficient recovery and purification processes.
Innovation Solution
A polymeric mesh is used for the separation of target proteins from impurities, where the mesh is designed to retain impurities with smaller molecular sizes within its pores while allowing larger target proteins to remain unbound, utilizing a cross-linked amino polymer immobilized on a porous support material, enabling efficient depletion of host cell proteins, nucleic acids, and other impurities through steric exclusion and non-covalent interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If size exclusion chromatography is used for purification, then separation based on molecular size is achieved, but productivity and loading capacity are limited due to low resolution and limited liquid volume
Solution Approach 1:
The patent employs a polymeric mesh with controlled pore size and distribution as the stationary phase. The mesh structure provides size-based separation through steric exclusion, where pores are sized to exclude target macromolecules while allowing smaller impurities to enter and be retained. This porous architecture enables high loading capacity by utilizing the entire column volume effectively, resolving the contradiction between separation resolution and productivity.
Solution Approach 2:
The patent uses composite polymeric materials combining different polymer components with specific functional groups. These composite materials provide both the size exclusion mechanism through controlled porosity and enhanced interaction sites for selective retention of impurities. The composite nature allows optimization of both separation resolution and loading capacity simultaneously.
2Reliability
If conventional chromatographic capture methods are used, then target compounds are bound selectively, but process streamlining is reduced and product recovery efficiency decreases
Solution Approach 1:
The patent inverts the conventional chromatographic approach by designing a system where impurities are retained on the polymeric mesh while target macromolecules flow through unbound. This reverse strategy eliminates the need for elution steps required in conventional capture methods, streamlining the process and improving product recovery efficiency while maintaining reliable separation.
3Manufacturing precision
If low molecular weight impurities are retained in SEC, then purification is achieved, but target macromolecule recovery is limited by low loading capacity
Solution Approach 1:
The patent transitions from traditional column chromatography to a mesh-based separation system that utilizes three-dimensional pore structure. This dimensional approach allows impurities to be retained within the internal pore volume of the mesh while target macromolecules are excluded and pass through. The extensive internal surface area and volume of the polymeric mesh provide high loading capacity without compromising purification quality.
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 allows for high-yield recovery of target proteins by excluding them from the mesh pores while retaining impurities, achieving >90% depletion of host cell proteins and other contaminants in a single step, thereby improving the efficiency and cost-effectiveness of the purification process.
Implementation Method 1
utilizing a cross-linked amino polymer immobilized on a porous support material, enabling efficient depletion of host cell proteins, nucleic acids, and other impurities through steric exclusion and non-covalent interactions
Implementation Method 2
utilizing a cross-linked amino polymer immobilized on a porous support material, enabling efficient depletion of host cell proteins, nucleic acids, and other impurities through steric exclusion and non-covalent interactions
Data Source
AI summary
Method for recovering a target protein from a feedstock comprising said target protein and at least one impurity compound selected from host cell proteins (HCP), DNA, RNA or other nucleic acid, the target protein being characterized by a hydrodynamic radius Rh1 and the impurity compound being characterized by a hydrodynamic radius Rh2, wherein Rh1>Rh2, comprising the following steps (i) to (iv) and optionally step (v): (i) providing a polymeric mesh comprising at least one crosslinked polymer containing positively charged amino groups, wherein the polymer has a pore size exclusion limit Rhi which can be set variably; (ii) adapting the variable pore size exclusion limit Rhi of the polymeric mesh such that Rh2<Rhi and Rh1>Rhi; (iii) contacting the polymeric mesh with the feedstock; (iv) separating the polymeric mesh containing the retained impurity compound from the feedstock containing the excluded target protein.