Affinity Membrane Preparation for High-Capacity Biologics Purification
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Solution Overview
Problem
Existing resin-based chromatography methods for biologics purification face challenges with low binding capacity at short residence times, leading to low productivity and potential product degradation, especially for larger biologics, and there is a lack of affinity membrane chromatography products suitable for high-throughput purification.
Innovation Solution
A method for preparing affinity membranes by immersing a membrane in swelling solvents with coupling reagents and adsorptive groups, followed by incubation with biologic-target-specific solutions, resulting in high binding capacity and short residence times, using macroporous supports like regenerated cellulose membranes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If resin-based chromatography is used to achieve high binding capacity, then binding capacity is improved, but residence time increases leading to low productivity
Solution Approach 1:
The patent uses macroporous hydrogel beads with large pore sizes (1-10 μm) to enable rapid mass transfer of biologics while maintaining high binding capacity. The macroporous structure allows proteins to quickly access the interior surface where ligands are immobilized, achieving high productivity without sacrificing binding capacity.
Solution Approach 2:
The patent immobilizes ligands specifically on the interior surface of the macroporous hydrogel beads, creating a localized high-density ligand region. This concentrates the binding function in the optimal location where mass transfer is fastest, maximizing both binding capacity and residence time efficiency.
2Quantity of substance
If small pore structures are used in resin-based columns, then binding capacity is improved, but mass transfer becomes slow leading to long residence times
Solution Approach 1:
The patent employs macroporous hydrogel beads with pore sizes of 1-10 μm, which are significantly larger than conventional resin pores. This macroporous structure enables rapid diffusion and convection of biologics through the bead interior, achieving fast mass transfer rates while maintaining high binding capacity through ligand immobilization on the interior surface.
3Quantity of substance
If conventional resin chromatography products are used, then high binding capacity is achieved at long residence times, but no products are available for short residence times
Solution Approach 1:
The patent uses macroporous hydrogel beads with 1-10 μm pores that enable rapid mass transfer, allowing the system to achieve high binding capacity at residence times of seconds rather than minutes. This macroporous structure is the key enabler for short residence time operation while maintaining productivity.
Solution Approach 2:
The patent changes the pore size parameter from conventional small pores to macropores (1-10 μm), which fundamentally alters the mass transfer regime from diffusion-limited to convection-enhanced transport. This parameter change enables the system to operate effectively at short residence times while maintaining high binding capacity.
4Adaptability or versatility
If larger biologics are purified using resin-based columns, then purification is possible, but binding capacity decreases and columns are prone to clogging
Solution Approach 1:
The patent uses macroporous hydrogel beads with large pore sizes (1-10 μm) that can accommodate larger biologics such as virus particles, virus-like particles, and large proteins without steric hindrance. The large pores prevent clogging while maintaining high binding capacity through ligand immobilization on the interior surface.
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
The method achieves static protein binding capacities of 60-100 mg/mL and dynamic binding capacities of 20-90 mg/mL at 6 seconds or less with low backpressure, significantly enhancing productivity and overcoming limitations of resin-based systems.
Implementation Method 1
a membrane for purifying biologics such as proteins, polypeptides, peptides, polynucleotides, nucleotides, viral vectors, and vaccines using affinity separation methods
Implementation Method 2
immersing a membrane into a first solution of a coupling reagent in a first swelling solvent solution to swell said membrane and increase exposure of reactive sites on said membrane
Data Source
AI summary
A method for preparing an adsorptive media for binding biologic molecules comprising immersing a macroporous support in a first solution of a coupling reagent in a solvent solution for attachment of said coupling reagent to form coupling groups; and, immersing said macroporous support in an incubating solution selected from the group consisting of ligand, nucleotide, oligonucleotide, peptide, polypeptide, protein, and enzyme solutions having an affinity to a biologic target molecule to couple one of said ligands, nucleotides, oligonucleotides, peptides, polypeptides, proteins, and enzymes to at least a portion of said coupling groups of said macroporous support for binding with said biologic target molecule when exposed to said macroporous support.