Charged Depth Filtration to Re-Oxidize Antigen-Binding Proteins
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Therapeutic antigen-binding proteins, such as antibodies, are prone to structural instability due to reduced disulfide bonds during production and purification, leading to fragmentation and aggregation, which impair their biological functions and therapeutic efficacy.
Innovation Solution
A method involving charged depth filtration is used to contact antigen-binding protein solutions, promoting re-oxidation and reducing the percentage of reduced protein molecules by at least 20% to 40%, as measured by non-reducing capillary electrophoresis with sodium dodecyl sulfate (nrCE-SDS).
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If standard filtration and chromatography steps are used during production and purification, then antigen-binding proteins can be recovered, but disulfide bonds are reduced leading to structural instability and fragmentation
Solution Approach 1:
The patent applies preliminary action by incorporating a charged depth filtration step early in the purification process, before subsequent chromatography steps. This preliminary filtration removes reducing agents and promotes re-oxidation of disulfide bonds in advance, preventing structural instability and fragmentation that would otherwise occur during later processing steps.
2Ease of manufacture
If reduced antigen-binding protein molecules are allowed to remain during purification, then the purification process can proceed, but the molecules may fragment during storage or clinical use
Solution Approach 1:
The charged depth filtration is applied as a preliminary step that promotes re-oxidation of reduced protein molecules before they enter storage or clinical use. This advance action ensures structural integrity is restored prior to long-term storage, preventing fragmentation during the storage period while maintaining process simplicity.
Solution Approach 2:
The charged depth filter acts as an intermediary medium that facilitates the re-oxidation of reduced antigen-binding protein molecules. The filter's charged surface promotes oxidation reactions, serving as a mediator between the reduced protein molecules and the oxidizing environment, thereby restoring disulfide bonds without requiring complex additional reagents or steps.
3Ease of operation
If disulfide bonds are reduced during production, then protein molecules can be processed, but structural integrity is weakened and therapeutic efficacy is impaired
Solution Approach 1:
The charged depth filter serves as an intermediary that promotes re-oxidation of reduced disulfide bonds during the processing workflow. This intermediary action restores structural integrity to protein molecules that were reduced during production, maintaining both processing capability and structural strength throughout the manufacturing process.
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 effectively stabilizes antigen-binding proteins by enhancing re-oxidation, minimizing fragmentation and aggregation, resulting in a more stable and effective pharmaceutical formulation.
Implementation Method 1
contacting an aqueous solution comprising antigen-binding protein molecules with a charged depth filter under conditions sufficient to enhance re-oxidation of the antigen-binding protein molecules
Data Source
AI summary
Methods of producing an aqueous formulation of an antigen-binding protein or enhancing re-oxidation of an antigen-binding protein are disclosed. The methods comprise (a) contacting an aqueous solution comprising antigen-binding protein molecules with a charged depth filter under conditions sufficient to enhance re-oxidation of the antigen-binding protein molecules and achieve a decrease in the percentage of reduced antigen-binding protein molecules, compared to the percentage of reduced anti-gen-binding protein molecules observed prior to step (a); and (b) optionally, measuring the amount or relative amount of reduced antigen-binding protein molecules. Formulations comprising a re-oxidized antigen-binding protein are also described.