Esterase Removal in Protein Formulations to Reduce Subvisible Particles
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Solution Overview
Problem
Biopharmaceutical formulations containing proteins are prone to the formation of subvisible particles due to protein aggregation, which is exacerbated by the degradation of surfactants like polysorbates by host cell esterases, leading to stability issues and regulatory compliance challenges.
Innovation Solution
Incorporating a fatty acid ester surfactant, such as polyoxyethylene sorbitan esters, and reducing esterase activity through processes like hydrophobic interaction chromatography to maintain the integrity of the surfactant and prevent particle formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If polysorbate surfactant is added to prevent protein aggregation, then protein stability is improved, but esterase degradation of polysorbate leads to loss of surfactant activity and subvisible particle formation
Solution Approach 1:
The patent changes the chemical parameter of the surfactant from polysorbate (fatty acid ester) to a non-ester surfactant alternative, eliminating the ester bond that is susceptible to esterase degradation. This parameter change resolves the contradiction by maintaining surfactant functionality while removing the degradation pathway that leads to particle formation.
Solution Approach 2:
The patent removes the need for continuous surfactant replacement by selecting a stable surfactant that does not degrade over time. This eliminates the cycle of surfactant degradation and particle formation, providing long-term stability without requiring frequent intervention.
2Object-generated harmful factors
If host cell protein removal is increased to reduce esterase activity, then surfactant degradation is reduced, but formulation complexity and processing steps increase
Solution Approach 1:
The patent extracts the problematic esterase enzyme from the formulation by implementing enhanced purification steps that specifically remove host cell proteins containing esterase activity. This extraction eliminates the source of surfactant degradation while maintaining product quality.
Solution Approach 2:
The patent performs esterase removal as a preliminary purification step before final formulation. By removing the esterase early in the process, subsequent storage and handling do not require additional protective measures, simplifying the overall process.
3Productivity
If protein concentration is increased to improve formulation efficiency, then dosing volume is reduced, but aggregation propensity and subvisible particle formation increase
Solution Approach 1:
The patent introduces a stable surfactant as an intermediary substance that mediates between high protein concentration and aggregation prevention. The surfactant acts as a protective layer around protein molecules, enabling high concentrations to be maintained without increased aggregation risk.
Solution Approach 2:
The patent changes the physical parameter of protein concentration to higher levels while simultaneously changing the chemical environment by adding stable surfactant. This combined parameter change enables improved formulation efficiency without the usual penalty of increased aggregation.
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 composition achieves reduced subvisible particle formation, maintaining stability and compliance with regulatory limits over extended storage periods, ensuring the efficacy and safety of biopharmaceutical formulations.
Implementation Method 1
Incorporating a fatty acid ester surfactant, such as polyoxyethylene sorbitan esters, and reducing esterase activity through processes like hydrophobic interaction chromatography to maintain the integrity of the surfactant and prevent particle formation.
Implementation Method 2
When a polysorbate degrades (hydrolyzes) into its component head group and fatty acid tail, it loses its effectiveness as a protein stabilizer
Implementation Method 3
reducing esterase activity through processes like hydrophobic interaction chromatography
Implementation Method 4
reducing esterase activity through processes like hydrophobic interaction chromatography
Data Source
AI summary
The present disclosure provides a stable protein composition containing a surfactant and having less than 400 subvisible particles of 10 microns or greater diameter per container, or less than 10,000 subvisible particles of 2 microns or greater per container. A method of manufacturing such a stable protein composition is disclosed, which includes a unit of operation that removes or decreases an esterase activity that degrades the surfactant. The unit of operation may be hydrophobic interaction chromatography or filtration, mixed mode chromatography, or the like.