Polysorbate Analysis After Protein Precipitation in High-Protein Formulations
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Solution Overview
Problem
Existing methods for analyzing polysorbate concentration in protein drug formulations are hindered by high sample viscosity, gel formation, column clogging, and interfering peaks, especially in samples with high protein concentrations, leading to inaccurate results.
Innovation Solution
A method involving the addition of a chelating agent, such as EDTA, and a C1-C6 alcohol, like ethanol, to precipitate proteins from the formulation, followed by centrifugation to separate the protein and allow accurate polysorbate analysis using mixed-mode anion exchange-hydrophobic high-performance liquid chromatography.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If existing methods for analyzing polysorbate concentration are used in high protein samples, then polysorbate analysis can be performed, but the results become inaccurate due to high sample viscosity, gel formation, column clogging, and interfering peaks
Solution Approach 1:
The patent extracts and removes proteins from the sample using precipitation methods (adding organic solvents like ethanol or acetone, or using chelating agents) before polysorbate analysis. This separation eliminates the harmful effects of high protein concentration on viscosity, gel formation, and column clogging, while preserving polysorbate for accurate measurement
Solution Approach 2:
The patent performs preliminary protein precipitation and removal steps before the actual polysorbate analysis. This preliminary action prepares the sample by eliminating interfering substances, ensuring that the subsequent polysorbate measurement is not affected by viscosity, gel formation, or column clogging issues
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 removes proteins from the sample, reducing interference and enabling accurate polysorbate quantification, even in high-protein formulations, with improved specificity and accuracy, suitable for quality control environments.
Implementation Method 1
adding to the polysorbate-containing composition that comprises a protein and a polysorbate, an amount of a chelating agent and an amount of a C1-C6 alcohol that together are effective to precipitate the protein
Implementation Method 2
allow accurate polysorbate analysis using mixed-mode anion exchange-hydrophobic high-performance liquid chromatography
Data Source
AI summary
Provided herein are methods of precipitating proteins within compositions that contain a protein and a polysorbate. These methods allow for an accurate determination of the concentration or the amount of polysorbate within the composition.