Feed Media Stabilization Using Pyruvate and pH Adjustment
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Solution Overview
Problem
Concentrated feed media used in mammalian cell cultures, particularly for CHO cells, often precipitate during storage due to components like cysteine and tyrosine, introducing uncertainty and instability, which is undesirable in commercial protein production processes subject to stringent regulatory controls.
Innovation Solution
Incorporating pyruvate into the feed medium at concentrations of 9 to 50 mM, along with cysteine and tyrosine, to stabilize the medium for at least one to three weeks at room temperature, maintaining pH between 5.8 and 7.4, thereby preventing precipitation and ensuring solubility of amino acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If concentrated feed media containing cysteine and tyrosine are used to improve protein production, then the quantity and quality of protein produced increases, but the medium precipitates during storage introducing uncertainty
Solution Approach 1:
The patent changes the pH parameter of the feed medium to a acidic range (pH 2.0-4.5) where cysteine and tyrosine remain soluble. This parameter change resolves the contradiction by maintaining both high concentrations of amino acids for protein production and storage stability without precipitation, as the acidic pH prevents the solubility issues that occur at neutral pH
Solution Approach 2:
The patent performs preliminary adjustment of pH to acidic levels before storage, ensuring that amino acids remain in soluble form throughout the storage period. This preliminary action prevents precipitation from occurring during storage, thereby maintaining reliability while preserving the high productivity benefits of concentrated amino acid formulations
2Productivity
If high concentrations of amino acids are used to improve protein titer, then protein production increases, but the medium becomes unstable and precipitates
Solution Approach 1:
The patent applies parameter change by adjusting pH to the acidic range (pH 2.0-4.5), which fundamentally alters the solubility characteristics of amino acids. At this pH, both cysteine and tyrosine remain soluble even at high concentrations, thereby maintaining medium composition stability while enabling high protein titers through concentrated amino acid formulations
3Loss of time
If feed medium is stored for extended periods to improve operational efficiency, then storage time increases, but precipitation occurs making concentrations unknown
Solution Approach 1:
The patent changes the pH parameter to acidic levels (pH 2.0-4.5) to extend the storage period while maintaining concentration accuracy. The acidic pH creates a stable environment where amino acids do not precipitate over time, allowing the medium to be stored for extended periods with confidence that the original concentrations remain accurate and reliable
Data Source
AI summary
The invention provides stable feed media containing pyruvate and methods for stabilizing feed media by adding pyruvate. The invention further provides methods for producing proteins using such media and proteins produced through the use of such methods.