Protein Purification From Fermentation Solids Using Desorbing Washes
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Solution Overview
Problem
Existing methods for purifying proteins from fermentation broths require extensive dilution and multi-step procedures, leading to unsatisfactory yields and complications in large-scale protein recovery due to binding and crystallization, preventing a continuous purification process.
Innovation Solution
A method involving one or more washing steps with a washing solution that favors solubilization and desorption of proteins from particulate matter using pH conditions above or below the protein's pI value, optionally with a positively charged compound to achieve a conductivity of 1-100 mS/cm, allowing for continuous purification without significant dilution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If extensive dilution and multi-step procedures are used for protein purification, then purification can be achieved, but yield is reduced and process complexity increases
Solution Approach 1:
The patent applies parameter changes by adjusting pH to values above or below the protein's isoelectric point (pI) during the washing step. This causes the protein to carry a net charge, preventing adsorption to particulate matter and enabling efficient recovery without extensive dilution or multi-step procedures, thereby maintaining both purification quality and high protein yield
Solution Approach 2:
The patent implements a continuous purification process where the washing step with optimized pH conditions continuously recovers protein from particulate matter without interruption. This eliminates the need for batch-wise extensive dilution and multiple sequential purification steps, thereby maintaining high productivity and protein yield while ensuring consistent purification quality
2Stability of the object's composition
If extensive dilution is performed, then protein solubility is improved, but process complexity and time consumption increase
Solution Approach 1:
The patent changes the pH parameter to values above or below the protein's pI during washing, which fundamentally alters protein solubility and charge characteristics. This eliminates the need for extensive dilution to achieve solubility, thereby reducing purification time and process complexity while maintaining protein stability and solubility through controlled charge states
Solution Approach 2:
The patent performs preliminary pH adjustment before the washing step to ensure the protein is in a charged state that prevents adsorption. This preliminary action eliminates the need for subsequent extensive dilution steps that would otherwise be required to maintain solubility, thereby reducing overall purification time and complexity
3Reliability
If pH adjustment is performed, then protein desorption from particulate matter is improved, but process complexity increases
Solution Approach 1:
The patent uses pH adjustment as a simple yet effective parameter change during the washing step to control protein charge and prevent adsorption to particulate matter. This single parameter adjustment achieves reliable protein recovery without requiring complex multi-step procedures, specialized equipment, or additional chemicals, thereby maintaining low process complexity while ensuring high recovery reliability
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
This approach enhances protein recovery by reducing loss and enabling a continuous purification process, improving yield and efficiency by minimizing dilution and simplifying the purification steps.
Implementation Method 1
contacting the particulate matter with a washing solution comprising one or more conditions that favor the solubilization of the protein of interest
Implementation Method 2
a pH value above the pI value of the protein of interest or b) a pH value below the pI value of the protein of interest and under these conditions a positively charged compound
Implementation Method 3
conditions that favor the solubilization of the protein of interest and/or the desorption of the protein of interest from the particulate matter
Data Source
AI summary
The present invention is directed to a method of purifying a protein of interest from the particulate matter of a fermentation broth comprising the step of purifying the protein of interest from the particulate matter of the fermentation broth, wherein the step of purifying the protein of interest from the particulate matter of the fermentation broth comprises one or more washing steps, which comprises contacting the particulate matter with a washing solution comprising one or more conditions that favor the solubilization of the protein of interest and/or the desorption of the protein of interest from the particulate matter.