Protein Purification From Fermentation Solids by pH-Shift Washing
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Solution Overview
Problem
Existing large-scale protein purification methods from fermentation broths require extensive dilution and complex multi-step procedures, leading to unsatisfactory yields and inability to facilitate a continuous process.
Innovation Solution
A method involving one or more washing steps with a solution that favors solubilization and desorption of proteins from particulate matter by adjusting pH to above or below the protein's pI value and using a positively charged compound to achieve a conductivity of 1-100 mS/cm.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If extensive dilution and complex multi-step procedures are used for protein purification, then purification completeness is improved, but productivity and yield deteriorate
Solution Approach 1:
The patent applies parameter changes by adjusting pH to values above or below the protein's isoelectric point (pI) to alter the charge state of the protein, enabling solubilization from particulate matter. This single parameter change (pH adjustment) replaces complex multi-step procedures including extensive dilution, achieving both high purification completeness and productivity.
Solution Approach 2:
The patent extracts the protein of interest directly from the particulate matter of the fermentation broth through washing steps, separating it from the solid biomass. This extraction approach eliminates the need for extensive dilution and complex multi-step purification procedures, thereby improving both yield and productivity while maintaining purification completeness.
2Stability of the object's composition
If extensive dilution is performed, then protein solubilization is improved, but loss of substance increases
Solution Approach 1:
The patent changes the pH parameter to above or below the protein's pI value, which alters the electrostatic interactions between the protein and particulate matter. This enables effective solubilization and recovery of the protein without requiring extensive dilution, thereby maintaining high protein yield while achieving complete solubilization.
Solution Approach 2:
The patent uses a washing solution as an intermediary medium that facilitates the transfer of protein from particulate matter to solution phase. This washing solution, adjusted to appropriate pH conditions, acts as a mediator that enables protein recovery without requiring large volumes of dilution, thus preventing substance loss.
3Manufacturing precision
If complex multi-step procedures are used, then purification effectiveness is improved, but device complexity and operational difficulty increase
Solution Approach 1:
The patent achieves effective purification through simple parameter changes (pH adjustment) rather than complex multi-step procedures. By adjusting pH to above or below the protein's pI value, the method enables effective separation and purification using straightforward washing steps, thereby reducing device and process complexity while maintaining high purification effectiveness.
Solution Approach 2:
The patent enables a continuous purification process where the washing steps can be performed continuously on the particulate matter. This continuous action approach eliminates the need for complex batch processing steps, reducing operational difficulty and device complexity while maintaining high purification effectiveness throughout the process.
4Stability of the object's composition
If pH is adjusted away from pI value, then protein solubilization is improved, but binding to particulate matter increases
Solution Approach 1:
The patent applies parameter changes by adjusting pH to values above the protein's pI value, which creates a net negative charge on the protein. This charge state reduces hydrophobic interactions and enhances solubilization. The method carefully controls this parameter change to achieve solubilization while minimizing unwanted binding to particulate matter through subsequent washing steps.
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
Enables efficient protein recovery from fermentation broths without extensive dilution, allowing for a continuous purification process and reducing protein loss.
Implementation Method 1
adjusting pH to above or below the protein's pI value
Implementation Method 2
using a positively charged compound to achieve a conductivity of 1-100 mS/cm
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
Implementation Method 4
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.