hIGF-I Production via pH Adjustment and Standing Precipitation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing human insulin-like growth factor I (hIGF-I) face challenges in achieving high purity and yield due to the presence of intrinsic proteins and modified hIGF-I (mhIGF-I) isomers, which require complex purification steps and are costly for large-scale production.
Innovation Solution
A method involving the use of recombinant coryneform bacteria to produce hIGF-I, where the culture liquid pH is adjusted to 8 or more and left to stand, followed by removal of bacterial cells, effectively reducing the amount of mhIGF-I and increasing the yield of hIGF-I, thereby simplifying purification and reducing costs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If recombinant E. coli or yeast is used to produce hIGF-I, then hIGF-I can be produced, but intrinsic proteins and misfolded hIGF-I isomers are also produced requiring complex purification
Solution Approach 1:
The patent changes the physical-chemical parameters of the culture liquid by adjusting pH to alkaline conditions (pH 8 or more) and allowing the culture liquid to stand, which causes misfolded hIGF-I isomers to precipitate. This parameter change simplifies purification by enabling simple filtration or centrifugation instead of complex chromatography steps.
Solution Approach 2:
The patent extracts or removes the harmful misfolded hIGF-I isomers from the culture liquid through precipitation and simple filtration/centrifugation steps. By separating the misfolded isomers as a distinct phase that can be easily removed, the purification process becomes much simpler while maintaining high hIGF-I yield.
2Manufacturing precision
If complex chromatography purification steps are used to remove misfolded hIGF-I, then high purity hIGF-I is obtained, but production cost increases
Solution Approach 1:
The patent utilizes pH parameter change to alkaline conditions followed by standing time to induce precipitation of misfolded hIGF-I isomers. This simple parameter-based separation achieves high purity without requiring expensive chromatography columns, solvents, or complex operating procedures, thereby significantly reducing production costs.
Solution Approach 2:
The patent replaces expensive, complex chromatography purification systems with simple, disposable filtration or centrifugation steps. The misfolded isomers are removed through basic filtration membranes or centrifugal separation, which are much cheaper and easier to implement than chromatography systems, making large-scale production economically viable.
3Manufacturing precision
If pH is adjusted to 8 or more and culture liquid is left standing, then misfolded hIGF-I is reduced, but production time increases
Solution Approach 1:
The patent performs pH adjustment to alkaline conditions and allows the culture liquid to stand before proceeding to filtration or centrifugation. This preliminary action causes misfolded hIGF-I isomers to precipitate in advance, so that subsequent separation steps are quick and simple. The time investment is made upfront rather than during complex purification, overall streamlining the process.
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 method enables the production of high-purity hIGF-I with increased yield by minimizing the amount of mhIGF-I, eliminating the need for complex chromatography purification steps and achieving high-purity hIGF-I production at a lower cost.
Implementation Method 1
adjusting the pH of a culture liquid of a human insulin-like growth factor I producing bacteria to 8 or more after completion of culture; letting the culture liquid obtained in step (A) stand
Data Source
Figure 1
AI summary
A method is provided for producing hIGF-I with high purity and yield. This is a method for producing human insulin-like growth factor I, having a step of removing modified human insulin-like growth factor I from the human insulin-like growth factor I, the step including: (A) a step of adjusting the pH of a culture liquid of a human insulin-like growth factor I producing bacteria to 8 or more after completion of culture; (B) a step of letting the culture liquid obtained in step (A) stand; and (C) a step of removing the producing bacteria from the culture liquid obtained in step (B).