Transgenic Mouse Producing Human NGF-Beta Protein
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Solution Overview
Problem
Current methods for producing human NGF-β protein using mammal cells result in low yields, high costs, and poor bioactivity, while mouse NGF has limited therapeutic efficacy and immunogenicity, and gene knock-out strategies face limitations in studying NGF-receptor interactions.
Innovation Solution
A method involving homologous recombination to replace the NGF-β gene of mice with the human NGF-β gene, allowing for stable expression and extraction of human NGF-β protein from submandibular glands, which is then formulated into a pharmaceutical composition for therapeutic use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If mammal cells are used to express human NGF-β, then bioactivity is improved, but production cost increases and yield decreases
Solution Approach 1:
The patent extracts the NGF-β gene from human cells and transfers it to mouse submandibular gland cells, which naturally produce high levels of NGF. This allows the host organism (mouse) to serve as a biological factory, extracting human NGF-β protein at high yields while maintaining bioactivity through proper post-translational modifications in the mammalian expression system.
Solution Approach 2:
The patent uses a viral vector (adeno-associated virus or lentivirus) as an intermediary to deliver the human NGF-β gene into mouse submandibular gland cells. This viral mediator enables efficient gene transfer and stable integration, achieving both high expression levels and proper protein folding/modification that preserves bioactivity.
2Productivity
If general bacteria or yeast expression systems are used, then production cost decreases, but bioactivity is reduced due to lack of natural modification and dimerization
Solution Approach 1:
The patent changes the expression system from simple bacterial/yeast systems to a mammalian cell system (mouse submandibular gland), which provides the necessary cellular environment for proper protein folding, disulfide bond formation, and dimerization. This parameter change in the biological system enables natural post-translational modifications that are critical for NGF-β bioactivity.
3Ease of manufacture
If mouse NGF is used, then production is easier and cost is lower, but therapeutic efficacy on human diseases is reduced and immunogenicity increases
Solution Approach 1:
Instead of using mouse NGF for treating human diseases, the patent inverts the approach by putting the human NGF-β gene into mouse cells. This allows production of human NGF-β protein with correct human sequence and structure, maintaining therapeutic efficacy for human diseases while utilizing the easy-to-manage mouse production system.
4Adaptability or versatility
If gene knock-out strategies are used to study NGF-receptor interactions, then research capability is improved, but the ability to study specific interactions is limited
Solution Approach 1:
The patent applies local quality by specifically modifying only the NGF gene in the submandibular gland tissue while leaving the rest of the mouse genome unchanged. This allows targeted overexpression of human NGF-β in a specific tissue, enabling study of NGF-receptor interactions without the need for whole-organism knock-out strategies, thus preserving the ability to study specific molecular interactions.
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 enables large-scale, economically viable production of high bioactivity human NGF-β protein, reducing immunogenicity and improving therapeutic efficacy compared to mouse NGF, while allowing for targeted investigation of NGF-receptor interactions.
Implementation Method 1
using the techniques of homologous recombination, making the mouse thus obtained express human NGF-β gene
Data Source
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AI summary
The present invention provides a transgenic rodent, wherein the genome of said rodent comprising the mutant of NGF-β gene, said rodent expressing the mutein of NGF-β. The present invention also provides the method for obtaining the transgenic rodent, and the methods of preparing the corresponding mutein of mutant NGF-β gene, and the protein obtained using this method. The present invention provides a new way to produce human NGF, and exploit a new way to investigate the function of Mutant NGF-β and its receptor in the level of whole animal, and to screen and purify the mutant NGF-β of high activity and safety.