Protein Composition for Radiosterilization Without Denaturation
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Solution Overview
Problem
Existing methods for radiosterilizing proteins, such as those involving cooling conditions, complete water removal, or using radical scavengers, are ineffective or costly, and result in protein degradation and denaturation, especially for highly hydrophilic proteins.
Innovation Solution
A method for producing a protein composition that includes a protein, a radical scavenger, and a hydrogen-bond-formable compound, with specific functional groups and ratios, to minimize protein denaturation during radiosterilization by facilitating radical scavenging through hydrogen bonds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If eukaryotic cells are used for producing recombinant proteins, then protein production capacity and post-translational modification capability are improved, but cell death occurs during culture leading to protein degradation and reduced productivity
Solution Approach 1:
The patent applies preliminary action by pre-coating the culture vessel surface with extracellular matrix components (such as fibronectin, laminin, or collagen) before cell culture. This preliminary surface modification creates a protective environment that prevents cell death during culture, thereby maintaining protein production capacity and avoiding protein degradation while improving overall productivity
Solution Approach 2:
The patent uses extracellular matrix components as an intermediary substance between the culture vessel surface and the eukaryotic cells. This intermediary layer (coated on the vessel surface) mediates cell adhesion and protection, preventing direct harmful interactions with the vessel surface and maintaining cell viability throughout the culture process, thus resolving the contradiction between production capacity and productivity
2Reliability
If eukaryotic cells are used for producing recombinant proteins, then post-translational modification capability is improved, but cell death during culture causes protein degradation
Solution Approach 1:
The culture vessel surface is pre-coated with extracellular matrix components before cell culture begins. This preliminary action creates a stable, cell-friendly environment that prevents cell death throughout the culture process, thereby maintaining both post-translational modification capability and protein integrity without degradation
Solution Approach 2:
The extracellular matrix coating acts as a protective cushioning layer that anticipates and prevents cell death during culture. By providing this protective environment in advance, the system ensures cell survival and maintains protein integrity throughout the culture process, resolving the contradiction between modification capability and protein stability
3Ease of operation
If conventional cell culture methods are used, then simplicity of operation is maintained, but cell death occurs leading to protein degradation and reduced yield
Solution Approach 1:
The method requires only a simple pre-coating step of the culture vessel surface with extracellular matrix components before adding cells. This preliminary action is straightforward to perform and maintains the simplicity of operation while dramatically improving cell survival and protein yield, avoiding the complexity of other cell line development approaches
Solution Approach 2:
The extracellular matrix coating serves as a simple intermediary that can be easily introduced into conventional culture protocols. This intermediary layer prevents cell death without requiring complex procedural changes, maintaining ease of operation while significantly reducing protein loss and improving overall yield
Data Source
AI summary
The present invention aims to provide a method for producing a protein composition, the method being capable of suppressing changes such as degradation and denaturation of a protein which occur when a highly hydrophilic protein or a protein that undergoes denaturation by complete removal of water is irradiated with radiation. The present invention provides a method for producing a protein composition containing a protein (A), a radical scavenger (RS), and at least one hydrogen-bond-formable compound (HC) selected from the group consisting of amino acids, peptides, and proteins other than the protein (A), the method including a sterilization step of radiosterilizing an unsterilized protein composition, wherein the unsterilized protein composition contains the protein (A), the radical scavenger (RS), and the hydrogen-bond-formable compound (HC), the protein (A) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the hydrogen-bond-formable compound (HC) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the at least one functional group in the protein (A) is capable of binding to the at least one functional group in the hydrogen-bond-formable compound (HC) via a hydrogen bond, the unsterilized protein composition has a water content of 0 to 30% by weight based on the weight of the unsterilized protein composition, a weight ratio of the radical scavenger (RS) to the protein (A) in the unsterilized protein composition [radical scavenger (RS)/protein (A)] is 0.01 to 1.0, and a molar ratio of a total molar number of functional groups in the hydrogen-bond-formable compound (HC) to a total molar number of functional groups in the protein (A) in the unsterilized protein composition [the total molar number of functional groups in the hydrogen-bond-formable compound (HC)/the total molar number of functional groups in the protein (A)] is 0.01 to 0.50.