Deep Eutectic Solvent Fixation for Biomolecule Integrity
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Solution Overview
Problem
Current methods for stabilizing biomolecules like RNA, DNA, and proteins during sample storage are inadequate, as they often require specialized reagents and procedures, and existing solutions like formalin fixation can degrade RNA and affect immunohistochemistry, while existing stabilizers are toxic or flammable.
Innovation Solution
The use of deep eutectic solvents (DES) to stabilize and preserve biomolecules by combining components such as Choline chloride and Urea, which form a mixture that effectively fixes cell morphology and inhibits enzymatic degradation, allowing for improved RNA, DNA, and protein integrity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Shape
If formalin fixation is used to preserve cell morphology, then cell structure is maintained, but RNA degradation occurs and immunohistochemistry is affected
Solution Approach 1:
The invention changes the chemical parameters of the fixation medium by using deep eutectic solvents (DES) with specific hydrogen bond donor-acceptor combinations instead of traditional formalin. This parameter change allows simultaneous achievement of cell morphology preservation and RNA integrity maintenance, resolving the contradiction between shape preservation and molecular reliability.
Solution Approach 2:
The invention employs composite deep eutectic solvent systems combining hydrogen bond acceptors (e.g., choline chloride) with hydrogen bond donors (e.g., urea, formamide). These composite DES formulations provide multiple protective functions simultaneously - fixing cell structure while preventing RNA degradation, thus resolving the contradiction through material composition rather than single-agent action.
2Reliability
If traditional stabilizing reagents are used to prevent biomolecule degradation, then biomolecule integrity is improved, but toxicity and environmental harm increase
Solution Approach 1:
The invention changes the chemical composition parameters from toxic traditional reagents (formalin, phenol, chloroform) to non-toxic deep eutectic solvents based on natural components like choline chloride, urea, and formamide. This parameter change in chemical composition maintains biomolecule integrity while eliminating toxicity, resolving the contradiction between reliability and harmful factors.
Solution Approach 2:
The invention uses readily available, biodegradable components (choline chloride, urea, formamide) that can be easily disposed of without environmental harm. These replace expensive, persistent, and toxic reagents, achieving both biomolecule protection and environmental friendliness.
3Reliability
If multiple specialized reagents are used to stabilize different biomolecules, then stabilization effectiveness is improved, but procedure complexity increases
Solution Approach 1:
The invention creates a universal deep eutectic solvent system that simultaneously stabilizes multiple biomolecule types (RNA, DNA, proteins) and preserves cell morphology. This single multi-functional DES replacement eliminates the need for multiple specialized reagents and complex procedural steps, resolving the contradiction between stabilization effectiveness and procedure complexity.
Solution Approach 2:
The invention merges multiple stabilization functions (RNA protection, DNA protection, protein stabilization, cell fixation) into a single deep eutectic solvent system. By combining these functions in one reagent rather than using separate agents, the procedure complexity is reduced while maintaining comprehensive stabilization effectiveness.
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
DES mixtures effectively stabilize RNA, DNA, and proteins, maintaining cell morphology and preventing degradation, even in challenging storage conditions, while being non-toxic and environmentally friendly, thus enhancing the quality and integrity of biomolecules for diagnostic and analytical purposes.
Implementation Method 1
The deep eutectic solvent comprises a hydrogen bond acceptor and a hydrogen bond donor
Implementation Method 2
effectively fixes cell morphology and inhibits enzymatic degradation
Data Source
Figure 1A~2
Figure 3~4
Figure 5A~5B
AI summary
Provided is a use of a deep eutectic solvent to inhibit the degradation of a biomolecule, wherein the biomolecule is a protein. The deep eutectic solvent comprises a first component and a second component. The first component is a compound of Formula I: wherein: R6 is H or OH; R7 is selected from H, CH3, Cl, Br, a carbonyl oxygen, and Z is selected from -CH2-, O and S; and R8 is R11 or OH. The second component comprises a compound of Formula II or a salt thereof: wherein: A is selected from O, S, and NH; R1 is selected from R9 and -C(R3)(R4)(R5); R2 is selected from H and linear alkyl group having 1 to 3 carbon atoms; R3 is an optionally substituted 5- or 6-membered aliphatic or aromatic ring, wherein the substituent is R10; R4 and R5 are F. R9 is a mono-, di- or trifluoromethyl group or a mono-, di- or tri-fluoroethyl group; and R10 and R11 are each independently selected from alkyl groups having one to three carbon atoms, monochloroalkyl groups having one to three carbon atoms, and mono-, di- or tri-fluoroalkyl groups having one to three carbon atoms.