Thiol-ene Click Chemistry for Protein Modification
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Solution Overview
Problem
Current methods for covalent modification of proteins are limited by stability, solubility issues in organic solvents, and require harsh conditions, leading to inefficient and pH-dependent reactions with potential protein damage and undesirable side reactions.
Innovation Solution
The use of thiol-ene and thiol-yne chemistries with click-like properties, such as norbornene and vinyl ether moieties, for rapid, pH-independent covalent modification of proteins under non-denaturing conditions, allowing for efficient and selective crosslinking without self-reactivity and protein damage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional covalent modification methods are used in aqueous environment, then proteins can be modified, but the reaction is pH-dependent and requires harsh conditions leading to protein damage
Solution Approach 1:
The patent changes the chemical parameters of the reaction system by introducing thiol-ene click chemistry that operates under mild, physiological conditions rather than harsh pH-dependent conditions. This allows efficient protein modification without denaturation or damage to the protein structure.
Solution Approach 2:
The patent uses thiol groups as intermediary functional groups that enable selective and mild crosslinking. The thiol-ene click chemistry mechanism provides a gentle pathway for covalent modification that avoids the harsh conditions typically required for conventional protein crosslinking methods.
2Reliability
If conventional crosslinking chemistry is used, then proteins can be crosslinked, but self-reaction and side reactions occur reducing selectivity
Solution Approach 1:
The patent converts the potential harm of unselective crosslinking into a benefit by using thiol-ene click chemistry, which is inherently selective due to the high reactivity of thiols with ene groups. This selectivity prevents self-reaction and side reactions while enabling efficient crosslinking.
Solution Approach 2:
The patent introduces specific functional groups (thiol and ene groups) at localized positions on the protein molecules. This localized functionalization ensures that crosslinking occurs only at desired sites with high selectivity, preventing unwanted self-reactions and side reactions elsewhere in the system.
3Productivity
If rapid crosslinking is achieved, then modification efficiency increases, but reaction conditions become harsher causing more protein damage
Solution Approach 1:
The patent changes the kinetic parameters of the reaction by employing thiol-ene click chemistry, which proceeds rapidly under mild physiological conditions. This eliminates the traditional trade-off between speed and gentleness, allowing fast crosslinking without protein damage.
4Quantity of substance
If pH optimization is performed to improve solubility, then protein solubility increases, but reactivity of amino acid residues decreases
Solution Approach 1:
The patent introduces thiol groups as intermediary functional groups that provide high reactivity independent of pH. These thiol groups serve as excellent nucleophiles that can react efficiently with ene groups across a broad pH range, eliminating the need to compromise between solubility optimization and reactivity requirements.
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 rapid, efficient, and broadly applicable covalent modification of proteins, reducing side reactions and protein damage, and facilitating the formation of three-dimensional polymer networks suitable for biomedical applications like tissue regeneration.
Implementation Method 1
subjecting a reaction mixture comprising: (a) a biological macromolecule comprising one or more thiol groups; and (b) a molecule comprising one or more olefin or alkyne moieties to a radical reaction under conditions sufficient to produce the covalently modified biological macromolecule
Implementation Method 2
The use of thiol-ene and thiol-yne chemistries with click-like properties, such as norbornene and vinyl ether moieties, for rapid, pH-independent covalent modification of proteins
Implementation Method 3
In another embodiment, the radical reaction is photoinitiated
Data Source
AI summary
The present disclosure provides a method of covalently modifying a biological macromolecule, the method comprising subjecting a reaction mixture comprising: (a) a biological macromolecule comprising one or more thiol groups; and (b) a molecule comprising one or more olefin or alkyne moieties to a radical reaction under conditions sufficient to produce the covalently modified biological macromolecule. The present disclosure also provides a method of covalently modifying a biological macromolecule, the method comprising subjecting a reaction mixture comprising: (a) a molecule comprising one or more thiol groups; and (b) a biological macromolecule comprising one or more olefin or alkyne moieties to a radical reaction under conditions sufficient to produce the covalently modified biological macromolecule. The present disclosure further provides a covalently modified biological macromolecule prepared by any of the disclosed methods. The covalently modified biological macromolecules may be further crosslinked to form a scaffold.