Beta-Keratin Extraction at Neutral pH With Low Endotoxin
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Solution Overview
Problem
Existing methods for extracting beta-keratin face challenges due to its denser network and rigidity, requiring strong acids or bases, leading to skin irritation and reduced biocompatibility, and high endotoxin levels, making it unsuitable for FDA approval, with a need for solubility in neutral pH solvents and effective cleaning to meet safety standards.
Innovation Solution
A method involving urea and thioglycolic acid extraction at neutral pH, combined with thorough cleaning using detergents and sterilization, results in a beta-keratin extract with low endotoxin levels and solubility in neutral pH solvents, achieving a yield of 10-70% and maintaining thiol groups for stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If strong acids or bases are used to extract beta-keratin, then extraction efficiency is improved, but skin irritation and reduced biocompatibility occur
Solution Approach 1:
The patent changes the chemical parameters of the extraction system by using a neutral pH buffer system (phosphate or carbonate buffer at pH 7-9) combined with urea (2-10% w/v) and thioglycolic acid (0.1-5% w/v), replacing the traditional strong acid or base extraction methods. This parameter change maintains extraction efficiency while eliminating skin irritation and biocompatibility issues.
Solution Approach 2:
The patent introduces intermediary substances (urea and thioglycolic acid) that mediate the extraction process. Urea acts as a chaotropic agent to disrupt protein structure, while thioglycolic acid provides thiol groups that can reduce disulfide bonds. These intermediaries enable effective beta-keratin extraction without requiring strong acids or bases, thus maintaining biocompatibility.
2Ease of manufacture
If strong acids or bases are used to extract beta-keratin, then extraction is facilitated, but protein monomers and oligomers are negatively altered
Solution Approach 1:
The patent changes the extraction conditions to neutral pH (7-9) using phosphate or carbonate buffers, avoiding the extreme pH conditions that cause protein denaturation and degradation. This parameter change facilitates extraction while preserving the structural integrity and functional properties of beta-keratin monomers and oligomers.
Solution Approach 2:
The patent employs urea and thioglycolic acid as intermediary agents that gently disrupt protein structures through chaotropic effects and thiol reduction, respectively. These intermediaries enable extraction without the harsh effects of strong acids or bases, thereby maintaining protein composition stability and preventing unwanted chemical modifications.
3Device complexity
If beta-keratin is extracted without thorough cleaning, then extraction process is simplified, but high endotoxin levels make it unsuitable for FDA approval
Solution Approach 1:
The patent incorporates preliminary cleaning actions into the extraction protocol, including mechanical removal of contaminants, washing steps, and the use of thimerosal (0.01-0.1% w/v) as a preservative to prevent microbial growth. These preliminary actions ensure endotoxin reduction and safety compliance before the actual extraction occurs, making the process suitable for FDA approval.
Solution Approach 2:
The patent introduces thimerosal as an intermediary preservative agent that prevents microbial contamination during extraction. This intermediary substance ensures sterility and prevents endotoxin generation, thereby meeting FDA safety requirements without significantly complicating the overall extraction process when integrated into the protocol.
4Object-affected harmful factors
If beta-keratin extracts are made soluble in neutral pH solvents, then biocompatibility is improved, but extraction difficulty increases due to denser network and rigidity
Solution Approach 1:
The patent uses urea and thioglycolic acid as intermediary agents that facilitate the extraction of beta-keratin from its dense, rigid network structure. Urea disrupts the network through chaotropic effects, while thioglycolic acid reduces disulfide crosslinks. These intermediaries enable extraction that yields neutral pH-soluble, biocompatible beta-keratin without excessive difficulty.
Solution Approach 2:
The patent changes the extraction parameters to include neutral pH buffers (phosphate or carbonate at pH 7-9) combined with specific concentrations of urea (2-10% w/v) and thioglycolic acid (0.1-5% w/v). These parameter changes enable the extraction of beta-keratin while maintaining its solubility in neutral pH solvents and improving biocompatibility, despite the denser network structure.
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
The method produces beta-keratin extracts that are soluble in neutral pH solvents, meet FDA safety standards with low endotoxin levels, and maintain thiol groups, enabling safe and effective use as biomaterials.
Implementation Method 1
A method involves urea and thioglycolic acid extraction at neutral pH
Implementation Method 2
results in a beta-keratin extract with low endotoxin levels and solubility in neutral pH solvents
Implementation Method 3
maintaining thiol groups for stability
Implementation Method 4
combined with thorough cleaning using detergents and sterilization
Implementation Method 5
the resultant beta-keratin extracts are not soluble in neutral and near neutral pH solvents. Instead, beta-keratin extracts often require the use of strong acids or bases
Data Source
AI summary
The present application relates to keratin protein extracts, compositions, methods and uses thereof. In some embodiments, the keratin protein extract is a beta-keratin extract. In some embodiments, the compositions are beta-keratin compositions. In some embodiments, the methods are methods of cleaning a keratin source material, methods of extracting beta-keratin, methods of preparing keratin compositions, and methods of using keratin protein extracts and compositions.