MIS Storage Buffer Composition for Proteolysis Control
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Solution Overview
Problem
Purified Müllerian Inhibiting Substance (MIS) is prone to cleavage and degradation by proteolytic enzymes, leading to decreased shelf-life and loss of bioactivity.
Innovation Solution
A storage buffer composition comprising deionized water, buffering agent (e.g., HEPES), salt (NaCl), sugar alcohol (mannitol), gelatin, surfactant (Tween-20), and biocide (Micr-O-protect) stabilizes MIS by preventing proteolysis, maintaining bioactivity at 2°C to 8°C.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If purified MIS is stored without stabilization measures, then the storage buffer is simple, but the shelf-life is short and bioactivity is lost due to proteolytic degradation
Solution Approach 1:
The storage buffer employs a composite formulation containing multiple stabilizing components including sugar alcohols (mannitol, sorbitol), gelatin, surfactants (Tween-20), and biocides (Micr-O-protect). This composite approach addresses proteolytic degradation through synergistic action of different ingredients, extending MIS shelf-life while maintaining bioactivity without requiring overly complex processing or storage conditions
Solution Approach 2:
Gelatin acts as an intermediary protective agent in the storage buffer, forming a protective matrix around the MIS protein that physically shields it from proteolytic enzymes. This mediator approach allows the MIS to remain stable in storage without requiring complete elimination of proteases, thereby extending shelf-life while keeping the buffer composition manageable
2Reliability
If conventional storage buffers are used, then the buffer composition is simple, but proteolytic degradation occurs leading to loss of bioactivity
Solution Approach 1:
The storage buffer converts the potentially harmful presence of proteolytic enzymes into a beneficial situation by adding biocides (Micr-O-protect) that specifically target and neutralize these enzymes. Additionally, the surfactant Tween-20 is used to disrupt enzyme-protein interactions, transforming the degradation threat into a stable storage condition while maintaining a relatively simple buffer formulation
Solution Approach 2:
The storage buffer optimizes physical-chemical parameters including pH (maintained by buffering agents), ionic strength (adjusted with salts like NaCl), and osmolarity (controlled by sugar alcohols like mannitol). These parameter optimizations create an environment that inherently stabilizes MIS protein structure and reduces susceptibility to proteolytic degradation, enhancing reliability without excessive complexity
3Duration of action of stationary object
If MIS is stored at extended time periods, then long-term storage is achieved, but concentration loss increases beyond acceptable levels
Solution Approach 1:
The storage buffer performs preliminary protective actions by pre-incorporating stabilizing agents that prevent degradation before it occurs. Sugar alcohols like mannitol and sorbitol are included to maintain proper hydration and prevent protein aggregation, while gelatin provides a protective matrix. This preliminary stabilization allows MIS to be stored for extended periods with concentration loss limited to 10% or less over 60 days
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 buffer extends MIS shelf-life and maintains bioactivity, with concentration loss reduced to 10% or less over 60 days, suitable for diagnostic use.
Implementation Method 1
The at least one sugar alcohol is mannitol... stabilizes MIS by preventing proteolysis
Implementation Method 2
The at least one sugar alcohol is mannitol... stabilizes MIS by preventing proteolysis
Implementation Method 3
gelatin... stabilizes MIS by preventing proteolysis
Implementation Method 4
surfactant (Tween-20)... stabilizes MIS by preventing proteolysis
Implementation Method 5
buffering agent (e.g., HEPES)... stabilizes MIS by preventing proteolysis
Implementation Method 6
biocide (Micr-O-protect) stabilizes MIS by preventing proteolysis
Data Source
Figure 1A~1B
Figure 1C~1D
Figure 1E
AI summary
Non-limiting embodiments of an improved Müllerian Inhibiting Substance storage buffer composition(s) and method(s) of stabilizing a Müllerian Inhibiting Substance in the improved storage buffer composition.