Mutant Msp Nanopores for Low-Cost Precise Analyte Detection
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Solution Overview
Problem
Current methods for identifying and characterizing analytes, such as nucleic acids and proteins, are laborious and costly, requiring expensive reagents and detection technologies.
Innovation Solution
Development of mutant single-chain Mycobacterium smegmatis porins (Msp) with specific mutations at positions like P97, I68, S73, S116, P123, or V128, which form a tunnel between conductive media to detect analytes by translocation under an electric field, measuring ion current changes for detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current detection methods are used to identify and characterize analytes, then detection can be performed, but the process becomes laborious and costly requiring expensive reagents and detection technologies
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid positions (P97, I68, S73, S116, P123, V128) in the Msp porin protein to optimize its detection capabilities. These parameter changes in the protein structure enable enhanced precision in analyte detection while maintaining a relatively simple system architecture, directly resolving the contradiction between measurement precision and device complexity
Solution Approach 2:
The patent uses a simplified model system based on Msp porin nanopores that replicates the essential detection function without requiring complex commercial instrumentation. This copying approach creates a cost-effective alternative that maintains detection accuracy while eliminating the need for expensive reagents and sophisticated detection technologies
2Measurement precision
If mutant Msp porins with multiple mutations are used to enhance detection precision, then detection accuracy improves, but the manufacturing and characterization process becomes more complex
Solution Approach 1:
The patent segments the mutation strategy into specific, targeted amino acid positions (P97, I68, S73, S116, P123, V128) rather than random mutagenesis. This segmented approach allows for systematic production and characterization of mutants, making the manufacturing process more manageable while achieving enhanced detection precision through focused modifications
Solution Approach 2:
The patent applies local quality by introducing mutations at specific amino acid positions within the Msp porin structure rather than throughout the entire protein. This localized modification strategy enables precise control over protein properties and simplifies manufacturing by focusing changes to specific regions that directly impact detection functionality
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
Provides a cost-effective and efficient method for detecting analytes by enhancing the precision and interaction of nucleic acids and proteins through the Msp tunnel, reducing translocation velocity and increasing detection accuracy.
Implementation Method 1
when the system is subjected to an electric field sufficient to translocate the analyte from one conductive liquid medium to the other
Implementation Method 2
measuring ion current, wherein a reduction in the ion current indicates the presence of the analyte in the first medium
Data Source
AI summary
Provided herein are mutant single-chain Mycobacterium smegmatis porin (Msp) and uses thereof.