Lysin-Derived Cell Wall Binding Domain Complexes for Multiplex Bacterial Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Traditional ELISA and PCR methods for bacterial pathogen detection face challenges such as high false positives and negatives, non-specific binding, and high costs due to the need for specifically designed antibodies and species-specific primers, limiting their effectiveness, especially in detecting bacterial pathogens in complex samples.
Innovation Solution
A bacterial pathogen detection system utilizing a lysin-derived cell wall binding domain, combined with a detection domain and a linker, which selectively binds to bacterial cell walls and produces a detectable signal, such as enzymatic or fluorescent activity, to immobilize and identify target pathogens with high specificity, reducing false positives and improving sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional ELISA uses specifically designed antibodies for capturing and detecting bacterial pathogens, then detection capability is improved, but cost and labor increase significantly
Solution Approach 1:
The patent extracts the cell wall binding function from complex antibody molecules and isolates it into a separate, simpler domain (CBD) that can be produced more easily. This domain is then combined with detection domains to create the full detection system, separating the binding function from the detection function to reduce overall system complexity and cost.
Solution Approach 2:
The detection system is segmented into distinct functional modules: cell wall binding domains (CBDs) for pathogen capture, detection domains for signal generation, and linkers for connecting components. This modular approach allows each component to be optimized independently and reduces the overall complexity compared to using single-molecule antibodies for all functions.
2Adaptability or versatility
If traditional ELISA uses conserved epitopes for detection, then cross-species detection is enabled, but false positives and non-specific binding increase
Solution Approach 1:
The patent applies local quality by using lysin-derived CBDs that have evolved to recognize specific local structures in bacterial cell walls (such as peptidoglycan motifs) rather than broad conserved epitopes. This localized recognition provides species-specific or genus-specific binding, improving reliability while maintaining the ability to detect pathogens across related species.
3Measurement precision
If PCR uses species-specific primers for high sensitivity detection, then detection sensitivity is improved, but false positives and negatives increase due to contaminants
Solution Approach 1:
The patent introduces cell wall binding domains as intermediary elements that physically bind to and concentrate bacterial pathogens before detection occurs. This intermediary step acts as a filter that enriches the target pathogen signal while excluding contaminants, thereby improving both sensitivity and reliability by reducing false positives and negatives compared to direct PCR detection.
4Measurement precision
If detection systems use multiple enzymes and fluorescent materials for enhanced signal, then detection sensitivity is improved, but system complexity increases
Solution Approach 1:
The patent merges multiple detection domains with different signal generation capabilities (enzymatic, fluorescent, luminescent) into a single integrated detection complex that can be attached to a single CBD. This combining approach allows enhanced detection sensitivity through signal amplification while managing complexity by consolidating multiple functions into one modular unit rather than requiring separate detection systems for each signal type.
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 system achieves selective and sensitive detection of bacterial pathogens with reduced non-specific binding, enabling reliable and multiplex detection, even in complex samples, with improved limits of detection and reduced false positives, suitable for point-of-care diagnostics.
Implementation Method 1
a lysin-derived cell wall binding domain configured to bind a cell wall of a target bacterial pathogen
Implementation Method 2
the signal for detection is enzymatic activity of the detection domain
Implementation Method 3
the signal for detection is a fluorescent signal
Implementation Method 4
at least one of the lysin-derived cell wall binding domain and the detection domain are biotinylated. In some embodiments, the linker is an avidin linker
Data Source
AI summary
Methods and systems are directed to multiplex detection of a bacterial pathogen in a sample. A first biotinylated lysin-derived cell wall binding domain is complexed with an avidin layer on a surface. A first bacterial pathogen detection complex including a second biotinylated lysin-derived cell wall binding domain, a detection domain, and an avidin linker complexed between the cell wall binding domain and the detection domain is also provided. The cell wall binding domains are derived from an endolysin, autolysin, bacteriocin, or exolysin, and are configured to bind a cell wall of a target bacterial pathogen. The detection domain includes one or more enzymes, fluorescent material, or DNA for emitting a signal for detection. Target bacterial pathogens present in a sample can thus be detected in a sandwich assay exhibiting increased selectivity and reduced limit of detection relative to traditional ELISA.