Fluorescent Protein Sensors for Siderophore Detection
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Solution Overview
Problem
Current methods for detecting and monitoring microbes, particularly pathogenic bacteria, are limited in their ability to efficiently detect siderophores and other small molecule compounds secreted by these organisms, which are indicative of microbial presence and pathogenicity.
Innovation Solution
Development of high affinity protein-based sensors that are fluorescently labeled, allowing for the detection, discrimination, and quantification of apo- and ferric siderophores, as well as other microbe-associated compounds, through changes in fluorescence signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used for siderophores and microbe-associated compounds, then the detection process is simpler, but the detection sensitivity and precision are insufficient
Solution Approach 1:
The patent uses fluorescently labeled binding proteins as intermediary sensors that specifically bind to siderophores and microbe-associated compounds. These binding proteins act as mediators between the target compounds and the detection system, enabling sensitive detection through fluorescence signal changes when the binding proteins bind to their target ligands
Solution Approach 2:
The patent employs fluorescent labels that undergo parameter changes (fluorescence intensity or wavelength shifts) upon binding to target compounds. This allows detection sensitivity to be improved by monitoring these optical parameter changes, which occur when the fluorescently labeled binding proteins interact with siderophores or other microbe-associated compounds
2Measurement precision
If high affinity binding proteins are engineered with fluorescent labels for specific detection, then detection precision improves, but the manufacturing complexity increases
Solution Approach 1:
The patent uses recombinant DNA technology to produce multiple copies of engineered binding proteins with fluorescent labels through bacterial expression systems. This allows complex fluorescent sensors to be manufactured efficiently by copying the genetic information and expressing the proteins in large quantities, reducing the practical manufacturing complexity despite the sophisticated design
Solution Approach 2:
The patent creates composite structures by combining binding proteins with fluorescent labels through genetic fusion or chemical conjugation. These composite fluorescent-protein constructs achieve both specific binding capability and detectable signal, enabling accurate detection while the modular nature of the composite allows for standardized production protocols
3Adaptability or versatility
If fluorescent sensors are used to detect and discriminate multiple microbe-associated compounds, then detection capability improves, but the device complexity increases
Solution Approach 1:
The patent segments the detection task by using different fluorescently labeled binding proteins, each specific to a particular class of microbe-associated compounds (e.g., siderophores, vitamins, enzymes). This segmentation allows multiple compounds to be detected simultaneously through a panel of specialized sensors, improving versatility while keeping each individual sensor relatively simple
Solution Approach 2:
The patent employs a universal detection platform based on fluorescence measurement that can detect various types of microbe-associated compounds using different binding proteins. The fluorescent detection system serves multiple functions across different compound classes, allowing the same basic assay technology to be applied universally to detect siderophores, vitamins, enzymes, and other microbial metabolites
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
These sensors enable accurate and sensitive detection of microbes in various samples, providing information on microbial identity and potential pathogenicity, and can be used in conjunction with other assays for further analysis and treatment identification.
Implementation Method 1
a high affinity binding protein engineered with a detectable label that generates a detectable signal. The assay solution is exposed to an energy source to generate the initial detectable signal
Data Source
AI summary
Methods, kits, and devices for detecting microbes in various samples. The methods, kits, and devices utilize a high affinity protein-based sensor, wherein the high affinity protein-based sensor comprises a high affinity binding protein and/or bacteria engineered with such a protein, with a detectable label that generates a detectable signal. The high affinity binding protein is specific for a microbe-associated compound secreted or produced by the target microbe When a biological sample containing or suspected of containing a microbe is contacted with the sensor, the presence of the microbe can be detected due to changes in the detectable signal in the assay over time, which correspond to interaction of the microbe-associated compound with the high affinity binding protein.