Cell-free biosensors to detect creatinine, creatine, and sarcosine
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing diagnostics for quantifying biomarkers in complex biological samples, such as blood or urine, suffer from high inter-sample variability due to matrix effects, making them inaccurate and costly, and are limited to presence/absence detection without quantitative measurement capabilities, especially in minimal-equipment settings.
Innovation Solution
A method using a cell-free expression system with a reporter system, including a regulator and reporter gene, generates a colorimetric gene product that corresponds to specific analyte levels, allowing for sample-specific calibration to reduce variability and enable quantitative measurement of creatinine, creatine, or sarcosine in biological samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If standard quantification approaches with calibration curves are used, then measurement accuracy is improved, but inter-sample variability due to matrix effects increases
Solution Approach 1:
The system uses the sample's own matrix components as the calibration standard. By performing the calibration reaction directly in the sample matrix rather than using external standards, the assay automatically compensates for matrix effects. The sample serves both as the analyte source and the calibration reference, eliminating the discrepancy between standard and sample conditions.
Solution Approach 2:
A regulatory molecule serves as an intermediary between the analyte and the reporter signal. The regulatory molecule's concentration is titrated to achieve a desired dynamic range, and it mediates the interaction between the analyte and reporter system. This intermediary allows the system to normalize responses across different samples by adjusting the regulator concentration in each sample-specific calibration.
2Measurement precision
If sophisticated analytical equipment is used, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The patent replaces complex mechanical and electronic analytical systems with a biochemical system. Instead of using sophisticated instrumentation for quantification, the system uses a cell-free expression system with regulatory molecules and reporter genes that naturally provide quantitative information through measurable biochemical responses. This substitution eliminates the need for expensive equipment while maintaining measurement precision.
Solution Approach 2:
The system uses colorimetric reporter genes that produce visible color changes corresponding to analyte concentrations. This allows quantitative measurement without sophisticated detection equipment, as the color intensity or hue can be measured with simple spectrophotometry or even visual assessment. The color change provides a direct, equipment-free readout of the quantitative analysis.
3Ease of operation
If cell-free expression systems are used for diagnostics, then ease of operation and cost are improved, but quantitative measurement capability is limited
Solution Approach 1:
The system dynamically adjusts the concentration of regulatory molecules to optimize the dynamic range for each specific analyte and sample type. Rather than using fixed parameters, the regulatory molecule concentration is titrated and optimized for each application, allowing the simple cell-free system to achieve quantitative measurement capabilities. This dynamic parameter adjustment enables the system to adapt to different analytes while maintaining ease of operation.
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 provides a cost-effective, equipment-free diagnostic tool that reduces inter-sample variability, enabling accurate quantification of analytes like creatinine and sarcosine, suitable for point-of-care use and low-resource settings, by generating a visible color gradient that can be easily interpreted visually.
Implementation Method 1
a colorimetric gene product is generated having a color corresponding to a color of a first reference point
Data Source
AI summary
An exemplary embodiment of the present disclosure provides a biosensor that can facilitate detection of the molecules creatinine, creatine and/or sarcosine in a minimal-equipment, portable, and low-cost fashion. Some aspects of the disclosure relate to methods and systems to enable quantitative measurement of certain biomarkers at the point of care without expensive instrumentation.