Fluorescence Assay for Intestinal Permeability Using Organoborane
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Solution Overview
Problem
Current methods for assessing small intestinal permeability are time-consuming and costly, as they rely on sugar markers like lactulose and mannitol that lack intrinsic absorbance or fluorescence, making rapid and direct quantification challenging.
Innovation Solution
A fluorescence assay using an organoborane compound coupled with a fluorophore to measure sugars in biological samples, allowing for the quantification of sugars in biological fluids within 0 to 24 hours post-ingestion, and the development of kits and assays for this method.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional HPLC or enzyme assays are used to measure sugar markers, then measurement precision is achieved, but measurement time and cost increase significantly
Solution Approach 1:
The patent employs fluorogenic substrates as intermediaries that convert non-fluorescent sugar markers into fluorescent compounds through enzymatic reactions. This intermediary conversion enables rapid fluorescence-based detection, replacing time-consuming HPLC or enzyme assays while maintaining quantification accuracy through standardized calibration curves.
Solution Approach 2:
The invention utilizes fluorescence emission as a detectable signal change, where sugar markers are converted into fluorescent compounds that emit light at specific wavelengths. This color/fluorescence change enables rapid, direct quantification through fluorescence readers, dramatically reducing measurement time compared to conventional methods that require HPLC separation or complex enzyme assays.
2Reliability
If conventional methods are used for intestinal permeability assessment, then reliable results are obtained, but throughput is limited and cost per sample is high
Solution Approach 1:
Fluorogenic substrates serve as reliable intermediaries that provide consistent, reproducible fluorescence signals for permeability assessment. The standardized enzymatic conversion and fluorescence detection system ensures reliable results while enabling high-throughput processing of multiple samples simultaneously using plate readers, thereby increasing productivity without compromising measurement reliability.
Solution Approach 2:
The patent replaces complex mechanical separation systems (HPLC) and multi-step enzyme assays with a simplified fluorescence-based detection system. This substitution maintains measurement reliability through controlled enzymatic reactions and standardized fluorescence quantification, while dramatically improving throughput by enabling rapid, automated reading of multiple samples in parallel.
3Measurement precision
If sugar markers without intrinsic fluorescence are measured, then accurate quantification is possible, but detection complexity and cost increase
Solution Approach 1:
The patent introduces fluorogenic substrates as intermediaries that convert non-fluorescent sugar markers into fluorescent compounds. This intermediary step adds minimal complexity while enabling simple fluorescence-based detection, replacing complex HPLC or enzyme assays. The standardized substrate-converter-enzyme system maintains quantification accuracy through calibration curves while significantly simplifying the detection infrastructure.
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
This approach simplifies the measurement of intestinal permeability, increases throughput, and reduces costs by enabling rapid and direct quantification of sugars, providing accurate results comparable to conventional enzyme assays.
Implementation Method 1
measuring the amount of the one or more sugars in the biological sample with an organoborane compound coupled to a fluorophore
Data Source
AI summary
Provided is a method for quantifying sugar appearing in a biological fluid of a subject, the method include (a) obtaining a biological sample from a subject, who has ingested one or more sugars, over a period of 0 to 24 hours after the ingesting; and (b) measuring the amount of the one or more sugars in the biological sample with an organoborane compound coupled to a fluorophore. Also provided are assays and kits for performing the above methods.