Nucleic Acid Aptamer for Beta-Lactoglobulin Detection
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Solution Overview
Problem
Current methods for detecting β-lactoglobulin in cow milk and dairy products are costly, have limited applicability, and suffer from poor specificity and high interference factors, with existing nucleic acid aptamers having high dissociation constants and unclear binding mechanisms.
Innovation Solution
A nucleic acid aptamer specifically recognizing β-lactoglobulin with a sequence having 60% or higher homology to SEQ ID NO:1, modified with various chemical groups, is used in a detection kit that involves a recognition probe and beacon hairpin molecule for high-sensitivity and low-cost detection, with a dissociation constant below 100 nM and specific binding to β-lactoglobulin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If HPLC is used for detection of β-lactoglobulin, then detection accuracy is improved, but cost and device complexity increase
Solution Approach 1:
The patent replaces the complex mechanical HPLC system with a biosensor system that uses biological recognition elements (aptamers) combined with simple detection methods such as colorimetric or fluorescent assays. This substitution maintains high detection accuracy while eliminating the need for expensive and complex HPLC instrumentation.
Solution Approach 2:
The patent uses aptamers as synthetic copies that mimic the function of natural antibodies in recognizing β-lactoglobulin. These aptamers can be produced through in vitro selection (SELEX) and serve as simplified alternatives to complex protein-based detection systems, achieving comparable specificity without the complexity of antibody production and handling.
2Measurement precision
If qPCR is used for detection of β-lactoglobulin, then detection sensitivity is improved, but operation complexity increases
Solution Approach 1:
The patent replaces the complex qPCR amplification system with a direct binding assay using aptamers. The detection is achieved through simple incubation of the aptamer with the sample followed by straightforward readout methods, eliminating the need for thermal cycling, DNA polymerases, and complex data analysis required by qPCR.
3Ease of manufacture
If ELISA is used for detection of β-lactoglobulin, then detection cost is reduced, but detection sensitivity and stability worsen
Solution Approach 1:
The patent changes the recognition element from protein-based antibodies to nucleic acid-based aptamers. This parameter change allows the system to maintain the cost-effectiveness and simplicity of ELISA while achieving superior sensitivity and stability characteristics of nucleic acid-based systems. The aptamers can be synthesized chemically at low cost and exhibit enhanced stability compared to protein antibodies.
4Ease of manufacture
If existing nucleic acid aptamers are used for β-lactoglobulin detection, then detection cost is reduced, but affinity and specificity worsen
Solution Approach 1:
The patent performs preliminary optimization of aptamer sequences and structures before deployment in detection assays. This includes in vitro selection (SELEX) to identify high-affinity binders, structural characterization to ensure proper folding, and validation against structurally related proteins to confirm specificity. This preliminary optimization ensures that the low-cost aptamer-based system achieves reliability comparable to or exceeding more expensive methods.
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 aptamer provides a low-cost, high-specificity detection tool for β-lactoglobulin, capable of distinguishing structural analogs, with a stable stem-loop structure and simulated binding mode, offering improved affinity and specificity for β-lactoglobulin in cow milk and dairy products.
Implementation Method 1
By means of spatial structure matching, hydrogen bonding, van der Waals force, electrostatic interactions, and other intermolecular interactions, the aptamer binds to a target substance with high-affinity
Implementation Method 2
By means of spatial structure matching, hydrogen bonding, van der Waals force, electrostatic interactions, and other intermolecular interactions, the aptamer binds to a target substance with high-affinity
Implementation Method 3
By means of spatial structure matching, hydrogen bonding, van der Waals force, electrostatic interactions, and other intermolecular interactions, the aptamer binds to a target substance with high-affinity
Implementation Method 4
By means of spatial structure matching, hydrogen bonding, van der Waals force, electrostatic interactions, and other intermolecular interactions, the aptamer binds to a target substance with high-affinity
Data Source
AI summary
The present invention provides a nucleic acid aptamer specifically recognizing β-lactoglobulin and use thereof. The nucleic acid aptamer has a sequence as shown in SEQ ID NO:1, a sequence having 60% or higher homology to the sequence as shown in SEQ ID NO:1 and specifically recognizing β-lactoglobulin, or a sequence derived from the sequence as shown in SEQ ID NO:1 and specifically recognizing β-lactoglobulin. The nucleic acid aptamer specifically binds to the allergen β-lactoglobulin in cow milk and dairy products, thereby providing a new tool for the high-sensitivity and low-cost detection of the allergen β-lactoglobulin.