Hapten-Immobilized Probe Regeneration via Acidic Elution
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Solution Overview
Problem
Current immunoassay methods are costly and inefficient, particularly when it comes to serial measurements, as they require significant amounts of reagents and can limit clinical testing due to cost constraints.
Innovation Solution
A method that re-uses a hapten-immobilized test probe by dipping it in an acidic solution with a pH of 1-4 after each cycle, allowing for the detection of an analyte in multiple liquid samples while minimizing reagent consumption.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a probe is reused multiple times in immunoassay cycles, then cost is reduced and productivity is improved, but the binding activity and assay performance deteriorate due to repeated exposure to denaturation conditions
Solution Approach 1:
The patent applies parameter changes by carefully controlling the pH conditions during probe regeneration. The probe is exposed to acidic conditions (pH 1-4) for a limited time to elute the immune complex, then rapidly neutralized to restore binding activity. This dynamic adjustment of pH parameters allows the probe to withstand multiple cycles without permanent denaturation, resolving the contradiction between reusability and performance maintenance.
2Ease of repair
If denaturation agents are used to dissociate immune complexes for probe recycling, then the probe can be regenerated, but the antibody charge, hydration, hydrogen bonding and tertiary structure are altered, reducing binding activity
Solution Approach 1:
The patent converts the harmful effect of acid denaturation into a beneficial regeneration process. By using controlled acidic conditions (pH 1-4) to elute the immune complex and then rapidly neutralizing the probe, the temporary structural disruption is transformed into a useful regeneration step. The antibody structure is temporarily altered to release the complex, then restored for the next cycle, turning potential damage into a functional advantage.
3Reliability
If fresh reagents are used for each immunoassay cycle, then assay performance is maintained, but manufacturing cost increases
Solution Approach 1:
The patent implements discarding and recovering by eluting the immune complex from the probe using acidic conditions, then regenerating the probe for reuse. Instead of discarding the probe after a single use, the immune complex is selectively removed and the probe is restored to its binding-active state. This recovery process allows the same probe to be used multiple times, significantly reducing reagent consumption while maintaining assay performance through the dual antibody conjugate system.
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 enables the detection of analytes in multiple samples using the same probe and reagents, significantly reducing costs and maintaining clinical assay performance across multiple cycles.
Implementation Method 1
dipping the probe tip in an acidic solution having pH about 1.0-4.0 to elute the immunocomplex from the probe tip
Implementation Method 2
the signal antibody is conjugated with fluorescent labels; determining the analyte concentration in the sample by measuring the fluorescent signal of the immunocomplex at the probe tip
Data Source
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AI summary
The present invention is directed immunoassay methods, which re-use a hapten-immobilized test probe and reagents for quantitating an analyte in different samples, anywhere from about 3 to 20 times, while maintaining acceptable clinical assay performance. The methods use a dual antibody conjugate solution comprising an anti-hapten antibody and a capture antibody against the analyte in each cycle. After the completion of each cycle of reaction, the test probe is dipped in an acidic solution having pH about 1-4, to elute the immunocomplex formed on the probe and to regenerate the hapten-immobilized probe. The robustness of the hapten-coated solid phase allows utilization of multiple denaturation reagents for efficient elution of the immune complexes after each cycle without compromising the binding activity of the hapten on the solid phase.