Leucomalachite Green Immunoassay Direct Detection

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Solution Overview

Problem

Current methods for detecting and determining leucomalachite green and malachite green in samples are limited, particularly due to the difficulty in analyzing leucomalachite green, which lacks chromophores in the visible region, requiring additional steps like oxidation to malachite green for detection, and existing immunoassays are time-consuming and prone to errors.

Innovation Solution

Development of immunogens and antibodies specific to leucomalachite green, coupled with crosslinkers to antigenicity-conferring carrier materials or detectable labeling agents, allowing for direct detection and quantification of both leucomalachite green and malachite green without the need for pre-treatment, enhancing sensitivity and reducing experimental errors.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Difficulty of detecting and measuring

If post-column oxidation step is used to detect leucomalachite green, then detection capability is improved, but analysis time and process complexity increase

Engineering Contradiction:
Improvedetection capabilityVSAvoidanalysis time
Core Design Contradiction:
Difficulty of detecting and measuringVSLoss of time

Solution Approach 1:

The patent applies preliminary action by developing antibodies that can directly recognize and bind to leucomalachite green in its reduced form, eliminating the need for post-column oxidation steps. The immunogen is designed with a crosslinker attached to the phenyl ring position that mimics the natural epitope structure, enabling direct detection without chemical conversion, thus saving analysis time while maintaining detection capability.

Inventive Principle:
Principle #10Preliminary action

2Measurement precision

If lead dioxide or electrochemical oxidation is used for pre-column oxidation, then detection sensitivity is improved, but method complexity and potential contamination increase

Engineering Contradiction:
Improvedetection sensitivityVSAvoidmethod complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts the oxidation step entirely from the analytical method by developing antibodies specific to leucomalachite green. Instead of requiring chemical oxidation reagents like lead dioxide or electrochemical cells, the method directly detects the reduced form using antigen-antibody binding, thereby simplifying the methodology while maintaining or improving detection sensitivity through specific immunological recognition.

Inventive Principle:
Principle #2Taking out (Extraction)

3Measurement precision

If radioimmunoassay is used for detection, then sensitivity is improved, but safety risks and regulatory requirements increase

Engineering Contradiction:
ImprovesensitivityVSAvoidsafety risks
Core Design Contradiction:
Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The patent replaces long-lived radioactive isotopes with non-radioactive detection systems based on enzyme-linked or fluorescent labels. These alternative labels provide sufficient sensitivity for trace detection without the safety hazards, regulatory burdens, and long-term waste management issues associated with radionuclides, while maintaining the high sensitivity characteristic of immunoassays.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

4Measurement precision

If multiple analysis steps are used for malachite green and leucomalachite green, then detection accuracy is improved, but productivity decreases

Engineering Contradiction:
Improvedetection accuracyVSAvoidanalysis throughput
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The patent merges the detection of malachite green and leucomalachite green into a single immunoassay procedure. By developing antibodies that recognize the phenyl ring structure common to both compounds and using a unified sample processing protocol, the method achieves accurate simultaneous detection of both analytes without requiring separate oxidation and analysis steps for each compound, thereby significantly improving productivity while maintaining detection accuracy.

Inventive Principle:
Principle #5Merging (Combining)

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 solution enables efficient, sensitive, and specific detection and quantification of leucomalachite green and malachite green, improving upon existing methods by reducing the complexity and increasing the accuracy of the analysis, with high cross-reactivity and sensitivity, allowing for detection at concentrations below established limits.

Implementation Method 1

an immunogen comprising leucomalachite green, coupled by way of a crosslinker, to an antigenicity-conferring carrier material

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Implementation Method 2

antibodies raised against such immunogens... Specific binding reactions, such as antibody-antigen interactions

Methodology Applied
Scientific EffectAntigen-antibody binding:

Implementation Method 3

conjugates comprising leucomalachite green, covalently bonded by way of a crosslinker, to a detectable labelling agent

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Data Source

PatentEP1767947B1An immunoassay method and kit to leucomalachite green and malachite green
Publication Date: 2008.10.29 RANDOX LAB LTD
  • EP1767947B1 patent drawingFigure 1
  • EP1767947B1 patent drawingFigure 2
  • EP1767947B1 patent drawingFigure 3

AI summary

The invention provides haptens, immunogens comprising such haptens coupled to an antigenicity-conferring carrier material, conjugates comprising such haptens bonded to a labelling agent as well as, antibodies raised against such immunogens and capable of binding with leucomalachite green and malachite green.