Enzyme Inactivation for Antimicrobial Sample Analysis

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Solution Overview

Problem

Existing methods for analyzing samples for vaccine efficacy are hindered by the presence of antimicrobial substances, which can interfere with microbial viability assays and lead to incorrect results or the discard of usable samples.

Innovation Solution

A method involving the use of an enzyme, specifically a New Delhi metallo-beta-lactamase 1 (NDM-1) polypeptide, to inactivate antimicrobial substances in samples, allowing for the preparation of samples for analysis without interference from these substances.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If samples from vaccinated subjects are analyzed for vaccine efficacy, then vaccine protective effect can be determined, but antimicrobial substances in the sample may interfere with the analysis and lead to incorrect results or sample discard

Engineering Contradiction:
Improveanalysis result accuracyVSAvoidantimicrobial substance interference
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent applies preliminary action by treating the sample with an enzyme (such as beta-lactamase) before the actual analysis to inactivate antimicrobial substances. This pre-treatment step ensures that the harmful substances are neutralized prior to the assay, preventing interference with the analysis of vaccine efficacy while maintaining the integrity of the original sample for its intended purpose.

Inventive Principle:
Principle #10Preliminary action

2Measurement precision

If samples containing antimicrobial substances are discarded to ensure analysis accuracy, then correct analysis results can be obtained, but usable samples are lost and clinical study costs increase

Engineering Contradiction:
Improveanalysis result accuracyVSAvoidsample utilization rate
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The patent converts the harmful presence of antimicrobial substances in samples into a beneficial situation by using these substances as indicators to trigger the enzyme treatment. Instead of discarding samples containing antibiotics, the method recognizes their presence and applies targeted enzymatic inactivation, thereby turning what would be a disqualifying factor into a cue for sample preparation, increasing overall sample utilization and reducing study costs.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

3Adaptability or versatility

If enzyme treatment is applied to inactivate antimicrobial substances, then sample usability is improved, but additional treatment steps and time are required

Engineering Contradiction:
Improvesample usabilityVSAvoidsample preparation process complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent introduces an enzyme (such as beta-lactamase or other antimicrobial inactivating enzymes) as an intermediary substance that mediates between the harmful antimicrobial agents and the analysis process. The enzyme specifically targets and inactivates the antimicrobial substances through enzymatic reaction, providing a selective and efficient means of neutralization that adds minimal complexity compared to alternative methods such as extensive filtration or chemical neutralization protocols.

Inventive Principle:
Principle #24Intermediary (Mediator)

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 enables the use of samples that would otherwise be unusable due to antimicrobial contamination, ensuring accurate analysis results and reducing the need for additional samples or subject removal from clinical studies.

Implementation Method 1

The enzyme used in the method is preferably a beta-lactamase, more preferably a carbapenemase, even more preferably a class B carbapenemase. Beta-lactamases are a group of enzymes that provide bacterial resistance to beta-lactam antibiotics by breaking the antibiotic's structure by hydrolysis.

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

treating said sample with an enzyme to inactivate said antimicrobial substance

Methodology Applied
Scientific EffectEnzymatic inactivation: Enzyme

Data Source

PatentEP4553166A1Method for preparing a sample for analysis
Publication Date: 2025.05.14 MINERVAX
  • EP4553166A1 patent drawingFigure 1~2
  • EP4553166A1 patent drawingFigure 3A
  • EP4553166A1 patent drawingFigure 3B

AI summary

The technology proposed herein relates to a method for preparing a sample for analysis, said method comprising: a) providing a sample potentially containing an antimicrobial substance, such as an antibiotic; and b) treating said sample with an enzyme to inactivate said antimicrobial substance. A method comprising a microbial viability assay of a sample, a New Delhi metallo-beta-lactamase 1 (NDM-1) construct for recombinant expression thereof, a method for recombinantly producing a New Delhi metallo-beta-lactamase 1 (NDM-1) protein, and a use of an antimicrobial inactivating enzyme for preparing a sample for an analysis are also disclosed.