S100A12 Mutant Standards for Dimer-Specific Immunoassays

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Solution Overview

Problem

Current methods for detecting S100A12 proteins in biological samples cannot differentiate between dimers, tetramers, and hexamers, making it difficult to accurately diagnose and monitor inflammatory diseases, as only the dimeric form is physiologically active and indicative of inflammatory conditions.

Innovation Solution

Development of S100A12 mutants with mutations in the high- or low-affinity calcium binding regions or zinc binding region that can only form dimers, allowing for the reliable detection and quantification of S100A12 dimers in samples, which can be used as standards in immunoassays to differentiate between dimeric, tetrameric, and hexameric forms.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If conventional S100A12-ELISAs are used for quantification, then the measurement can be performed using diverse antibodies, but it cannot be differentiated whether the measured values represent the dimer, tetramer, or hexamer form

Engineering Contradiction:

Improveuse of diverse antibodiesVSAvoiddifferentiation between oligomeric forms

Core Design Contradiction:

Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The patent applies local quality by creating mutants with specific modifications in particular regions (high-affinity calcium binding hand, low-affinity calcium binding hand, or zinc binding region) that confer the ability to form only dimers. This localized modification achieves the goal of oligomeric form differentiation while maintaining antibody binding capability.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent changes the molecular parameters of S100A12 by introducing mutations in calcium binding or zinc binding regions, which alters the protein's oligomerization behavior. This parameter change transforms the protein from forming multiple oligomeric states to forming exclusively dimers, enabling precise measurement differentiation.

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If S100A12 mutants with mutations in calcium binding regions or zinc binding region are used, then dimer formation is restricted and reliable detection is enabled, but the complexity of the method increases

Engineering Contradiction:

Improvereliable detection of S100A12 dimersVSAvoidcomplexity of mutant generation and detection method

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent applies preliminary action by pre-generating standardized S100A12 mutants with defined mutations in calcium binding or zinc binding regions. These pre-prepared mutants serve as reliable standards that can be directly used in detection methods, eliminating the need for complex real-time oligomeric form analysis during measurement.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent uses S100A12 mutants as intermediary standards that mediate between the complex biological samples containing mixed oligomeric forms and the detection system. These mutants provide a simplified, well-defined reference that bridges the gap between sample complexity and measurement simplicity.

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

Enables precise detection and quantification of biologically active S100A12 dimers, providing a reliable diagnostic tool for acute and chronic inflammatory diseases, allowing for early detection of subclinical residual activities and monitoring of disease progression.

Implementation Method 1

S100A12 mutants having at least one mutation in the high- or low-affinity calcium binding hand or the zinc binding region of S100A12 are no longer able to tetramerize or hexamerize and can thus be applied as a reliable standard for the detection of S100A12 dimers in a sample

Methodology Applied

Scientific EffectCalcium binding:

Implementation Method 2

S100A12 proteins are usually only found in oligomeric states, such as dimers, tetramers, or hexamers. Further, these different oligomeric states appear to be related to different functional states. It is assumed that the dimeric form of S100A12 is particularly relevant for the diagnosis or assessment of the disease state

Methodology Applied

Scientific EffectProtein dimerization:

Data Source

PatentUS11267854B2Complex-specific standardization of immunological methods for the quantification of S100A12

Publication Date: 2022.03.08 WESTFAELISCHE WILHELMS-UNIVERSITAET MUENSTER

AI summary

The present invention relates to mutants of S100A12 having at least one mutation in the high-affinity calcium binding hand or the low-affinity calcium binding hand or the zinc binding region. The present invention also relates to methods of detecting S100A12 dimers in a sample as well as methods of diagnosis using the S100A12 mutant of the invention, as well as to diagnostic compositions and kits comprising such an S100A12 mutant. The present invention further relates to a method of generating an antibody that specifically binds to an S100A12 dimer using the S100A12 mutant of the invention, as well as to an antibody that specifically binds to an S100A12 dimer.