Adiponectin Multimer Dissociation for Accurate Immunological Assay

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Solution Overview

Problem

Current methods for measuring total adiponectin levels in biological samples are cumbersome, lack automation, and fail to accurately account for varying multimer proportions, leading to inaccurate results.

Innovation Solution

A method involving pretreatment with a reducing agent, acid, or protease, followed by polyacrylamide gel electrophoresis, which converts adiponectin multimers into a form with uniform reactivity, allowing for accurate immunological assays without boiling the sample.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If boiling treatment with SDS is used to expose antibody-recognizing sites, then measurement precision is improved, but device complexity and ease of operation deteriorate due to requiring special apparatus and manual handling

Engineering Contradiction:

Improvetotal adiponectin measurement accuracyVSAvoidapparatus requirement for boiling treatment

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent changes the physical state parameters of adiponectin multimers by adjusting pH and ionic strength through buffer solutions, causing dissociation of multimers into monomers without thermal denaturation. This allows antibody recognition sites to be exposed while avoiding the need for boiling apparatus and complex temperature control systems.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent replaces the mechanical/thermal system (boiling with SDS) with a chemical system (pH and ionic strength adjustment). The buffer solution chemically induces multimer dissociation through electrostatic interactions, eliminating the need for thermal energy input and special boiling apparatus.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Ease of operation

If competitive reaction method with polyclonal antibody is used, then ease of operation is improved, but measurement precision deteriorates due to varying multimer proportions affecting reactivity

Engineering Contradiction:

Improvehandling convenienceVSAvoidtotal adiponectin measurement accuracy

Core Design Contradiction:

Ease of operationVSMeasurement precision

Solution Approach 1:

The patent performs preliminary dissociation of adiponectin multimers into monomers using buffer treatment before the immunoassay step. This preliminary action ensures that all adiponectin molecules are in a uniform monomeric state with accessible antibody-binding sites, eliminating the variability caused by different multimer proportions and enabling accurate total adiponectin measurement.

Inventive Principle:

Principle #10Preliminary action

3Ease of operation

If monoclonal antibody recognizing non-denatured adiponectin is used, then ease of operation is improved, but measurement precision deteriorates because the form of adiponectin affects reactivity

Engineering Contradiction:

Improveassay simplicityVSAvoidtotal adiponectin measurement accuracy

Core Design Contradiction:

Ease of operationVSMeasurement precision

Solution Approach 1:

The patent changes the structural parameters of adiponectin from multimeric to monomeric form through buffer-induced dissociation. This parameter change creates a uniform molecular state that maintains antibody reactivity while eliminating the interference of varying multimer proportions, thereby improving measurement precision without requiring complex denaturation procedures.

Inventive Principle:

Principle #35Parameter changes

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 convenient, quick, and accurate determination of total adiponectin levels in biological samples by converting multimers into a form that maintains reactivity with antibodies, overcoming the limitations of existing methods.

Implementation Method 1

adding, to an adiponectin-containing sample, at least one of a reducing agent, an acid or a salt thereof, a surfactant, and a protease

Methodology Applied

Scientific EffectDisulfide bond reduction: Reduction

Implementation Method 2

adding, to an adiponectin-containing sample, at least one of a reducing agent, an acid or a salt thereof, a surfactant, and a protease

Methodology Applied

Scientific EffectProtonation:

Implementation Method 3

adding, to an adiponectin-containing sample, at least one of a reducing agent, an acid or a salt thereof, a surfactant, and a protease

Methodology Applied

Scientific EffectSurfactant action: Surfactant

Implementation Method 4

adding, to an adiponectin-containing sample, at least one of a reducing agent, an acid or a salt thereof, a surfactant, and a protease

Methodology Applied

Scientific EffectProteolytic cleavage: Enzyme

Implementation Method 5

followed by polyacrylamide gel electrophoresis

Methodology Applied

Scientific EffectGel electrophoresis: Electrophoresis

Data Source

PatentUS7781171B2Mehod of pretreating sample and immunological assay method using the same

Publication Date: 2010.08.24 SEKISUI MEDICAL CO LTD

AI summary

The invention provides a method of pretreating a sample for conveniently, quickly and accurately measuring the total amount of adiponectin present in a biological sample contaminated with various adiponectin multimers.The method of measuring an sample for immunologically assaying the total amount of adiponectin present in the sample comprises reacting, with an adiponectin-containing sample, at least one of a reducing agent, an acid or a salt thereof, a surfactant, and a protease.