Aqueous Immunoassay Buffer Reducing Unspecific Binding

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Solution Overview

Problem

Current immunoassay buffers fail to effectively reduce unspecific binding, low-affinity binding, cross-reactivities, and matrix effects, leading to false signals and reduced sensitivity, especially in multi-analyte applications and plasma samples.

Innovation Solution

An aqueous solution comprising a buffer with a compound defined by the formula R1—[[CR2R3]p—O]q—R4, polyol, saccharide, and a non-ionic detergent, which controls pH and reduces unspecific interactions, cross-reactivities, and matrix effects, while enhancing the specific binding activity of antibodies.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional immunoassay buffers (PBS with BSA and Tween 20) are used, then the basic binding function is maintained, but unspecific binding, cross-reactivities and matrix effects cannot be avoided

Engineering Contradiction:

Improvespecific binding activityVSAvoidunspecific binding and cross-reactivities

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention changes the chemical composition parameters of the buffer by incorporating polyols (glycerol, sorbitol, mannitol) and saccharides (sucrose, trehalose) in specific concentrations (0.1-10% w/v). This parameter modification fundamentally alters the buffer's ability to suppress unsspecific binding while maintaining pH control and solubility, directly resolving the contradiction between maintaining specific binding and eliminating harmful unspecific interactions.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention creates a composite buffer system that combines multiple functional components: pH control agents (phosphates, citrates, acetates), blocking agents (BSA, casein, gelatin), non-ionic detergents (Tween 20, Triton X-100), and newly added polyols/saccharides. This composite approach synergistically addresses both specific binding maintenance and unsific binding suppression, overcoming the limitations of simple PBS formulations.

Inventive Principle:

Principle #40Composite materials

2Object-affected harmful factors

If detergents are added to buffers for washing steps, then unspecific bindings are removed, but sensitivity and detection precision are reduced due to persistent matrix effects

Engineering Contradiction:

Improveunspecific binding removalVSAvoiddetection sensitivity

Core Design Contradiction:

Object-affected harmful factorsVSMeasurement precision

Solution Approach 1:

The invention applies blocking agents and polyols/saccharides to the buffer before the binding reaction occurs. This preliminary action pre-coats surfaces and occupies potential unsific binding sites, preventing harmful interactions from occurring in the first place. By addressing unsific binding prevention proactively rather than reactively through washing, the invention maintains detection sensitivity while eliminating matrix effects.

Inventive Principle:

Principle #10Preliminary action

3Adaptability or versatility

If multiple antibodies are used for multi-analyte detection, then assay versatility increases, but cross-reactivities between antibodies become a problem

Engineering Contradiction:

Improvemulti-analyte detection capabilityVSAvoidcross-reactivity

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The invention modifies the buffer composition parameters by adding polyols and saccharides that specifically suppress antibody cross-reactivity. These additives change the chemical environment in a way that reduces non-specific antibody-antigen interactions while preserving specific high-affinity binding. This allows multiple antibodies to be used in the same assay system without cross-reactivity interference, enabling reliable multi-analyte detection.

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

The solution significantly reduces unspecific binding, cross-reactivities, and matrix effects, increasing the sensitivity of immunoassays by enhancing the specific binding activity of antibodies and preventing negative effects from heterophilic antibodies, rheuma factors, hemoglobin, bilirubin, and triglycerides.

Implementation Method 1

a buffer to control pH

Methodology Applied

Scientific EffectpH control:

Implementation Method 2

a compound A selected from the group consisting of: a compound defined by the general formula I R1—[[CR2R3]p—O]q—R4... with the proviso that the compound at least carries two hydroxy groups; polyol; saccharide

Methodology Applied

Scientific EffectMolecular recognition and steric hindrance:

Implementation Method 3

a non-ionic detergent

Methodology Applied

Scientific EffectSurfactant action: Surfactant

Data Source

PatentUS8877514B2Aqueous solution for use as medium for the specific binding reaction of a binding pair

Publication Date: 2014.11.04 CANDOR BIOSCI

AI summary

The present invention refers to an aqueous solution for use as medium for the specific binding reaction of a binding pair, wherein a first binding member recognizes its complementary second binding member. The solution contains a) a buffer to control pH; b) a compound A selected from a compound defined by the general formula I R1—[[CR2R3]P—O]q—R4, wherein R1 is hydrogen or hydroxy group, R2 for each unit independently is hydrogen or hydroxy group, R3 is hydrogen, methyl group, or ethyl group, R4 is hydrogen or alkyl group, p is an integer of from 2 to 10 and q is an integer of from 1 to 100, with the proviso that the compound at least carries two hydroxy groups; a polyol; or saccharide; and c) a non-ionic detergent.