Separation-Free Assay Mobile Binding Partner

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Solution Overview

Problem

Current high-throughput screening technologies in drug discovery face challenges with cumbersome and expensive methods that rely on specific binding substances, particularly in sandwich type assays and whole cell assaying, due to distance dependence and labeling requirements, limiting their effectiveness and applicability.

Innovation Solution

A separation-free assay method using a binding partner label and a nonspecifically binding label, where the signal from either label is measured in a binding event, with at least one binding partner being mobile and the other labeled, allowing for direct luminescence and quenching or enhancement of the signal without the need for specific capture molecules or immobile surfaces.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If sandwich type assay systems are used, then specific binding and detection can be achieved, but the distance between donor and acceptor elements becomes too long to generate adequate signal

Engineering Contradiction:

Improvesignal generationVSAvoiddistance between donor and acceptor

Core Design Contradiction:

Measurement precisionVSLength of stationary object

Solution Approach 1:

The invention extracts the detection element from the sandwich structure and places it directly on the mobile binding partner. This eliminates the need for long-distance signal transmission through the sandwich configuration, allowing the detection element to be in direct proximity to the binding site and thus generate adequate signal without distance limitations.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

Instead of having the detection element fixed on the stationary phase and requiring the mobile binding partner to approach it (sandwich configuration), the invention inverts the approach by placing the detection element on the mobile binding partner itself. This reversal eliminates the distance problem inherent in sandwich assays where the detection element must be close to the binding site.

Inventive Principle:

Principle #13The other way round (Inversion)

2Measurement precision

If multiple labeling agents are used for different binding partners, then specific detection can be achieved, but the process becomes extremely cumbersome and expensive

Engineering Contradiction:

Improvespecific detectionVSAvoidlabeling process

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The invention employs a universal labeling strategy where a single type of labeling agent is used for all binding partners. The detection element on the mobile binding partner serves multiple functions: it detects the binding event and provides the signal for measurement. This universal approach simplifies the labeling process compared to using different labeling agents for different binding partners, reducing both complexity and cost while maintaining specific detection capability.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Measurement precision

If specific binding substances are used, then high specificity and affinity can be achieved, but the method becomes less adaptable to different assay formats

Engineering Contradiction:

Improvespecificity and affinityVSAvoidassay format adaptability

Core Design Contradiction:

Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The invention makes the binding partner mobile rather than fixed on a stationary phase. This dynamic approach allows the same basic assay format to be adapted to different applications (e.g., different binding partners, different detection methods) by simply changing the mobile binding partner while keeping the detection element strategy the same. The mobile binding partner can be any molecule that can bind to the analyte, providing versatility across different assay types.

Inventive Principle:

Principle #15Dynamics

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

This method simplifies the detection process, reduces background signal, and enhances sensitivity, enabling efficient and cost-effective analysis of sample substances, including whole cells and nucleic acids, by allowing signal measurement in solution without the need for specific biomolecule coupling to an immobile surface.

Implementation Method 1

a directly luminescent label adsorbed and/or covalently coupled to at least one of said binding partners

Methodology Applied

Scientific EffectLuminescence: Luminescence

Implementation Method 2

said nonspecifically binding label is able to affect the signal of said binding partner label

Methodology Applied

Scientific EffectSignal quenching:

Implementation Method 3

said binding partner label is able to affect the signal of said nonspecifically binding label

Methodology Applied

Scientific EffectSignal enhancement:

Data Source

PatentUS8956877B2Separation-free assay method

Publication Date: 2015.02.17 KEMIRA OY

AI summary

A separation-free assay including a binding partner, a binding partner labeled with a directly luminescent label, and a nonspecifically binding label able to affect the signal of the unbound labeled binding partner, in which a signal of the binding partner label or the signal of the nonspecific binding label is measured in a binding event, and where at least one of the binding partners is a mobile binding partner.