Double Antigen Bridging Immunoassay for Anti-Drug Antibody Differentiation

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Solution Overview

Problem

Current anti-drug antibody assays face challenges in distinguishing between specific and non-specific antibodies, particularly in identifying monomeric and oligomeric forms, leading to interference from other antibodies in the sample, which affects the accuracy of immunoassays.

Innovation Solution

A method using a double antigen bridging immunoassay with a capture drug antibody conjugated to a solid phase and a tracer drug antibody conjugated to a detectable label, where the sample is spiked with monomeric and oligomeric forms of the drug antibody and human IgG to determine the presence and form of anti-drug antibodies, allowing for classification of responses as specific or non-specific and monomeric or oligomeric.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If standard solid-phase immunoassays are used to detect anti-drug antibodies, then the assay can identify the presence of antibodies, but it cannot distinguish between specific and non-specific antibodies or determine their monomeric/oligomeric forms

Engineering Contradiction:
Improvespecificity of anti-drug antibody detectionVSAvoidinformation about antibody form and specificity
Core Design Contradiction:
Measurement precisionVSLoss of information

Solution Approach 1:

The assay is divided into multiple separate testing conditions: samples are tested both with and without spiking with monomeric drug antibody, and both with and without spiking with oligomeric drug antibody. This segmentation allows independent evaluation of each condition to determine antibody specificity and form based on signal changes across the different test configurations.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

Monomeric and oligomeric forms of the drug antibody are prepared in advance as spiking standards. These pre-prepared standards are then used to spike samples before assay testing, allowing the sample to be tested under controlled conditions that reveal the form and specificity characteristics of any anti-drug antibodies present.

Inventive Principle:
Principle #10Preliminary action

2Measurement precision

If the sample is spiked with monomeric and oligomeric forms of drug antibody to determine antibody form, then the measurement precision improves, but the device complexity increases

Engineering Contradiction:
Improvedetermination of antibody formVSAvoidcomplexity of immunoassay protocol
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The same immunoassay system and reagents are used for multiple purposes: detecting the presence of anti-drug antibodies, determining whether the antibodies are specific or non-specific, and identifying whether they are monomeric or oligomeric forms. The spiking controls serve multiple functions simultaneously, allowing one assay system to perform what would traditionally require multiple separate assays.

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Ease of operation

If conventional immunoassays are used without spiking controls, then the ease of operation is maintained, but false positives occur due to interference from other antibodies in the sample

Engineering Contradiction:
Improvesimplicity of immunoassay procedureVSAvoidaccuracy of anti-drug antibody detection
Core Design Contradiction:
Ease of operationVSReliability

Solution Approach 1:

Monomeric and oligomeric forms of the drug antibody are spiked into the sample before the immunoassay to preemptively identify and account for interfering antibodies. By introducing these controls in advance, the assay can distinguish whether a positive signal is due to the target anti-drug antibody or to interfering antibodies in the sample, thereby preventing false positives while maintaining the simplicity of the overall procedure.

Inventive Principle:
Principle #9Preliminary anti-action

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 approach enables accurate differentiation of anti-drug antibodies from non-specific antibodies, reducing false positives and improving the specificity and sensitivity of immunoassays, particularly for therapeutic antibodies like those targeting IL-6 receptor, IGF-1 receptor, or IL-13 receptor 1 alpha.

Implementation Method 1

contacting said capture drug antibody separately with b-i) said sample

Methodology Applied
Scientific EffectAntigen-antibody interaction:

Implementation Method 2

said tracer drug antibody, which is said drug antibody conjugated to a detectable label

Methodology Applied
Scientific EffectEnzyme-label conjugation:

Implementation Method 3

The activity of the antibody-conjugated enzyme is proportional to the antigen concentration in the incubation medium

Methodology Applied
Scientific EffectSignal detection:

Implementation Method 4

a sandwich is formed: solid phase-capture antibody-antigen-tracer antibody

Methodology Applied
Scientific EffectSandwich immunoassay:

Implementation Method 5

The standard sandwich method is also called double antigen bridging immunoassay because capture and tracer antibodies bind to different epitopes of the same antigen

Methodology Applied
Scientific EffectDouble antigen bridging:

Data Source

PatentEP2573568B1Distinguishing assay
Publication Date: 2015.01.14 F HOFFMANN LA ROCHE & CO AG
  • EP2573568B1 patent drawingFigure 1

AI summary

The current invention comprises a method for determining of an antibody against a drug antibody in a sample using an immunoassay comprising a capture drug antibody and a tracer drug antibody, wherein the method comprises providing i) a capture drug antibody, which is the drug antibody conjugated to a solid phase, ii) a tracer drug antibody, which is the drug antibody conjugated to a detectable label, contacting the capture drug antibody separately with i) the sample, ii) the sample, to which the drug antibody in monomeric form has been added, iii) the sample, to which the drug antibody in oligomeric form has been added, and determining an antibody against the drug antibody in the sample by a positive immunoassay in i) and a negative immunoassay in ii) and iii).