Depleting Multispecific Antibody Complexes for Free Antigen Detection
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Solution Overview
Problem
Current methods for detecting free, non-complexed binding partners in samples are inefficient due to the inability to effectively deplete the bound multispecific binder complexes, which interferes with the accurate determination of free binding partners.
Innovation Solution
The method involves depleting the multispecific binder from the sample by incubating it with a second binding partner or a monospecific binder that specifically binds to a different specificity of the multispecific binder, allowing for the detection of free binding partners by forming complexes with anti-idiotypic antibodies and solid-phase immobilization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If standard solid-phase immunoassays are used to detect binding partners, then the assay can detect total binding partner (free and bound), but the presence of bound complexes interferes with accurate determination of free binding partner
Solution Approach 1:
The patent extracts and removes the bound multispecific binder complexes from the sample before detecting the free binding partner. This is achieved by incubating the sample with a second binding partner or monospecific binder that selectively binds to the multispecific binder, thereby separating the bound complexes from the free binding partner and enabling accurate detection of the free form.
Solution Approach 2:
The patent performs a preliminary depletion step before the actual detection. By pre-incubating the sample with selective binders to remove bound complexes, the detection assay can then accurately measure only the free binding partner without interference from bound forms.
2Reliability
If multispecific binder complexes are not depleted, then the detection assay remains simple, but the accuracy of free binding partner detection is compromised
Solution Approach 1:
The patent introduces intermediary substances (second binding partner or monospecific binder) that mediate the depletion process. These intermediaries selectively bind to the multispecific binder, facilitating its removal from the sample without directly interfering with the detection of the free binding partner.
3Measurement precision
If depletion of bound complexes is performed, then accurate detection of free binding partner is achieved, but additional reagents and steps are required
Solution Approach 1:
The patent uses selective binders in controlled amounts to achieve sufficient depletion of bound complexes. The second binding partner or monospecific binder is used in quantities that are sufficient to remove bound multispecific binders from the sample, ensuring accurate detection of free binding partners while minimizing reagent consumption.
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 precise determination of free binding partners by removing bound complexes, thereby enhancing the accuracy and reliability of detecting free antigens or antibodies in samples.
Implementation Method 1
a multispecific binder which has a first binding specificity that specifically binds to a first binding partner
Implementation Method 2
incubating the sample with a second binding partner that can be specifically bound by a second binding specificity of the multisspecific binder
Data Source
AI summary
Herein is reported a method for the detection of free antigen of a multispecific antibody in a sample, whereby the antigen to be detected can be specifically bound by a first binding site of the multispecific antibody, comprising the step of incubating a sample comprising free antigen and multispecific antibody with an anti-idiotypic antibody, which specifically binds to a second binding specificity of the bispecific antibody, which is different from the first binding specificity, whereby the anti-idiotypic antibody is bound to a solid phase.