VeraTag Assay for Cell Surface Moiety Clustering Detection
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Solution Overview
Problem
Current methods for detecting and quantifying the expression of multiple cell surface moieties, particularly CD137 and PD-L1, lack robustness in demonstrating their clustering in the presence of multispecific agents, which is crucial for predicting patient response to therapies targeting these antigens.
Innovation Solution
A method involving a VeraTag assay that uses binding molecules with molecular tags and cleavage-inducing moieties to detect and quantify the clustering of CD137 and PD-L1 by inducing cleavage and measuring the presence or absence of released tags, allowing for the assessment of therapeutic agent binding and mode of action.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional IHC or FISH assays are used to detect cell surface moiety expression, then expression levels can be measured, but the ability to demonstrate clustering of multiple antigens in the presence of multispecific agents is lacking
Solution Approach 1:
The patent uses a multispecific agent as an intermediary that simultaneously binds to multiple cell surface moieties (e.g., CD137 and PD-L1), bringing them into proximity to enable clustering detection. This intermediary approach allows the assay to detect clustering that would otherwise be undetectable with traditional single-antigen methods
Solution Approach 2:
The assay employs a nested structure where binding molecules with molecular tags are used to detect the presence and clustering of multiple antigens. The molecular tags are nested within the binding molecules, and the entire system is nested within the cellular context, allowing hierarchical detection of clustering events
2Measurement precision
If FISH assays are used to quantify gene copy number, then quantitative data is obtained, but the assays are costly, labor-intensive, and require advanced training
Solution Approach 1:
The patent replaces the complex fluorescent microscopy system used in FISH assays with a simpler detection system using molecular tags and binding molecules. This substitution maintains quantitative measurement capability while eliminating the need for expensive fluorescent microscopy equipment and advanced training
Solution Approach 2:
The assay uses molecular tags that can be easily detected and do not require the expensive, complex fluorescent probes used in FISH. These tags are simpler, more cost-effective alternatives that maintain quantitative accuracy without requiring specialized equipment
3Measurement precision
If IHC is used to evaluate antigen expression, then expression levels are measured, but inter-laboratory variability cannot be completely prevented
Solution Approach 1:
The patent changes the detection parameter from antibody-based signal intensity (prone to variability) to molecular tag presence/absence or quantification. This parameter change makes the assay more robust and less susceptible to inter-laboratory variability in antibody performance and signal interpretation
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 accurate prediction of patient responsiveness to treatments binding CD137 and PD-L1, confirming the mode of action of therapeutic agents, and quantifying the expression and clustering of cell surface moieties, thereby optimizing treatment strategies.
Implementation Method 1
one of said first and second binding molecules comprises a molecular tag and the other of said first and second binding molecules comprises a cleavage-inducing moiety
Data Source
AI summary
The present disclosure relates to a method for detecting and/or quantifying expression of at least a first cell surface moiety and of a second cell surface moiety in a patient sample, and to a method for detecting and/or quantifying clustering of at least a first cell surface moiety with a second cell surface moiety in a sample, wherein the sample is exposed to a molecule having binding specificity for the at least first and second cell surface moieties. The present disclosure further relates to a method for predicting the responsiveness of a subject to such binding molecule, a method for determining the effectiveness of such binding molecule, a method for confirming the mode of action of such binding molecule, a method for treating a subject, and a method for screening one or more test agents for the ability to induce clustering of a first cell surface moiety with a second cell surface moiety.