Multivalent Affinity Reagents for Single-Molecule Signal Detection
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Solution Overview
Problem
Existing affinity reagents lack the ability to produce strong and reliable signals for single molecule detection and accurate quantification, and do not bind avidly enough to target molecules for reliable detection at single molecule resolution.
Innovation Solution
The development of affinity reagents comprising a retaining component with one or more label components and binding components, where the detectable probe has a lower equilibrium dissociation constant and dissociation rate constant than individual binding components, allowing for enhanced binding and detection characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional affinity reagents are used, then the structure and properties of biomolecules can be characterized, but the signal strength is insufficient for single molecule detection and accurate quantification
Solution Approach 1:
The patent combines multiple binding components (such as multiple antibody binding sites) onto a single retaining component structure. This merging of multiple binding functions into one unified probe structure enables the affinity reagent to bind with higher overall avidity to target molecules, producing sufficiently strong signals for single molecule detection while maintaining measurement reliability for accurate quantification.
Solution Approach 2:
The affinity reagent is constructed as a composite structure comprising a retaining component (such as a nanoparticle or protein scaffold) that holds multiple binding components (antibodies, antigen-binding fragments). This composite architecture allows the probe to achieve both high binding avidity for reliable single molecule detection and sufficient signal strength for accurate quantification, resolving the contradiction between detection sensitivity and measurement reliability.
2Reliability
If traditional affinity reagents are used, then binding to target molecules can occur, but the binding avidity is insufficient for detection at single molecule resolution
Solution Approach 1:
Multiple binding components are merged onto a single retaining component, creating a multi-valent affinity reagent. This merging approach increases the overall binding avidity to target molecules at the single molecule level, enabling reliable detection while maintaining the ability to perform accurate quantification through the retained signal strength.
Data Source
AI summary
An affinity reagent, having: (a) a retaining component such as a structured nucleic acid particle; and (b) one or both of (i) one or more label components attached to the retaining component, and (ii) one or more binding components attached to the retaining component.