Stable Complex Formation for Macromolecule Analysis
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Solution Overview
Problem
Current methods for analyzing macromolecules, such as proteins and peptides, face challenges including the need for high-affinity and specific binding agents, limited multiplexing capabilities, sensitivity, and stability in binding reactions, which affect the accuracy and efficiency of molecular recognition and characterization.
Innovation Solution
The method involves forming a stable complex between a binding agent and a target macromolecule using stabilizing components that are linked together, allowing for reversible binding and information transfer between nucleic acid tags, enabling enhanced specificity, stability, and multiplexing capabilities through the use of coding and recording tags, and adaptor molecules for encoding and decoding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If binding agents are used to recognize and bind target macromolecules, then molecular recognition and characterization can be performed, but the binding complexes lack sufficient stability for downstream processing
Solution Approach 1:
The patent introduces stabilizing components as intermediary molecules that mediate between the binding agent-target complex and the downstream processing steps. These stabilizing components form additional interactions with the complex, enhancing stability without requiring modification of the original binding agents or targets, thus resolving the contradiction between stability and complexity.
Solution Approach 2:
The patent creates composite binding complexes by combining binding agents, target macromolecules, and stabilizing components into a multi-component system. This composite structure leverages the strengths of each component: the binding agent provides specificity, the target provides the analyte, and the stabilizing components provide enhanced stability for downstream processing.
2Reliability
If crosslinking reagents are used to stabilize binding complexes, then complex stability improves, but reversibility and processing speed are reduced
Solution Approach 1:
The patent employs stabilizing components that can dynamically adjust their interaction strength with the binding complex. These components can form and break interactions reversibly, allowing the system to transition between stable (for processing) and labile (for release) states, thus resolving the contradiction between stability and processing speed.
Solution Approach 2:
The patent utilizes stabilizing components whose binding characteristics can be modulated by changing physical or chemical parameters such as temperature, pH, or ionic strength. This allows optimization of stability under processing conditions while enabling easy release by simply changing parameters, resolving the contradiction between stability and reversibility.
3Adaptability or versatility
If conventional immunoassay methods are used, then binding reactions can be performed, but multiplexing capabilities and sensitivity are limited
Solution Approach 1:
The patent segments the detection system into multiple independent binding agents, each specific to a different target macromolecule or epitope. Each binding agent-stabilizing component pair can be independently optimized and detected, enabling simultaneous multiplexed analysis while maintaining high sensitivity through the enhanced stability provided by the stabilizing components.
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 improves the stability and specificity of binding reactions, enabling higher temperature and longer duration encoding, reducing non-specific interactions, and facilitating efficient information transfer for macromolecule sequencing and analysis.
Implementation Method 1
allowing the binding agent to interact with a binding site located on the target
Implementation Method 2
linking the stabilizing components to form a stable complex comprising the binding agent, the target and the stabilizing components
Data Source
AI summary
The present disclosure relates to methods and kits for forming a stable complex comprising a binding agent and a target (e.g., a macromolecule). In some embodiments, the target comprises a peptide, a polypeptide, or a protein to be analyzed. In some embodiments, the present disclosure relates to formation of a stable complex comprising a binding agent and a target (e.g., a macromolecule) to be analyzed in a method which employs barcoding and nucleic acid encoding of molecular recognition events, and/or detectable labels. Provided herein is also a programmable system for information transfer comprising one or more adaptor molecules.