Single-Cell Multiplexed Analysis Using Repeating Capture Agent Patterns
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods lack the capability for high-throughput, multiplexed analysis of intracellular protein interactions and signaling within a single cell, particularly in capturing and analyzing hundreds of intracellular components across thousands of cells in parallel.
Innovation Solution
The system employs a composition with a repeating pattern of capture agents that disrupts cell membranes upon contact, exposing intracellular components to these agents for complex formation, allowing for the visualization of protein expression, regulation, and signaling events within a controlled release environment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional single-cell analysis methods are used, then detailed molecular profiling can be achieved, but the throughput is limited and cannot analyze hundreds of components across thousands of cells in parallel
Solution Approach 1:
The system divides the analysis into discrete spatial units (chambers) and molecular components (capture agents for different proteins/DNA/RNA). Each chamber can independently analyze one or more cells for multiple components simultaneously, enabling parallel processing across thousands of cells while maintaining detailed molecular profiling through the capture agent library
Solution Approach 2:
The capture agents are designed to be multi-functional, capable of binding to different intracellular components (proteins, DNA, RNA) within the same cell. The universal substrate design with standardized capture agent integration allows a single platform to analyze multiple molecular types across thousands of cells simultaneously, achieving high throughput without proportionally increasing system complexity
2Quantity of substance
If multiplexed analysis of intracellular components is implemented, then comprehensive profiling of hundreds of components is enabled, but the ability to maintain single-cell resolution and analyze components in parallel is compromised
Solution Approach 1:
The system segments the analysis space into discrete chambers, each capable of containing and analyzing individual cells. This spatial segmentation maintains single-cell resolution while enabling parallel analysis of multiple cells across different chambers. The capture agents within each chamber specifically bind to intracellular components, allowing comprehensive profiling of hundreds of components while preserving the ability to track each component's expression at the single-cell level
Solution Approach 2:
Each chamber is designed with specific local conditions (capture agent composition, cell lysis conditions) optimized for analyzing particular intracellular components. This local quality approach allows comprehensive multiplexed analysis while maintaining the precision needed for single-cell resolution, as each chamber's microenvironment is tailored to preserve the integrity and detectability of the specific components being analyzed in that particular cell
3Reliability
If cell lysis is performed to expose intracellular components, then protein expression and interactions can be analyzed, but the membrane disruption may cause loss of intracellular components or contamination
Solution Approach 1:
The system performs preliminary cell lysis within the controlled environment of the chamber before capture agent binding. This preliminary action ensures that intracellular components are released and stabilized in a controlled manner, preventing contamination from external sources. The lysis conditions are optimized to maintain component integrity while ensuring complete exposure of intracellular targets for accurate capture and analysis
Solution Approach 2:
The chamber structure serves as an intermediary between the cell lysis process and the capture agent binding process. It provides a controlled microenvironment that facilitates controlled lysis while preventing unwanted contamination. The chamber acts as a mediator that allows membrane disruption to occur in a controlled manner, ensuring that intracellular components are released and stabilized before capture agents bind, thereby maintaining reliability of component capture accuracy while minimizing harmful artifacts from uncontrolled membrane disruption
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 simultaneous analysis of hundreds of intracellular components across thousands of cells, providing detailed insights into cellular processes and signaling pathways, overcoming the limitations of existing technologies by facilitating efficient and scalable multiplexed analysis.
Implementation Method 1
a coating composition comprising a cell lysis composition
Implementation Method 2
Capture agents of the disclosure specifically bind at least one intracellular component of the cell to form at least one complex
Data Source
AI summary
Disclosed are compositions and methods for the multiplexed analysis of one or more intracellular targets of a single cell. Exemplary compositions of the disclosure comprise a surface comprising a plurality of capture agents operatively-linked thereto, wherein each capture agent specifically binds to a distinct intracellular target and wherein the plurality of capture agents form a repeating pattern; a substrate comprising a plurality of chambers, wherein the substrate releasably couples with the surface and wherein each chamber of the plurality of chambers comprises at least one repeat of the repeating pattern of the plurality of capture agents of the surface; a coating composition comprising a cell lysis composition; and a linker composition comprising a functionalization component and an extension component.