Single-Cell Secretome Profiling for Multiplexed CAR T Evaluation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods lack a system and process for efficiently evaluating the safety and efficacy of cells for cellular therapies by analyzing the secretome of individual cells in a highly multiplexed manner, particularly for identifying suitable cells and assessing their response to target cells or stimulatory agents.
Innovation Solution
A method involving contacting subject cells with target cells or stimulatory agents, using a chamber system with an antibody panel to form antibody:secreted protein complexes, and imaging these complexes to identify the secretome, which includes peptides, polypeptides, proteins, and small molecules, allowing for the evaluation of cell safety and efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional methods are used to evaluate cellular therapy safety and efficacy, then the evaluation process is simpler, but the ability to simultaneously analyze thousands of single cells from heterogeneous populations is insufficient
Solution Approach 1:
The system divides the heterogeneous cell population into individual single cells for separate analysis in microchambers, and divides the detection process into multiple sequential steps (secretion collection, antibody binding, imaging) that can be performed in a standardized workflow across thousands of cells simultaneously
Solution Approach 2:
The microchamber array system serves multiple functions: it acts as both the reaction vessel for cell stimulation and secretion collection, and as the platform for multiplexed antibody binding and imaging detection. The same chamber structure supports both biological function and analytical detection
2Measurement precision
If highly multiplexed analysis is implemented to simultaneously analyze thousands of single cells, then the measurement precision and functional heterogeneity detection improve, but the device complexity and operational difficulty increase
Solution Approach 1:
The system performs preliminary actions by pre-coating microchambers with specific antibodies or capture reagents before cell loading, and by pre-establishing the microchamber array structure optimized for both cell culture and detection. This preliminary preparation standardizes the workflow and reduces operational complexity during actual analysis
Solution Approach 2:
The system introduces intermediary elements such as antibody-coated microchambers that mediate between the secreted proteins from cells and the detection system. These intermediaries capture and concentrate secretome components, enabling precise measurement while simplifying the detection workflow
3Loss of information
If conventional secretome analysis methods are used, then the operational procedure is simpler, but the ability to detect functional heterogeneity among individual cells is insufficient
Solution Approach 1:
The system segments the cell population analysis into individual single-cell measurements, with each microchamber containing and analyzing one cell's secretome. This segmentation preserves all functional information from each individual cell without averaging effects that would mask heterogeneity
Solution Approach 2:
The system creates multiple identical copies of the analysis platform (thousands of microchambers with identical structure and reagents) to simultaneously capture and analyze secretomes from many individual cells. This copying approach enables comprehensive sampling of cellular heterogeneity while maintaining standardized detection conditions
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 the identification of safe and efficacious cellular therapies by analyzing the secretome of cells, including T-lymphocytes expressing CARs, to determine their response to target cells, ensuring selective and potent immune activation without autoimmune or inflammatory responses.
Implementation Method 1
the subject cell to secrete at least one of a peptide, polypeptide, and protein
Implementation Method 2
at least one antibody of the antibody panel specific for the at least one protein to bind the at least one peptide, polypeptide, or protein, forming at least one of an antibody: secreted peptide, antibody: secreted polypeptide, or an antibody: secreted protein complex
Implementation Method 3
imaging the at least one peptide, polypeptide, or protein, forming at least one of an antibody:secreted peptide, antibody:secreted polypeptide, or an antibody: secreted protein complex
Data Source
Figure 1
Figure 2
Figure 3
AI summary
Disclosed are methods of identifying a secretome from a subject cell within a heterogeneous cell population when the subject cell contacts a target cell (e.g. a subject immune cell contacts a target cancer cell) or a stimulatory agent and methods of using the identified secretome to identify cells that are safe and efficacious for cellular therapies, including adoptive CAR T-cell therapies.