High-Resolution Cytometric Assays for Solid-Tumor Immune Profiling
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Solution Overview
Problem
Current cytometric assays struggle to provide high-resolution identification and isolation of immune cell populations in solid tumors, particularly in brain tumors, due to challenges in sample preparation and marker overlap, leading to ineffective therapies and lack of accurate prognostic information.
Innovation Solution
Development of cytometric panels with unique antibody combinations and improved sample preparation using Neutral Protease (Ch NP) for dissociating cells, enabling high-resolution identification and quantification of immune cell populations, including rare subsets, and providing kits for simultaneous characterization and isolation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multicolor FCM is used for high-throughput analysis of cells, then analysis speed and sensitivity are improved, but spectral overlap between fluorochromes and spreading error limit the number of markers that can be simultaneously stained
Solution Approach 1:
The patent changes the detection parameter from fluorescent light emission to light scattering properties. By using light scattering instead of fluorescence, the system avoids spectral overlap between markers while maintaining high-throughput analysis capability. This allows simultaneous staining for a large number of markers without the spectral interference that limits traditional multicolor FCM.
Solution Approach 2:
The patent replaces the optical detection system based on fluorescence with a light scattering-based detection system. This substitution eliminates the spectral overlap problem inherent in fluorescent detection while preserving the high-throughput flow cytometric analysis capability, enabling discrimination between multiple cell populations with many markers.
2Ease of operation
If flow cytometric sorting is used to isolate selected cell subsets, then cell isolation capability is improved, but the complexity of simultaneous staining for large number of markers increases due to spectral overlap and spreading error
Solution Approach 1:
The patent changes the detection parameter from fluorescence to light scattering, which fundamentally simplifies the staining complexity. Since light scattering does not suffer from spectral overlap or spreading error, the system can simultaneously stain for a large number of markers without increasing complexity, while maintaining the ability to isolate selected cell subsets based on their light scattering characteristics.
3Measurement precision
If CyTOF is used for phenotypic characterization with mass spectrometry, then spectral overlap limitation is overcome and high resolution characterization is achieved, but cell destruction occurs during the process preventing isolation and further examination
Solution Approach 1:
The patent replaces the mass spectrometry-based detection system with a light scattering-based flow cytometric system. This substitution maintains the high resolution characterization capability while preserving cell viability. The light scattering detection does not require cell destruction or ionization, allowing cells to remain intact for subsequent isolation and further examination after phenotypic characterization.
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
The assays enable accurate separation and characterization of immune cell populations in solid tumors, offering diagnostic and prognostic insights, and guiding personalized cancer therapies by correlating cell subsets with disease progression and survival.
Implementation Method 1
improved sample preparation using Neutral Protease (Ch NP) for dissociating cells
Implementation Method 2
a light source that activates any fluorescent antibodies that are bound to the flowing cells. An electronic detection apparatus collects the fluorescent light
Implementation Method 3
An electronic detection apparatus collects the fluorescent light that correlates with the amount of marker the cell is expressing
Data Source
AI summary
Provided relates to the field of cytometry, specifically to flow cytometric methods and kits for improved diagnosis, prognosis and monitoring of tumors and other lesions involving immune cell infiltration. Further provided are embodiments of the subject matter which relate to compositions and methods providing high resolution quantitative means for immunophenotyping and immune modeling, and for identification of disease prognostic and therapy predictive biomarkers.