Pediatric Tumor Flow Cytometry Antibody Panel
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Solution Overview
Problem
Current diagnostic methods for pediatric solid tumors are challenging due to similarities in morphological features, leading to misdiagnosis and the lack of a uniformly recommended panel of antibodies for sensitive and specific diagnosis and classification, with multiparameter flow cytometry not being routinely used for solid tumors despite its potential.
Innovation Solution
A novel ≥8-color/fluorochrome antibody combination for simultaneous detection of ≥12 different proteins, allowing for the identification and classification of pediatric cancer and detailed analysis of tumor-infiltrating immune cells using a kit comprising fluorochrome-conjugated antibodies against specific markers, enabling accurate diagnosis and classification of pediatric solid tumors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional morphological and immunohistochemical analysis is used for pediatric solid tumors, then diagnosis can be performed with basic equipment, but misdiagnosis occurs due to similar morphological features of small round cell tumors
Solution Approach 1:
The patent combines multiple immunohistochemical markers (CD45, CD56, CD99, GD2, EpCAM, CD271, and nuclear markers) into a single integrated diagnostic panel that can be applied to tissue sections. This merging of multiple diagnostic criteria into one unified approach allows simultaneous assessment of tumor cells and infiltrating immune cells, resolving the contradiction between diagnostic accuracy and procedural complexity.
Solution Approach 2:
The diagnostic method is designed to be universally applicable across multiple pediatric solid tumor types including neuroblastoma, Ewing sarcoma, rhabdomyosarcoma, and Wilms tumor. The same panel of markers serves multiple functions: identifying tumor cells, classifying tumor types, and characterizing the tumor microenvironment, thereby improving diagnostic accuracy without proportionally increasing complexity.
2Measurement precision
If a limited number of markers is used in flow cytometry for pediatric solid tumors, then the procedure is simpler, but it cannot accurately differentiate among different tumor types
Solution Approach 1:
The patent segments the diagnostic information into distinct functional groups: tumor cell identification markers (CD45-, CD56+), tumor type-specific markers (CD99 for Ewing sarcoma, GD2 for neuroblastoma, EpCAM for Wilms tumor, nuclear markers for rhabdomyosarcoma), and immune cell characterization markers. This segmentation allows the complex panel to be systematically organized and applied, managing the complexity while maintaining high classification accuracy.
Solution Approach 2:
The patent changes the parameters of flow cytometry analysis by expanding from traditional limited marker panels to a comprehensive multi-parameter panel including surface markers, cytoplasmic markers, and nuclear markers. This parameter expansion enables accurate differentiation of tumor types while the use of standardized flow cytometry technology keeps the procedural complexity manageable.
3Loss of information
If multiparameter flow cytometry is applied to pediatric solid tumors, then simultaneous characterization of tumor cells and infiltrating immune cells is achieved, but the diagnostic workflow becomes more complex
Solution Approach 1:
The patent applies preliminary action by performing immunohistochemical staining of tissue sections with multiple markers before flow cytometry analysis. This preliminary staining of fixed tissue sections preserves the spatial context and allows subsequent flow cytometry to focus on quantitative analysis, thereby capturing comprehensive microenvironment information while managing analytical complexity through staged processing.
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 solution enables accurate identification and classification of pediatric cancer into different diagnostic entities and simultaneous assessment of tumor-infiltrating immune cells, providing critical information for diagnosis, monitoring, and selecting appropriate therapies.
Implementation Method 1
A novel ≥8-color/fluorochrome antibody combination for simultaneous detection of ≥12 different proteins
Data Source
AI summary
A kit-of-parts for the flow cytometric detection of pediatric tumor cells, comprising fluorochrome-conjugated antibodies directed against the cell surface markers CD45, CD56, GD2, CD99, CD8, EpCAM, CD4, smCD3, CD19 and CD271, the cytoplasmic marker cyCD3, and the nuclear marker(s) nuMyogenin and/or nuMyoD1, wherein(i) the antibodies against the markers CD99/CD8 are conjugated to the same fluorochrome and representing a first marker pair CD99/CD8;(ii) the antibodies against the markers EpCAM/CD4 are conjugated to the same fluorochrome and representing a second marker pair EpCAM/CD4;(ii) the antibody against CD271 is conjugated to the same fluorochrome as the antibody against either cyCD3 or smCD3 and representing a third marker pair CD271/cyCD3 or CD271/smCD3;wherein between the first, second and third marker pairs the fluorochromes are distinguishable; and wherein the antibodies against the cytoplasmic and the nuclear markers are physically separated from the antibodies against the cell surface markers.