Multicolor Antibody Panels for PID Flow Cytometry Reproducibility

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Solution Overview

Problem

Current flow cytometric protocols for diagnosing primary immunodeficiencies (PID) face challenges in reproducibility and data exchange between centers due to differences in antibody panels, sample handling, and instrument setup, failing to provide a comprehensive overview of lymphocyte subsets and immunoglobulin isotypes, particularly in B-cells, and are not standardized for multicenter settings.

Innovation Solution

Development of optimized reagent compositions comprising specific combinations of fluorochrome-conjugated antibodies in multicolor tubes for flow cytometric immunophenotyping, designed to identify and dissect maturation and functional subsets of lymphocytes, including B-cells and T-cells, with distinct fluorochrome combinations for enhanced brightness, compensation, and stability, and evaluated in multicenter studies for reproducibility and efficiency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If multiple different antibody panels are used by different centers for PID diagnosis, then each center can optimize for their specific needs, but reproducibility and data exchange between centers deteriorate

Engineering Contradiction:

Improvecenter-specific optimizationVSAvoidreproducibility between centers

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent establishes a universal reference framework that can be applied across all diagnostic centers. The reference profiles and standardized protocols serve as a common platform that enables data exchange and comparison between centers while allowing each center to maintain their own optimized protocols within the standardized framework.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Loss of information

If comprehensive immunophenotypic panels are used to identify multiple subpopulations, then detailed description of lymphocyte subsets is improved, but device complexity and protocol difficulty worsen

Engineering Contradiction:

Improvecomprehensive cell subset informationVSAvoidprotocol complexity

Core Design Contradiction:

Loss of informationVSDevice complexity

Solution Approach 1:

The patent segments the comprehensive immunophenotyping process into distinct functional modules: reference profile generation, sample analysis, and automated comparison. This segmentation allows each module to be optimized independently while maintaining overall system comprehensiveness, reducing the perceived complexity for users.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent creates reference profiles that serve as templates or copies of normal immunophenotypic patterns. These reference profiles can be stored and reused across multiple analyses, eliminating the need to re-establish comprehensive cell subset definitions for each new sample, thereby reducing protocol complexity while maintaining comprehensive information capture.

Inventive Principle:

Principle #26Copying

3Productivity

If disease-specific antibody panels are used for targeted screening, then diagnostic efficiency for specific conditions is improved, but comprehensive overview of all lymphocyte subsets deteriorates

Engineering Contradiction:

Improvediagnostic efficiencyVSAvoidoverall immune system overview

Core Design Contradiction:

ProductivityVSLoss of information

Solution Approach 1:

The patent performs preliminary comprehensive analysis by generating reference profiles that encompass all lymphocyte subsets before focusing on disease-specific markers. This preliminary action ensures that comprehensive immune system information is captured first, allowing subsequent disease-specific analysis to build upon this complete baseline rather than working from a limited perspective.

Inventive Principle:

Principle #10Preliminary action

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 proposed antibody panels enable unequivocal identification and classification of lymphocyte subsets, improving reproducibility and providing a detailed description of immune cell compartments, facilitating accurate diagnosis and classification of PID patients across different centers, and are adaptable for immune monitoring and therapy.

Implementation Method 1

fluorochrome-conjugated antibodies in multicolor tubes for flow cytometric immunophenotyping

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Implementation Method 2

specific combinations of fluorochrome-conjugated antibodies designed to identify and dissect maturation and functional subsets of lymphocytes

Methodology Applied

Scientific EffectAntibody-antigen binding:

Data Source

PatentUS20240345085A1Reagents, methods and kits for diagnosing primary immunodeficiencies

Publication Date: 2024.10.17 ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC

AI summary

This invention relates to the field of primary immunodeficiencies (PID), more specifically to means and method for the diagnosis of PID of the lymphoid system. Provided are unique reagent compositions for the flow cytometric immunophenotyping of leukocytes comprising fluorochrome-conjugated antibodies directed against various specific combinations of markers. Also provided are kits comprising the reagent compositions, and methods using the same.