Erythroid Cell Hemoglobin Quantification by Fluorescent Flow Cytometry
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Solution Overview
Problem
Current methods for determining the hemoglobin content, particularly HbF, in each red blood cell are not sensitive enough and only provide qualitative measurements, limiting their ability to explain clinical response variabilities and anticipate treatment adjustments in diseases like sickle cell disease.
Innovation Solution
A method involving the isolation of erythroid cells, permeabilization of their membranes, labeling with anti-hemoglobin antibodies conjugated to fluorochromes, and measurement by flow cytometry to accurately determine the hemoglobin content in each erythroid cell, allowing for precise quantification and distribution analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to determine hemoglobin content in red blood cells, then the measurement process is simple, but the measurement precision and sensitivity are insufficient
Solution Approach 1:
The patent uses fluorescently labeled antibodies as intermediaries to detect hemoglobin content. The antibodies specifically bind to hemoglobin molecules, and the fluorescent labels amplify the detection signal, enabling precise quantification of hemoglobin content in individual red blood cells through flow cytometry.
Solution Approach 2:
The patent replaces conventional mechanical or chemical hemoglobin measurement methods with a fluorescence-based optical detection system. By using fluorochrome-conjugated antibodies and flow cytometry, the method substitutes traditional measurement mechanics with optical signal detection, achieving superior sensitivity and precision.
2Loss of information
If conventional hemoglobin measurement methods are used, then the procedure is straightforward, but the ability to explain clinical response variabilities is limited
Solution Approach 1:
The patent enables precise measurement of hemoglobin content in individual red blood cells, providing detailed information about hemoglobin distribution and concentration variations. This granular data serves as feedback to explain clinical response variabilities and guide personalized treatment adjustments for conditions like sickle cell disease.
Solution Approach 2:
The patent measures multiple parameters including hemoglobin content, fluorescence intensity, and cell characteristics simultaneously. By analyzing variations in these parameters across individual cells, the method captures clinical response information that conventional bulk measurement methods miss, enabling better understanding of disease mechanisms and treatment responses.
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
This method enables accurate and sensitive determination of hemoglobin content in each red blood cell, facilitating better understanding of clinical responses and enabling more tailored treatment approaches for conditions such as sickle cell disease.
Implementation Method 1
labeling the at least one hemoglobin Hbx of the erythroid cells obtained in step b) with at least one anti-Hbx antibody conjugated to a fluorochrome capable of emitting a fluorescence
Data Source
AI summary
The invention concerns a method for determining, by flow cytometry, the hemoglobin content of each erythroid cell of a set of erythroid cells. This method applies in particular to determining the hemoglobin content of each red blood cell of a set of red blood cells. The invention also concerns a method for determining the amount of red blood cells transfused into a patient and for monitoring the therapeutic efficacy of a treatment for sickle cell disease or β-thalassemia.