CBX6 Demethylation Analysis for Memory B Cell Identification
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Solution Overview
Problem
Current methods for detecting and quantifying memory B cells are not specific and robust enough, particularly in complex samples, and often require cell purification and enrichment steps, which can be cumbersome and inefficient.
Innovation Solution
Analyzing the methylation status of specific CpG positions in the Chromobox protein homolog 6 (CBX6) gene region, where demethylation indicates memory B cells, allowing for their identification and quantification without purification or enrichment, using bisulfite convertibility and PCR-based methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If cell purification and enrichment steps are used to detect memory B cells, then measurement precision is improved, but device complexity and loss of time increase
Solution Approach 1:
The patent extracts the essential identifying feature of memory B cells (CBX6 demethylation status) from the complex cellular environment and uses it as a direct detection target. By focusing on this specific epigenetic marker rather than attempting to separate or enrich memory B cells from other cell types, the method achieves high measurement precision while eliminating complex purification workflows.
Solution Approach 2:
The patent uses DNA methylation status of the CBX6 gene as an intermediary marker to indirectly identify memory B cells. Instead of directly detecting memory B cells through complex phenotypic markers or physical separation, the method uses the epigenetic state of a specific gene region as a measurable proxy that uniquely identifies memory B cells in complex samples.
2Measurement precision
If cell purification and enrichment steps are used to detect memory B cells, then measurement precision is improved, but loss of time increases
Solution Approach 1:
The patent extracts the essential identifying feature of memory B cells (CBX6 demethylation status) from the complex cellular environment and uses it as a direct detection target. By focusing on this specific epigenetic marker rather than attempting to separate or enrich memory B cells from other cell types, the method achieves high measurement precision while eliminating complex purification workflows.
Solution Approach 2:
The patent performs preliminary characterization of memory B cells through epigenetic profiling to establish the CBX6 demethylation signature before actual detection. This preliminary understanding of the epigenetic landscape enables direct detection without time-consuming purification steps, as the demethylation status serves as an immediate identifying feature.
3Ease of operation
If conventional detection methods are used for memory B cells, then ease of operation is maintained, but measurement precision deteriorates in complex samples
Solution Approach 1:
The patent applies local quality by focusing detection on a specific local region (CBX6 gene promoter) with a specific epigenetic characteristic (demethylation status). Rather than attempting to detect memory B cells through their overall phenotypic properties in complex mixtures, the method targets this specific local molecular feature that provides unique identification, achieving high precision while maintaining operational simplicity.
Solution Approach 2:
The patent utilizes parameter changes in the epigenetic state of the CBX6 gene (methylation status) as a detection parameter. By measuring the methylation/demethylation state of this specific gene region, the method creates a clear distinguishable parameter that correlates with memory B cell identity, enabling precise detection in complex samples without complicating the detection process.
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 approach provides a reliable and specific method for distinguishing memory B cells from other cell types in complex samples, enabling accurate quantification and immune status assessment, applicable in various diagnostic and therapeutic contexts.
Implementation Method 1
The analyses according to the invention can identify memory B cells on an epigenetic level and distinguish them from all other cells in complex samples
Implementation Method 2
using bisulfite convertibility and PCR-based methods
Data Source
AI summary
The present invention relates to a method, in particular an in vitro method, for identifying memory B cells, comprising analyzing epigenetic modifications/properties of (including the methylation status) of at least one CpG position in the mammalian gene region for Chromobox protein homolog 6 (CBX6), wherein a demethylation or lack of methylation of said gene region is indicative for a memory B cell, when compared to a non-memory B cell. The analyses according to the invention can identify memory B cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The resent invention furthermore provides an improved method for quantifying memory B cells, in particular in complex samples. The method can be performed without a step of purifying and/or enriching cells, preferably in whole blood and/or non-trypsinized tissue.