Methylated DNA Detection via Polypeptide Coating
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Solution Overview
Problem
Current methods for detecting methylated DNA, such as those using methylation-sensitive restriction enzymes or bisulfite treatment, are limited in their ability to analyze CpG methylation patterns across the genome and struggle with high-throughput applications, especially when dealing with low DNA amounts.
Innovation Solution
A single-tube assay method involving a polypeptide capable of binding methylated DNA, coated onto a container, which allows for direct detection and analysis of methylated DNA without the need for bisulfite treatment or methylation-sensitive restriction enzymes, enabling the estimation of methylation density and detection of methylated DNA even in small amounts.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If methylation-sensitive restriction enzymes or bisulfite treatment are used for detecting methylated DNA, then detection capability is achieved, but the methods are limited in high-throughput applications and struggle with low DNA amounts
Solution Approach 1:
The patent extracts the methylated DNA from the complex genomic background by using antibodies specific to 5-methylcytosine that selectively bind to methylated DNA sequences. This extraction approach allows the method to focus on only the relevant methylated portions of DNA, enabling high-throughput analysis without being overwhelmed by the complexity of the entire genome, thereby resolving the contradiction between detection reliability and productivity.
2Reliability
If methylation-sensitive restriction enzymes or bisulfite treatment are used, then detection is possible, but the ability to analyze CpG methylation patterns across the genome is limited
Solution Approach 1:
The patent employs a universal antibody-based detection system that can recognize 5-methylcytosine regardless of its genomic context. The same antibody reagents can be used to analyze methylation patterns at any CpG site across the entire genome, making the method universally applicable for genome-wide methylation analysis while maintaining detection accuracy, thus resolving the contradiction between reliability and adaptability.
3Reliability
If traditional methods are used for methylated DNA detection, then detection can be performed, but sensitivity is reduced when dealing with low DNA amounts
Solution Approach 1:
The patent introduces antibodies as intermediary molecules that specifically bind to 5-methylcytosine residues on methylated DNA. These antibodies act as mediators that amplify the detection signal, allowing even small amounts of methylated DNA to be detected with high reliability. The antibody-DNA complex can then be detected through various downstream methods, enabling sensitive detection in low DNA amount scenarios while maintaining detection reliability.
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 provides a fast, sensitive, and reliable detection of methylated DNA, capable of analyzing methylation degrees and generating methylation profiles of single or multiple gene loci, suitable for high-throughput applications and detection in limited samples.
Implementation Method 1
a polypeptide capable of binding methylated DNA
Data Source
AI summary
The present invention relates to an in vitro method for detecting methylated DNA comprising (a) coating a container with a polypeptide capable of binding methylated DNA; (b) contacting said polypeptide with a sample comprising methylated and/or unmethylated DNA; and (c) detecting the binding of said polypeptide to methylated DNA. In a preferred embodiment, said method further comprises step (d) analyzing the detected methylated DNA by sequencing. Another aspect of the present invention is a kit for detecting methylated DNA according to the methods of the invention comprising (a) a polypeptide capable of binding methylated DNA; (b) a container which can be coated with said polypeptide; (c) means for coating said container; and (d) means for detecting methylated DNA.