mRNA Expression Quantitation With Common Peptide Labels
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Solution Overview
Problem
Existing methods for evaluating mRNA vaccine efficacy face challenges due to the lack of high-specificity antibodies, difficulty in distinguishing between exogenous and endogenous proteins, and limited versatility in measuring multiple protein expression levels, leading to inaccurate and time-consuming assessments.
Innovation Solution
A method using fusion mRNAs with linked labeling nucleotides encoding a common peptide, allowing expression and quantitation with a single antibody to accurately measure and compare the expression levels of multiple mRNAs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional immunological assays with specific antibodies are used to measure protein expression levels, then measurement accuracy can be achieved, but the process becomes time-consuming and costly due to the need to obtain high-specificity antibodies for each target protein
Solution Approach 1:
The patent introduces a labeling peptide as an intermediary element that is fused to the target protein. Instead of requiring specific antibodies for each target protein, a common antibody against the labeling peptide is used to detect all target proteins. This intermediary approach resolves the contradiction by enabling accurate measurement without the time-consuming process of obtaining multiple specific antibodies
Solution Approach 2:
The labeling peptide serves as a universal tag that can be attached to multiple different target proteins. A single common antibody can recognize this universal tag across different target proteins, making the measurement system universally applicable to various proteins without requiring protein-specific antibodies for each target
2Measurement precision
If antibodies are used to detect target proteins, then protein expression can be measured, but the ability to distinguish between endogenous and exogenous target proteins is lost
Solution Approach 1:
The target protein is segmented into two functional parts: the original target protein domain and the attached labeling peptide domain. The labeling peptide is an artificial addition that does not exist in the endogenous protein, creating a detectable difference between endogenous and exogenous proteins while maintaining the target protein's functional properties
Solution Approach 2:
The labeling peptide acts as an intermediary marker that provides information about the exogenous nature of the protein. The common antibody against the labeling peptide specifically detects this marker, thereby distinguishing exogenous labeled proteins from endogenous unlabeled proteins in the same sample
3Measurement precision
If multiple specific antibodies are used to measure different target proteins, then individual protein expression levels can be determined, but the versatility of the measurement system decreases
Solution Approach 1:
The labeling peptide and common antibody system creates a universal measurement platform that can be applied to multiple different target proteins. By attaching the same labeling peptide to different target proteins of interest, the same common antibody can detect all of them, making the system highly versatile and adaptable to various proteins without requiring new antibodies for each target
Solution Approach 2:
The system changes the measurement parameter from target-protein-specific antibody binding to universal labeling-peptide-specific antibody binding. This parameter change enables the measurement system to handle multiple different target proteins uniformly, increasing versatility while maintaining the ability to measure individual protein expression levels through quantitation of the labeled proteins
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
Enables accurate, versatile, and efficient quantitation of mRNA expression levels using a common antibody, simplifying the evaluation process and overcoming the limitations of existing methods.
Implementation Method 1
a quantitation step of quantitating, using the same antibody against the labeling peptide, a labeled polypeptide expressed from each labeled mRNA
Data Source
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AI summary
A problem to be addressed by the present invention is to develop a method capable of immunologically simply and accurately quantitate and analyze proteins in order that the function and effect of two or more mRNAs having different sequences can be compared and evaluated in the form of the expression levels of the proteins encoded by the mRNAs. Labeled mRNAs in each of which a labeling nucleotide encoding the same labeling peptide is linked to an end of an mRNA are prepared. The respective expression levels of the labeled polypeptides expressed from the labeled mRNAs are quantitated using the same antibody against the labeling peptide.