Immunological mRNA Expression Evaluation With Shared Peptide Tags
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Solution Overview
Problem
Existing methods for evaluating mRNA vaccine efficacy face challenges due to the lack of specific 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 labeling nucleotides encoding a common peptide, allowing expression and quantitation with a single antibody, enabling accurate and versatile evaluation of mRNA expression levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a conventional immunological assay with a specific antibody is used to measure protein expression level, then the measurement can be performed, but it requires obtaining a highly specific antibody which involves large amount of time and cost
Solution Approach 1:
The patent introduces a labeling peptide as an intermediary element. Instead of directly measuring the target protein with a specific antibody, the labeling peptide serves as a mediator that is selectively expressed only when the target mRNA is present. The antibody against the labeling peptide indirectly detects the target protein expression, eliminating the need to find or produce a highly specific antibody for the target protein itself.
Solution Approach 2:
The labeling peptide acts as a copy or proxy for the target protein. Rather than directly interacting with the target protein, the system creates a simplified copy (labeling peptide) that carries the information about target protein expression. This copy can be easily detected by a common antibody, replacing the complex task of finding a highly specific antibody for the target protein.
2Measurement precision
If an antibody with high specificity is used to measure target protein, then measurement accuracy improves, but the antibody cannot distinguish between exogenous target protein from mRNA and endogenous target protein
Solution Approach 1:
The patent segments the protein detection into two distinct components: the target protein (which may be endogenous or exogenous) and the labeling peptide (which is exclusively expressed from the introduced mRNA). By separating these functions, the system can detect the labeling peptide's presence to determine mRNA expression, while the target protein's presence alone cannot be confused with endogenous protein since the labeling peptide provides the specific marker.
Solution Approach 2:
The labeling peptide serves as an intermediary marker that bridges the gap between mRNA introduction and protein detection. It mediates the distinction by being selectively expressed only when the target mRNA is introduced, allowing the antibody to detect this intermediary rather than directly detecting the target protein, thus avoiding the confusion between exogenous and endogenous proteins.
3Adaptability or versatility
If multiple antibodies with high specificity are provided to measure multiple target proteins, then measurement versatility improves, but the complexity and cost increase significantly
Solution Approach 1:
The patent creates a universal detection system where a single antibody against the labeling peptide can detect the expression of multiple different target proteins. Each target mRNA is designed to express the same labeling peptide, so one antibody serves multiple functions by detecting the labeling peptide as a common marker across different target proteins, eliminating the need for multiple specific antibodies.
Solution Approach 2:
The patent merges the detection function for multiple target proteins into a single detection mechanism. Instead of requiring separate antibodies for each target protein, the system combines them by using the labeling peptide as a common marker. The antibody against the labeling peptide merges the detection of multiple proteins into one unified measurement approach.
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 and efficient quantitation and comparison of protein expression levels from multiple mRNAs with different sequences using a common antibody, overcoming the limitations of existing methods.
Implementation Method 1
A method for immunological evaluation of an mRNA expression level, comprising: an expression step of allowing labeled mRNAs to perform expression... a labeling nucleotide encoding the same labeling peptide, linked to an end of each target RNA
Implementation Method 2
a quantitation step of quantitating, using the same antibody against the labeling peptide, a labeled polypeptide expressed from each labeled mRNA in the expression step
Data Source
AI summary
A method is capable of immunologically simply and accurately quantitating and analyzing proteins that the function and effect of two or more mRNAs having different sequences are configured to 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.