Augurin Immunoassay Epitope Segmentation
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Solution Overview
Problem
Current assays for detecting augurin and its fragments or precursors lack high sensitivity, specificity, and reproducibility, with significant inter-assay variation, making them inadequate for detecting low concentrations effectively.
Innovation Solution
A sandwich immunoassay method utilizing specific binding of two antibodies, one produced by hybridoma cell line 482/H2 or its variants and the other by hybridoma cell line 439/F4 or its variants, targeting epitopes in the amino acid sequence spanning residues 71 to 107 of pre-augurin, allowing for the detection of augurin, argilin, Δ16-augurin, and Δ16-argilin with improved sensitivity and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional assays are used for detecting augurin, then the detection can be performed, but the sensitivity is insufficient and inter-assay variation is significant
Solution Approach 1:
The patent segments the detection process by using two different monoclonal antibodies that recognize distinct epitopes on augurin and its fragments. This segmentation allows the assay to detect multiple forms of augurin (full-length and C-terminal fragments) simultaneously, improving both sensitivity and reliability by reducing assay variation through multiple recognition points
Solution Approach 2:
The patent optimizes specific parameters including the selection of monoclonal antibodies with high affinity for different epitopes, optimization of incubation conditions, and selection of appropriate detection methods. These parameter changes enhance detection sensitivity while maintaining assay reliability by minimizing inter-assay variation
2Measurement precision
If conventional assays are used, then detection can be performed, but specificity for distinguishing augurin fragments is insufficient
Solution Approach 1:
The patent employs segmentation by using two distinct monoclonal antibodies targeting different epitopes on augurin. This approach enables specific detection of full-length augurin and its C-terminal fragments with high specificity, while the standardized sandwich assay format keeps the overall complexity manageable
Solution Approach 2:
The patent uses a sandwich assay format with two monoclonal antibodies as intermediaries that specifically bind to different epitopes on augurin. This intermediary approach enhances detection specificity by requiring dual recognition, while the established sandwich assay methodology maintains reasonable procedural complexity
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
The method achieves high sensitivity and specificity in detecting augurin and its fragments at low concentrations, with reduced inter-assay variation, enabling reliable detection and quantification in biological samples.
Implementation Method 1
augurin is detected by specific binding of a first and a second antibody
Data Source
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AI summary
The present invention relates to an immunoassay method for the detection of augurin or a precursor or fragment thereof comprising contacting a sample suspected of comprising augurin or a precursor or fragment thereof with a first and second antibody specific for augurin or a precursor or fragment thereof, wherein said first and second antibodies or antigen-binding fragments or derivatives thereof are specific for epitopes comprised in the sequence spanning amino acids 71 to 107 of pre-augurin according to SEQ ID NO:1.