Antigen-Specific IgE Quantification Using Mouse Serum Standards
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Solution Overview
Problem
Current methods for quantifying antigen-specific IgE in dogs and humans are limited by issues with unit standardization, antigen-specific IgE purification, hybridoma preparation, and high costs, making absolute quantitation and comparison between different antigens impossible.
Innovation Solution
A method using rat monoclonal antibodies that recognize both canine and mouse IgE to generate standard curves for quantifying antigen-specific IgE, allowing for the use of sensitized mouse sera to standardize measurements across different antigens, and a kit containing antibodies and antigens for convenient quantitation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a standard serum is prepared using an experimental dog for each antigen, then antigen-specific IgE can be quantified, but the preparation cost becomes extremely high (160,000 to 260,000 yen per antigen)
Solution Approach 1:
The patent uses mouse serum as a copy substitute for expensive dog standard serum. Mouse serum is much cheaper to prepare and obtain, yet it can still be used to generate standard curves for quantitating antigen-specific IgE in dogs through cross-reactivity of the detection antibodies, thereby dramatically reducing costs while maintaining measurement precision
Solution Approach 2:
The patent replaces expensive, long-term investment in experimental dog preparation with inexpensive, easily obtainable mouse serum that can be prepared quickly and at low cost. This disposable approach eliminates the need for expensive long-term animal maintenance and breeding programs
2Measurement precision
If sensitized dog serum is used as standard serum, then absolute quantitation of antigen-specific IgE is achieved, but ethical concerns arise and preparation is time-consuming
Solution Approach 1:
The patent creates a copy of the standard serum using mouse instead of dog. Mouse serum can be prepared much faster and with fewer ethical constraints, while still serving the functional purpose of generating standard curves for absolute quantitation through the cross-reactive detection system
3Measurement precision
If different standard sera are used for different antigens in the same laboratory, then antigen-specific IgE can be measured, but the unit of indication cannot be unified across different antigens
Solution Approach 1:
The patent develops a universal standard curve system using mouse serum that can be applied across different antigens. The same mouse-based standard curve generation method works for multiple antigens, allowing unified unit of indication (IU/ml) to be used consistently across all antigen-specific IgE measurements in the laboratory
4Measurement precision
If purification of antigen-specific IgE is performed using two-stage purification, then antigen-specific IgE can be isolated, but the process becomes complex and elution success is uncertain due to strong IgE-antigen binding
Solution Approach 1:
The patent extracts and removes the need for complex two-stage purification processes. Instead of attempting to purify antigen-specific IgE from dog serum through complex chromatography, the patent directly uses mouse serum as the standard, eliminating the purification step entirely while maintaining measurement precision
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 comparative quantitation of antigen-specific IgE, facilitating the monitoring of allergic diseases and therapeutic effects, reducing costs and ethical concerns by using mouse models.
Implementation Method 1
using a substance that recognizes canine IgE and also recognizes the IgE of the laboratory animal
Implementation Method 2
contacting a biological sample from a subject dog with an antigen and allowing the IgE in the sample to bind the antigen
Data Source
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AI summary
Provided are convenient and effective means of quantifying antigen-specific canine and human IgEs, specifically a method of quantifying antigen-specific canine IgE and a method of quantifying antigen-specific human IgE, each comprising the following steps (A) to (D): (A) a step for contacting a biological sample from a subject with an antigen to bind the IgE in the sample and the antigen, (B) a step for measuring the IgE in a standard sample from a laboratory animal other than the subject, sensitized with the antigen of the step (A), using a substance that recognizes subject IgE and also recognizes the IgE of the laboratory animal, and generating a standard curve, (C) a step for detecting the conjugate formed in the step (A), using a substance that recognizes subject IgE and also recognizes the IgE of the laboratory animal, used in the step (B), and (D) a step for quantifying the amount of IgE in the biological sample from the subject using the amount of conjugate detected in the step (C) and the standard curve generated in the step (B).