Native IgE Detection via FcεRlα Receptor Binding
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Solution Overview
Problem
Current methods for identifying conformational epitopes of allergens, especially new and unknown ones, are complex, costly, and often destroy the native structure of proteins, making it difficult to achieve a native interaction between allergen-specific IgE and allergens for effective detection and diagnosis of parasitic or fungal infections.
Innovation Solution
A method using the high-affinity IgE receptor FcεRlα-(His)6 to bind allergen-specific IgE to a solid phase surface, maintaining its native conformation, allowing for the selective binding and detection of allergens through their conformational epitopes, utilizing a recombinant manufacturing process and histidine tags to keep IgE at a sufficient distance from the surface.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods (SDS-PAGE, 2D gel electrophoresis) are used to identify allergens, then linear epitopes can be detected, but conformational epitopes are destroyed due to denaturing techniques
Solution Approach 1:
The patent changes the fundamental parameter of the detection environment from denaturing to native conditions. By using a native gel electrophoresis system instead of SDS-PAGE or 2D gel electrophoresis, the three-dimensional structure of allergens is preserved, allowing conformational epitopes to remain intact and detectable by IgE antibodies.
Solution Approach 2:
The patent creates a copy of the native interaction environment by using recombinant allergens produced under conditions that maintain their natural conformation. These recombinant proteins serve as faithful copies of native allergens, preserving conformational epitopes while enabling controlled experimental conditions.
2Measurement precision
If phage display libraries are used to identify conformational epitopes, then binding peptides can be selected, but the process is complex and time-consuming
Solution Approach 1:
The patent extracts the essential function of phage display (selecting binding molecules) and simplifies it by directly using IgE antibodies from patient sera without requiring phage construction, library generation, or iterative biopanning steps. This extraction of the core binding selection function eliminates the complex phage display apparatus while achieving the same epitope identification goal.
Solution Approach 2:
Instead of creating and maintaining complex phage display libraries that require extensive infrastructure and multiple screening rounds, the patent uses readily available patient sera containing natural IgE antibodies. These disposable biological samples eliminate the need for expensive, time-consuming phage library construction and maintenance.
3Quantity of substance
If affinity chromatography is used to purify specific IgE, then pure IgE can be obtained, but the procedure is complex and requires known allergens
Solution Approach 1:
The patent makes the detection system universal by using total IgE from patient sera directly without purification. The native gel electrophoresis system can detect all IgE subclasses and specificities simultaneously, eliminating the need for allergen-specific affinity chromatography steps that require prior knowledge of the target allergen.
Solution Approach 2:
The patent allows the patient's own IgE antibodies to serve their natural function of binding to allergens without requiring external purification or activation steps. The IgE in sera is used directly in its native state, performing its physiological role of allergen recognition within the simplified detection system.
4Productivity
If IgE is directly bound to solid phase surface, then detection can be performed, but conformational changes occur that reduce binding affinity
Solution Approach 1:
The patent introduces an intermediary layer between the solid phase surface and IgE molecules. This intermediate layer prevents direct interaction between IgE and the surface, avoiding conformational changes that would reduce binding affinity. The intermediary maintains IgE in a native-like state while still enabling immobilization for detection.
Solution Approach 2:
The patent uses a thin film or flexible intermediary layer that allows IgE molecules to maintain their three-dimensional conformation while being immobilized. This flexible interface preserves the binding site structure of IgE, ensuring high binding affinity to allergens despite surface attachment.
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 rapid, safe, and cost-effective detection and identification of allergens and parasitic or fungal infections by preserving the native conformation of IgE and allergens, ensuring accurate interaction and binding, even in the presence of post-translational modifications.
Implementation Method 1
binding of the IgE to the solid phase surface via the IgE receptor FcεRlα-(His)6
Implementation Method 2
utilizing a recombinant manufacturing process and histidine tags to keep IgE at a sufficient distance from the surface
Implementation Method 3
allowing for the selective binding and detection of allergens through their conformational epitopes
Data Source
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AI summary
The invention relates to a method for the in-vitro detection of a parasitic or fungal infection using infection-specific IgE from blood plasma or blood serum, which are bound in native form to a solid phase surface and thus allow infection-specific allergens to be selectively bound and subsequently identified in native form, i.e. via the conformational epitopes.