Phage Display Allergen Epitope Mapping
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Solution Overview
Problem
Current methods for diagnosing food allergies, such as skin prick tests and component testing, are limited by low specificity and require large sample volumes, making it difficult to accurately identify allergenic epitopes and leading to unnecessary food avoidance and ineffective treatment strategies.
Innovation Solution
A programmable phage display-based method that analyzes anti-allergen IgE and IgG antibodies to 1,847 allergenic proteins using a library of overlapping 56 amino acid peptides, enabling high-throughput DNA sequencing and sensitive detection of specific allergenic motifs, thereby providing a more precise diagnosis of food allergies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If skin prick tests and component testing are used for food allergy diagnosis, then testing can be performed, but the specificity is low leading to false positive results
Solution Approach 1:
The patent segments allergenic proteins into overlapping 56 amino acid peptide tiles that comprehensively cover the entire protein sequence. This segmentation allows for precise mapping of IgE binding to specific epitopic regions rather than testing whole proteins, thereby improving both diagnosis specificity and epitope identification accuracy by localizing allergic responses to exact peptide sequences within allergens
Solution Approach 2:
The patent replaces traditional mechanical/physical testing methods (skin prick tests, whole protein component testing) with a molecular biology-based approach using phage display technology. This substitution enables direct detection and mapping of IgE antibody binding to specific peptide sequences through in vitro binding assays followed by DNA sequencing, achieving higher specificity by directly identifying the molecular basis of allergic responses rather than relying on physiological skin reactions or bulk protein binding
2Quantity of substance
If traditional component testing methods are used, then single allergens can be tested, but large sample volumes are required
Solution Approach 1:
The patent creates a universal phage display library that can simultaneously test against multiple allergens (1,847 different allergenic proteins) in a single reaction. The overlapping peptide tile design provides universal coverage across all allergen types, allowing the same library to detect IgE binding to any allergen without requiring separate tests for each allergen, thereby reducing sample volume requirements while increasing testing throughput
Solution Approach 2:
The patent merges the testing of multiple allergens into a single multiplexed assay. By combining overlapping peptide tiles from numerous different allergens into one comprehensive phage display library, the method allows simultaneous evaluation of IgE reactivity to many different allergens in one experiment, eliminating the need for separate component tests and significantly reducing the total sample volume required compared to traditional sequential testing approaches
3Measurement precision
If epitope mapping is performed to identify specific allergenic peptides, then diagnostic precision can be improved, but the process is costly and time-intensive
Solution Approach 1:
The patent replaces time-intensive traditional epitope mapping methods (such as sequential peptide array testing or structural biology approaches) with a high-throughput molecular biology method. By using phage display combined with next-generation DNA sequencing, the system can identify all IgE-binding epitopes in parallel through automated sequencing and bioinformatics analysis, achieving high epitope identification accuracy while reducing the time required from weeks to days
Solution Approach 2:
The patent changes the scale and capacity parameters of epitope mapping by using a massively parallel phage display system. The overlapping 56 amino acid peptide tile design ensures complete coverage of all possible epitopic regions, and the use of high-throughput sequencing allows simultaneous analysis of binding to thousands of different peptides, thereby achieving comprehensive and accurate epitope identification in a single experiment rather than through sequential testing
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
This approach allows for comprehensive analysis of allergenic peptides, improving the specificity of food allergy diagnosis, enabling personalized avoidance patterns and potential immunotherapy strategies, and reducing the cost and complexity of current methods.
Implementation Method 1
contacting a reaction sample comprising a display library with a biological sample comprising antibodies, wherein the display library comprises a plurality of peptides derived from a plurality of allergens
Data Source
AI summary
The present invention relates to the field of allergies. More specifically, the present invention provides compositions and methods useful for identifying anti-allergen antibodies in a patient sample using phage display. In one embodiment, a method for detecting the presence of an antibody against an allergen in subject includes the steps of (a) contacting a reaction sample comprising a display library with a biological sample comprising antibodies, wherein the display library includes a plurality of peptides derived from a plurality of allergens; and (b) detecting at least one antibody bound to at least one peptide expressed by the display library, thereby detecting an antibody against the at least one peptide in the biological sample.