Biotinylated HCV Antigen Tetramers for B Cell Detection
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Solution Overview
Problem
Current methods lack the ability to directly examine and characterize Hepatitis C virus (HCV)-specific B cells, hindering the understanding of the humoral immune response during acute HCV infection and the evaluation of vaccine efficacy.
Innovation Solution
Development of biotinylated monomers and tetramers based on HCV antigens, specifically the E2 glycoprotein, to identify and isolate antigen-specific B cells, allowing for deeper characterization of the B cell response.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to study HCV infection, then general immune response can be measured, but specific B cell characterization is not possible
Solution Approach 1:
The patent uses biotinylated HCV antigen monomers as intermediaries that bind to HCV-specific B cells. These monomers serve as mediators between the target B cells and the detection system (streptavidin-fluorochrome conjugates), enabling specific identification and characterization of HCV-specific B cells that was not possible with conventional methods
Solution Approach 2:
The patent changes the physical-chemical parameters of the antigen by biotinylating it, which adds a biotin moiety to the HCV antigen monomer. This parameter change enables the antigen to be detected by streptavidin-conjugated fluorochromes, transforming it into a detectable tetramer complex that can specifically identify HCV-specific B cells
2Measurement precision
If HCV antigen is used directly for detection, then simplicity is maintained, but sensitivity and specificity are insufficient
Solution Approach 1:
The patent merges four HCV antigen monomers into a tetramer structure through biotin-streptavidin interactions. This combining of multiple monomers into a tetramer complex enhances the detection precision and sensitivity while managing the complexity through a systematic assembly process
Solution Approach 2:
The patent creates a composite detection system consisting of biotinylated HCV antigen monomers, streptavidin, and fluorochrome conjugates. This composite structure combines multiple components with different functions (antigen recognition, binding, and fluorescence detection) to achieve superior 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 the specific and sensitive detection of HCV-specific B cells, providing insights into the humoral immune response and vaccine efficacy, and facilitating the isolation and cloning of antibodies.
Implementation Method 1
contacting the sample with the tetramer under conditions which allow the tetramer to bind to any antigen specific B cells in the sample
Data Source
AI summary
The present disclosure relates to constructs useful in expressing biotinylated monomers and tetramers produced from these monomers. The present disclosure also relates to methods for production and use of these tetramers in identifying and isolating antigen specific B cells and cloning antibodies thereto.