Recombinant Binding Protein for Dengue NS1 Detection
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Solution Overview
Problem
Current methods for detecting dengue virus, particularly using NS1 protein, face challenges such as instability in monoclonal antibody production, cross-reactions, and the need for complex equipment, limiting rapid and accurate diagnosis.
Innovation Solution
Development of an isolated binding protein with an antigen-binding domain that specifically binds to the NS1 protein, utilizing specific CDR sequences and an eukaryotic cell expression system for production, eliminating the need for mouse-derived hybridomas and enhancing stability and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If mouse-derived monoclonal antibodies are used for NS1 protein detection, then detection capability is achieved, but production stability is poor and batch-to-batch difference is great
Solution Approach 1:
The patent creates a recombinant binding protein that copies the antigen-binding function of mouse monoclonal antibodies against NS1 protein. By cloning the variable region genes (VH and VL) from the mouse antibody and expressing them as a recombinant protein in mammalian cells, the invention replicates the specific binding capability without requiring hybridoma production, thereby achieving stable and consistent batches
Solution Approach 2:
The patent replaces the biological mechanical system of hybridoma cell culture and antibody secretion with a recombinant protein expression system. Instead of relying on mouse cell fusion and in vivo antibody production, the invention uses in vitro transcription and translation in mammalian cells to produce the binding protein, eliminating the variability associated with hybridoma maintenance and antibody secretion
2Reliability
If mouse-derived monoclonal antibodies are used, then NS1 protein binding is achieved, but purification is difficult due to mouse autoantibodies
Solution Approach 1:
The patent extracts only the essential antigen-binding function from the mouse monoclonal antibody by cloning and expressing only the variable region genes (VH and VL) as a recombinant binding protein. This removes the constant regions and Fc portions that would otherwise bind to mouse autoantibodies and complicate purification, while retaining the specific antigen-binding capability
Solution Approach 2:
The recombinant binding protein acts as an intermediary that provides the specific antigen-binding function without the problematic components of mouse antibodies. By using a simplified protein structure expressed in a controlled system, it mediates the detection function while avoiding contamination from mouse autoantibodies that would interfere with purification
3Measurement precision
If conventional serological diagnosis methods are used, then detection is possible, but cross-reactions occur reducing accuracy
Solution Approach 1:
The patent applies local quality by focusing the binding specificity entirely on the variable regions (CDR1, CDR2, CDR3) of the antibody that are responsible for antigen recognition. By designing the recombinant binding protein to contain only these critical local regions with high specificity for NS1 protein, the invention achieves precise detection while minimizing cross-reactions with other antigens
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 binding protein achieves high sensitivity and specificity in detecting dengue virus, facilitating rapid and accurate diagnosis without the production instability and complexity associated with traditional methods.
Implementation Method 1
an antigen-binding domain that binds to NS1 protein
Data Source
AI summary
Provided is an isolated binding protein including an antigen-binding domain that binds to NS1 protein. The isolated binding protein includes specific heavy chain CDRs and light chain CDRs. The binding protein can specifically recognize and bind to NS1, and has relatively high sensitivity and specificity, thereby achieving the detection of dengue virus. Moreover, the binding protein is not required to be produced by inducing hybridoma cells in mouse abdominal cavity, and thus it is simple in production and has more stable antibody function.