Recombinant NS1 Binding Protein for Dengue Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for diagnosing dengue fever, particularly those involving the NS1 protein, face challenges such as unstable production of mouse-derived monoclonal antibodies and interference from cross-reactions in conventional serological tests.
Innovation Solution
Development of an isolated binding protein with an antigen-binding domain that specifically binds to the NS1 protein, utilizing a newly obtained anti-dengue virus NS1 7D9 monoclonal antibody, and establishing an eukaryotic cell expression system for production and purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If mouse-derived monoclonal antibodies are used for NS1 protein detection, then the detection method can be established, but the production is unstable and shows large batch-to-batch differences
Solution Approach 1:
The patent creates a simplified copy of the antibody binding function using a recombinant binding protein produced in eukaryotic cells. Instead of using complex mouse hybridoma systems, the invention copies the essential antigen-binding capability into a recombinant protein format that can be produced more consistently through cell culture and purification processes.
Solution Approach 2:
The patent changes the production parameters from animal-based hybridoma culture to recombinant expression in eukaryotic cells. This parameter change includes shifting from in vivo mouse abdominal cavity production to in vitro cell culture systems, enabling better control over production conditions and reducing batch-to-batch variability.
2Measurement precision
If conventional serological tests are used for dengue diagnosis, then the detection can be performed, but cross-reactions interfere with the results
Solution Approach 1:
The patent extracts the specific antigen-binding capability from the complex mouse monoclonal antibody system and isolates it into a recombinant binding protein. This extraction allows the essential detection function to be separated from the sources of cross-reaction interference present in conventional serological tests.
Solution Approach 2:
The recombinant binding protein serves as an intermediary between the NS1 antigen and the detection system. This intermediary provides specific binding to the target antigen while avoiding the cross-reaction problems associated with conventional mouse-derived antibodies and serological test systems.
3Ease of manufacture
If mouse monoclonal antibodies are produced in abdominal cavity, then the antibodies can be obtained, but purification is difficult due to mouse autoantibodies
Solution Approach 1:
The patent copies the antibody binding function into a recombinant protein system that does not produce autoantibody contamination. The recombinant binding protein is produced in eukaryotic cells through controlled expression, eliminating the source of mouse autoantibodies that complicate purification.
Solution Approach 2:
The patent converts the limitation of not being able to use mouse abdominal cavity production into a benefit by using recombinant expression systems. This alternative approach eliminates the harmful autoantibody contamination while maintaining the essential antigen-binding function.
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 fever virus, with improved stability and reduced batch-to-batch variations compared to traditional mouse-derived antibodies.
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 binding to an NS1 protein, and including specific heavy chain CDR and light chain CDR. The binding protein can specifically identify and bind to NS1, and has relatively high sensitivity and specificity, so as to detect dengue vims. Moreover, the binding protein does not need to be produced by injecting hybridoma cells into mouse peritoneal cavity, while simplifying production, thus stabilizing antibody functionality.