Cleavable Luciferase Platform for Rapid Coronavirus Spike-ACE2 Detection
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Solution Overview
Problem
Current methods for detecting the interaction between the spike protein receptor binding domain of coronaviruses and human angiotensin-converting enzyme II (ACE2) are time-consuming, ranging from 3 to 5 days, posing a significant burden on research units and limiting accessibility due to the need for biosafety level 3 laboratory facilities.
Innovation Solution
A method involving the ligation of cleavable luciferase DNA fragments to the spike protein receptor binding domain and human ACE2 DNA, expressed in cells to detect luminescence signals, allowing for rapid detection of the interaction within 20 minutes using the NanoBiT technology.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If plaque assay is used to detect the interaction between spike protein receptor binding domain and ACE2, then measurement precision is improved, but loss of time increases significantly (3-5 days)
Solution Approach 1:
The patent replaces the traditional plaque assay method with a luminescence-based detection system using NanoBiT technology. This substitution transforms the mechanical/cell-based plaque formation process into an optical detection method, achieving rapid results within 20 minutes while maintaining detection accuracy through luminescence signal measurement.
Solution Approach 2:
The patent changes the detection parameter from plaque formation (visual/count-based) to luminescence intensity (optical signal). By using luciferase-based luminescence signals, the system achieves both speed and precision, resolving the contradiction between fast detection and accurate measurement.
2Measurement precision
If plaque assay is used for detection, then measurement precision is improved, but device complexity increases due to BSL-3 facility requirements
Solution Approach 1:
The patent employs a disposable luminescence detection system that does not require permanent BSL-3 facilities. The NanoBiT-based assay can be performed in lower biosafety levels, effectively replacing the need for complex, expensive BSL-3 infrastructure while maintaining detection precision through luminescence measurement.
3Measurement precision
If traditional detection methods are used, then measurement precision is maintained, but productivity decreases due to long detection time
Solution Approach 1:
The luminescence-based detection system allows for continuous processing of multiple samples without the time-consuming plaque formation step. Multiple samples can be detected in parallel using standard luminescence plate readers, significantly increasing productivity while maintaining measurement precision through consistent optical signal measurement.
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 enables quick and robust detection of the virus spike attachment to the ACE2 receptor, reducing detection time to 10 minutes, facilitating the screening of drug candidates for coronavirus infection and eliminating the need for high-level biosafety facilities.
Implementation Method 1
detecting the interaction between the spike protein receptor binding domain of coronavirus and the human angiotensin-converting enzyme II in the cell by detecting a luminescence signal
Data Source
AI summary
The present disclosure provides a method and a platform for enhancing detection activity of an interaction between a spike protein receptor binding domain of coronavirus from a specimen and a human angiotensin-converting enzyme II. The method and the platform of the present disclosure use a cleavable luciferase as a report test for the combination of the spike protein receptor binding domain of coronavirus (such as novel coronavirus) and angiotensin-converting enzyme II. Screening is carried out at the cellular level. The strength of the drug's influence on the interaction between the two molecules can be judged by the strength of the luminescence signal. The detection time can be completed within 20 minutes.