Braveheart is pleased to provide an update concerning the development of a COVID-19 test with the University of Sheffield.
Aptamer successfully synthesised and delivered
The required affinity macromolecule (an aptamer which binds to the SPIKE glycoprotein on the surface of the virus) has now been successfully synthesised and supplied toParaytec.
The test involves the deployment of a DNA aptamer – a short, single-stranded DNA molecule that adopts a shape that is complementary to that of the virus surface protein. By coating a test surface with millions of copies of the aptamer molecule, Paraytec’s CAPTURE module can trap Covid-19 virus particles from a swab sample, as they flow over the test surface. Trapped viruses are then instantaneously coated with the SIGNAL GENERATION module, a synthetic chimeric protein, that both recognises the viral surface as well as being highly fluorescent. The resultant fluorescent signal is measured by the DETECTION module using a version of Paraytec’s patented Actipix area imaging technology configured for fluorescence detection.
In order to avoid using live Coronavirus in the lab during proof of concept, an artificial version of the virus is used, the surface of which mimics the Covid-19 virus, by displaying the precise protein (SPIKE) on it that the virus uses to gain access to human cells. The CAPTURE module comprises a fluidics system in which a specialised surface is coated with the COVID-19 DNA aptamer. Virus particles are trapped on the surface by the DNA aptamer. In order to detect trapped virus, Paraytec’s SIGNAL GENERATION module uses a synthetic bi-functional protein, that both recognises the virus as well as emitting light. This enables trapped viruses to be visualised by a version of Paraytec’s ActiPix system that detects UV and visible light as well as fluorescence. The CAPTURE/SIGNAL GENERATION modules, in the presence of the virus SPIKE coat protein, generates a specific light signature, which may be detected by ActiPix.
The development project involves a number of Work Packages (WP), summarised here:
WP1. Construction of a viral mimic
In order to avoid using live Coronavirus in the lab during proof of concept, an artificial version of the virus is used, the surface of which mimics the Covid-19 virus, by displaying the precise protein (SPIKE) on it that the virus uses to gain access to human cells.
WP2. Construction of CAPTURE module
The CAPTURE module comprises a fluidics system in which a specialised surface is coated with the COVID-19 DNA aptamer. Paraytec intends to optimise the means by which virus particles are trapped on the surface by the DNA aptamer.
WP3 Construction of SIGNAL GENERATION module
In order to detect trapped virus, a synthetic bi-functional protein is used, that both recognises the virus as well as emitting light. This enables trapped viruses to be visualised by a version of Paraytec’s ActiPix system that detects UV and visible light as well as fluorescence.
WP4 Proof-of-concept testing of viral mimic/CAPTURE/SIGNAL GENERATION system
Perform flow cytometry analyses on a range of viral mimic densities to test the CAPTURE/SIGNAL GENERATION module.
WP5 Configuration of ActiPix for fluorescence signal DETECTION
Adapt the currently available ActiPix system with appropriate fluorescence excitation and emission filters. Build system with flow-through cuvette and test designs for a disposable cartridge.
WP6 Software refinement for data collection and analysis
Adapt current software for data collection, using fluorescence. Test to determine detection limit, using viral mimic at a range of levels in presence of unbound fluorescent protein.
WP7 System integration and sample handling.
Integration of viral mimic/CAPTURE/SIGNAL GENERATION system from WP4, detection system from WP5 and software from WP6 in order to create a single instrument and prototype disposable cartridge, fit for use in initial clinical study.
WP8 Validation of integrated system using (a) viral mimic and (b) UV-irradiated COVID-19 viral particles
Determination of lower sensitivity limits and dynamic range of detection for both viral mimic and irradiated COVID-19 containing samples.
WP9 Risk Management/Useability/Regulatory compliance
This work package will continue in parallel to ensure the necessary protocols and approvals are in place to undertake the first clinical study.
A further update will be provided after approximately 1 month, at which point Paraytec expects to have achieved the following:
I. Production of SPIKE protein in cell factories (WP1).
II. Modification of test surface with aptamer (WP2).
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