NanoSensing POC Device TechnologyFAQs (Frequently Asked Questions) answeredQ1. The device is currently under the protection of a provisional patent. Are themethods or testing materials able to be patented as unique?Yes, they are. We developed a Point of Care (POC) device and a Point ofCare Test assay (POCT) for fast and accurate quantification of cardiacTroponin I (cTnI) directly in human blood plasma, or whole blood samples.Both POC and POCT make use of modified metallic nanoparticles,conjugated to anti-body Anti-cTnI. Localized Surface Plasmon Resonance(LSPR) phenomenon is an intrinsic feature of these modified metallicnanoparticles, which arises from the oscillation of free electrons in the metaland increases the sensitivity and intensity of the measured signal. In recentyears, biomedical research has received a large advancement, which greatlybenefit from the development of nanomaterials. In the case of metallicnanoparticles, they hold unique optical properties such as good biosafety,facile surface modification and the above mentioned LSPR. These featuresmake them quite suitable nanoscale platforms for biosensing and forqualitative/quantitative tests and diagnostics assays.Q2. Is the device universally applicable with other devices or is it a stand alone?- Our invention “Nanoplasmonic Point of Care Device for Troponin Iquantification in whole human blood or plasma” may be used in small clinics,in large hospitals and even for in home care patients assisted by credentialedprofessionals. Our POC device may be connected to other devices such asprinters, computers, laptops and others, by one of its USB ports.- Our invention “Spectroscopic Method for Troponine I detection andquantification by modified plasmonic nanomaterials” here named as POCT(due to the fact that it consists in a Point of Care Test) may be used inHospitals, Clinics, and in any other situation, such as in an ambulance, intransit accidents, by athletes during their sports activity (in recent years somesoccer players had died by heart attack during soccer matches. Besides this,many sports involve the release of large amounts of adrenaline. Excess ofadrenaline may induce cardiac injuries, including heart attacks), in renalpatients during hemodyalisis (in this case, the risk of heart failure is highlyincreased) and many other situations, in order to evaluate the risk of a heart
attack. It is well known that Troponin I starts circulating in the blood stream 6to 4 hours before a heart attack. The POCT assay simply makes use of a USBmini-spectrophotometer (commercially available); it includes specific softwarefor recording and displaying the data, a laptop (or any computer with USBports), the nanomaterial (which is already conjugated to Anti-Troponin I) anda small amount of blood (less than 0.5 mL). The result will be displayed in lessthan 4 minutes.Q3. Based on the existence of similar systems, is the company required toapply for US-FDA approval? If so, what is the status of this process?Yes, the device shall be submitted for US-FDA approval. This process hasjust started.Q4. Can this device move forward with a 510K application approval?We are not sure yet, we are inquiring the possibility with the regulatoryagencies.Q5. Outside of the primary application, have trials begun to identify other usesfor the device/technology?Yes, we’ve identified many others applications for the same technology. Inaddition to having the usage as fast (less than 4 minutes), the low bloodvolume (20 µL) cost-effective, and precise diagnostic of myocardial injuries(including early detection of heart attack risk), the above mentioned testplatforms may be adapted for the use in the diagnostic of a wide range ofpathologies, enabling early management and improved outcomes.These platforms may also be adapted for detecting DNA, RNA, biomarkers,bacteria and other pathogens in water, blood, urine and other samples. Someof the potential applications of our in vitro test platforms are listed below: Dengue fever (acute stage); Hepatites; Leishmaniosis; Chagas disease; Septicemia; Qualitative and quantitative detection of pathogens in healthcare settings (MRSA, UTI bacteria, etc.), in the food chain (E. coli,
Salmonella, and Listeria monocytogenes), and some pathogens and biomolecules used in bioterrorism (e.g., Bacilus anthracis (anthrax), poisonous protein ricin, etc.); HPV detection and typing; Qualitative and quantitative detection of specific compounds/chemical (DNA, RNA, antibiotics and other drugs) species in blood, saliva, urine, sweat and water.Adapting our test platforms for some of these applications may require veryspecific working conditions, including equipments and/or laboratorialinstallations with distinct biological security levels.Q6. Will the same device be able to be used for multiple applications bychanging the cuvette?Yes, the same device can be used for multiple applications by changing thecuvette (for POC device) or changing the metallic nanorods coverage (forPOCT assay).Q7. Does this test meet the requirement of ≤ 10% CV concentration at the99th percentile reference limit? Most commercial devices do not currently meetthis requirement and it has been noted and established as the level ofsignificance such technologies should strive to achieve, specifically to avoidfalse positive readings.Analytical evaluation was performed according to the protocols of the Clinicaland Laboratory Standards Institute, using control samples at 3 concentrationsof cTnI. In order to obtain the control samples, we used freshly collected (76patients) and stored (up to 20 days of storage; 46 patients) duplicated bloodsamples in which the cTnI concentration was previously measured by ELISAmethod and by using Abbott i-Stat (point of care device). The assaysperformed with our POC device presented a limit of detection of 0.006ng/mL and a CV of 10% at concentrations approaching 0.03 ng/mL. Thetotal CV did not exceed 6% at concentrations of 0.12, 0.48, and 11.08ng/mL. In order to evaluate the selectivity, we performed assays in which thePSS capped nanorods were conjugated to Anti-IgG and Anti-IgM (0.06ng/mL), using duplicate samples of freshly collected (76 patients) and stored(up to 20 days of storage; 46 patients).Most of the commercially available POC devices for cTnI detection make useof chemiluminescent properties of fluorescent organic compounds. These
compounds however present some inherent properties, which can beresponsible for false positive and even cause other experimentalinconveniences. These fluorescent organic probes present short fluorescencelife-time, high photobleaching rates and may interact nonspecifically withother compounds than the target compound or even with distinct parts of thesame target compound. Both the POCT assay and the POC device developedby NanoSensing make use of plasmonic properties which are present in somemetallic materials when in nanometric size range. Unlike fluorescence, LSPR(Localized Plasmon Resonance) is not a short life-time property and is highlyconnected to the electrical charge distribution of the sample. Even smallslightly distinct compounds do not present the same charge distribution. Thisway, whenever the metallic nanorod is excited by the correct light source, thesame optoelectronic signal will be generated, resulting so ever in the sameabsorption spectrum. This avoids false positive or false negatives as well asprovides more fast, specific and accurate responses.Q8. Is the future production and sale of the nanomaterial the driver of the deal?No, NanoSensing does not intend to make the products and/or nanomaterial.We enjoy developing technology and sell it to commercial partners.Q9. How easily could this solution/material be replicated?The device and the material can be easily replicated by the acquirer oftechnology. NanoSensing team can provide protocol documentation in whichevery step is clearly described.Q10. What is the preferred business model? Direct sale of thistechnology/device to a large manufacturer? Or would a licensing strategy bebetter?Direct sale of the technology is preferred.Q11. Would the scientists involved with the development be needed by thepotential acquirer? Or has enough testing been conducted to supply theacquirer with enough information to proceed directly with trials?The potential acquirer may be able to produce the material and the device,with or without our participation. This depends on the background of theacquirer team.
Q12. What testing must still be completed (FDA approval, Clinical trials, etc.)before this can be marketed as a complete product, ready for use in the medicalfield.FDA or equivalent approval.