Nanotech Covid19 Sensor

We aim to improve sensitivity, selectivity and accuracy of Lateral-Flow-Assay (LFA) Rapid-Test-Kits (RTKs) for Antigen Detection by applying a Hybrid of a colorimetric approach (typical of LFAs) with electrochemical detection using nanomaterials-enhanced electrodes [Figure 1]. Electrochemical signal clarity allows unambiguous indication of antigen presence, relegating the colorimetric reading as a secondary, backup signal. Colorimetric ambiguity is a known weakness in existing kit design, necessitating confirmatory RT-PCR. We propose a solution that is low-cost, scalable and does not require a clean room facility for electrode fabrication. If we succeed, all countries regardless of wealth and development status will be able to “test, test, test” without regard to current limitations of lab capacity and staffing, or having to deal with chronic shortages and unavailability of expensive/proprietary reagents for SARS-CoV-2 diagnosis. 

As of June 15, Malaysia recorded 8,494 cases and 121 deaths. Globally, 8,027,906 cases have been reported, with a total death of 436,273. The countries that have reported the highest number of cases are the United States, followed by Brazil and Russia. With rising SARS-CoV-2 infected cases, there is an unprecedented demand for the RT-PCR tests for COVID-19 diagnosis. However, most developing and under-developed countries were highly reliant on import supply chains or on export-restricted key inputs of RT-PCR tests for COVID-19. The situation has led to a huge gap between the demand and supply in these nations. Needless to say, the turnaround time for RT-PCR is about 24-48 hours and requires sophisticated lab and trained personnel.  Therefore, most countries including Malaysia have opted for an antigen test kit in a lateral-flow-assay platform for SARS-CoV-2 detection. However, lateral flow assay operates based on colorimetric principle and suffers with issues of sensitivity, selectivity and accuracy. The colorimetric result is difficult to be interpreted as they can result in faint bands at the test line, and does not quantify the virus in the tested clinical sample.  

We utilizes the “nanosensor dot” or the nSD in a Hybrid LFAs+Electrochemical RTK design (“Hybrid RTK”), to improve sensitivity, selectivity and accuracy, for SARS-CoV-2 detection vs typical antigen-based LFAs, working within the constraints of a developing country’s resources and infrastructure. The Hybrid RTK technology, with an integrated bioreader [Figure 1], incorporates three key elements:

(1) nSD transducer [Figure 2] provide 10x larger effective surface area for antibody attachment and, thus improving sensitivity by 10x.

(2) choice of antibodies obtained through computation modelling with high affinity to the spike proteins [Figure 3]

(3) immobilization of the specific antibodies on the nSD transducer for selectivity. For improved electrochemical transduction, we utilized electrochemical measurement techniques inspired from NASA SporeSat, where electrodes were immersed in agarose gel for calcium ions measurements from a single fern cell during 196 days in orbit (www.sporesat.org).

Our product aimed to serve all countries regardless of wealth and development status and without regard to current limitations of lab capacity and staffing or having to deal with chronic shortages and unavailability of expensive/proprietary reagents for SARS-CoV-2 diagnosis. We seek advice from medical professionals to understand the problem of current antigen test kits and their expectation for a sensitive, selective and accurate device that can provide results that are quicker than RT-PCR. Furthermore, we engage with medical front-liners in most hospitals and clinics to understand their limitations in developing countries that are often ill-equipped with the capacity for SARS-CoV-2 testing.

COVID-19 pandemic gravitated researchers to control SARS-CoV-2 spread. We aim to solve the issue of chronic shortage and unavailability of expensive/proprietary reagents for diagnosis. Herein, our product allows SARS-CoV-2 detection by applying a Hybrid of colorimetric approach (typical of LFAs) with electrochemical detection using nanomaterials-enhanced electrodes. The solution that we provide allows pervasive, timely, and accurate situational awareness for pandemic monitoring and response, which will be crucial as the world manages and mitigates the risk of subsequent major “waves” of COVID-19 until an effective vaccine is successfully trialled and made available at a global scale in the coming years.

To the user, the Hybrid RTK operates similarly to typical LFA, with several enhancements described below.

[1] Electrochemical Sensor Element: We use reduced graphene + conductive polymer nanocomposite, resulting in nSD fabricated using methods suitable for use in developing countries laboratories (screen printing and 3D printing), without requiring clean-room facilities. Since the fabrication of the transducer element was conducted in ambient conditions, we applied machine learning approaches to optimize the fabrication process. We showed that the nSD transducer can provide ten (10x) times larger effective surface area than regular electrodes, enabling the attachment of more antibodies per unit area, thus improving sensitivity by 10x. The nSD also demonstrated improved electron transfer kinetics, resulting in improved sensitivity.

To improve selectivity, we applied computational modeling to simulate the interaction between SARS-CoV-2 spike proteins and human receptors hACE, and two major interacting Spike protein residues with hydrogen bond occupancy of more than 50% were identified: Tyr449 and Tyr500, information which will be applied towards antibody-nSD integration.

[2] Silver Nanoparticle Conjugates: IgG antibodies that are conjugated to silver nanoparticles (AgNPs) will be placed at the conjugation pad. We use the nSD as a transducer to facilitate electron transfer from oxidized conjugated AgNPs at the test and control lines of the RTKs. The AgNPs allow for simultaneous colorimetric and electrochemical readings. The number of AgNPs corresponds to the strength of the electrochemical signal measured, improving sensor sensitivity. The electrochemical signal will be read by a bioreader, which can support multiple electrochemical electrodes. 

Our solution is dependent on the electrochemical sensor technology for the quantification of the SARS-CoV-2. The electrochemical detection of the virus is dependent on the nSD platform. The nSD platform provides ten (10x) times larger effective surface area than regular electrodes, enabling the attachment of more antibodies per unit area, thus improving sensitivity by 10x. The nSD also demonstrated improved electron transfer kinetics, resulting in improved sensitivity. We are coupling this technology with a colorimetric approach that is widely used in the lateral flow assay kits, in the hopes of providing results that are quicker than RT-PCR, without compromising on the sensitivity, selectivity and accuracy. 

nSD are deposited using electropolymerization techniques onto screen-printed carbon electrodes without losing signal sensitivity and stability, even under prolonged storage exceeding 30 days in aqueous media [1]. Also, drop-casting deposition methods were tested for noninvasive glucose sensing and were found to result in good sensitivity, selectivity, and detection limit towards glucose [2]. The nSD is versatile, able to be modified with other biomarkers for detection and quantification of various antigens, volatile compounds, and so forth, making nSD suitable to be modified for SARS-CoV-2 detection and quantification. For improved electrochemical transduction, we utilized electrochemical measurement techniques inspired from NASA SporeSat, where electrodes were immersed in agarose gel for calcium ions measurements from a single fern cell during 196 days in orbit [3], [4].

References:

[1] A. M. Benoudjit, M. M. Bader, W. W. A. Wan Salim. Study of electropolymerized PEDOT:PSS transducers for application as electrochemical sensors in aqueous media. Sen. Biosens. Res., 17, 18-24, 2018. https://doi-org.ezproxyberklee.flo.org/10.1016/j.sbsr.2018.01.001

[2] F. Abd-Wahab, H. F Abdul-Guthoos, W. W. A. Wan Salim. Solid-State rGO-PEDOT:PSS Transducing Material for Cost-Effective Enzymatic Sensing. Biosensors, MDPI, 9(1), 36, 2019. https://doi-org.ezproxyberklee.flo.org/10.3390/bios9010036

[3] J. Park, M. L. Salmi, W. W. A. Wan Salim, A. Rademacher, B. Wickizer, A. Schooley, J. Benton, A. Cantero, P. F. Argote, M. Ren, M. Zhang, D. M. Porterfield, A. J. Ricco, S. J. Roux and J. L. Rickus. An autonomous lab on a chip for space flight calibration of gravity-induced transcellular calcium polarization in single-cell fern spores. Lab Chip, 17, 1095-1103, 2017. https://doi-org.ezproxyberklee.flo.org/10.1039/C6LC01370H 

[4] https://www.nasa.gov/centers/ames/engineering/projects/sporesat.html, https://www.nasa.gov/ames/research/space-biosciences/sporesat-spacex-3-0

We opted for a non-linear approach in building our business theory of change.  

We are at the alpha-prototyping stage, and still working on enhancement of product features before we advance to product commercialization. Our next steps are to fabricate 500 test strips and 20 reader devices, using screen printing and 3D printing technique to rapidly scale production ahead of clinical trials. Deposition of nSD transducers and immobilization of antibodies will be conducted using simple electropolymerization techniques. The electrochemical signal will be read by a bioreader consisting of an open source programmable analog front end (AFE), the LMP91000, which can support multiple electrochemical electrodes and is programmed for amperometric/potentiometric measurement. We fabricate the bioreader using 3D printing and standard PCB fabrication.  Following the above, we will conduct field trials in collaboration with the Institute of Medical Research, under the Malaysian Ministry of Health. We are targeting a cost of USD 1 per test (at scale) with >90% accuracy in the field. The above-mentioned processes are expected to be completed within the first year.

Within the five years’ time, we expect to expand our business to market our product to other Southeast Asian countries such as Thailand, Indonesia, Vietnam, Philippines, where we have a huge customer base or to countries that are worst-hit by COVID-19 pandemic. We will also continuously perform R&D to introduce new features and to further miniaturize our product in the coming years. A continuous miniaturization & product feature enhancement process will allow us to stay relevant in the business, and not to be affected by other companies that introduce similar products. 

Our goal for next year is to complete beta-prototyping and proceed with pilot studies in Malaysia. The pilot studies include testing the feasibility of our product by conducting clinical trials in collaboration with the Institute of Medical Research, under the Malaysian Ministry of Health. By the end of next year, our hybrid RTK will be available at most hospitals/clinics in Malaysia.

Our goal for the next five years is to serve international markets, especially to Southeast Asian countries or other countries that are worst-hit by coronavirus pandemic. In this five years’ time, we will release our second version of hybrid RTK with enhanced sensitivity, selectivity and accuracy for virus detection and a further miniaturized version. 

Financial: We have a limited funding opportunity for research & development (R&D) of products, especially for those who want to proceed to the beta-prototyping stage. This process is mostly funded by the Government, while local industries are not engaging with universities for such R&D processes. Due to COVID-19 pandemic, the government is constrained and the applications for research funds are delayed. Malaysia's Prototype Research Grant Scheme Application Phase 2.0 for 2020 is expected to open only in late June 2020. 

Facilities: We are working with several research groups within International Islamic University Malaysia; however, we lack several facilities that are essential as we progress to the beta-prototyping stage.

Regulatory barriers: We need to validate the performance of our Hybrid RTK in the field. If our technology provides valuable outcomes in terms of virus quantification, we have to then pursue approval from the Ministry of Health for market authorization in Malaysia (MY). 

Financial: We have applied for a research grant from the Ministry of Science and Technology Malaysia and are currently waiting for the application decision. Meanwhile, we are also approaching angel investors for initial capital funding for us to proceed to the beta-prototyping stage.

Facilities: We are also planning to collaborate with Industrial partners to expedite our process of nSD characterization, as we apply for our current intended application.

Regulatory barriers: We will conduct field trials in collaboration with the Institute of Medical Research, under the Malaysian Ministry of Health. At the initial stage, we will be using the SARS-CoV-2 infected patient sample that has been stored in the laboratories. The virus quantity obtained from our Hybrid RTK will be compared with the results obtained from RT-PCR. We will then proceed with approval from the Ministry of Health for market authorization in Malaysia. We will also work with the Ministry to acquire exclusive rights in the supply of Hybrid RTKs to hospitals and clinicals throughout Malaysia.

CEO: Mr. Iqbal Shamsul

Chief Technologist: Dr. Amani Salim

Lead Scientists:

1. Dr. Mohd Hamzah bin Mohd Nasir
2. Dr. Izzat Fahimuddin Bin Mohamed Suffian
3. Dr. Azzmer Azzar bin Abdul Hamid
4. Dr. Mohd. Firdaus Bin Abd. Wahab

Junior Scientists:

1. Abdelmohsen Benoudjit
2. Piravin Raj Barthasarathy
3. Ihda Uswatun Shalihah Shohibuddin

[1] Mohd Iqbal Shamsul Kamar

Area of expertise: MIT-trained, former Wall Street energy trader and investment banker, now focusing on out-of-the-box and impactful market-driven solutions in healthcare, energy, deep tech, and social enterprise. Currently CEO and Founder of two start-ups in Malaysia: one working to deliver next-gen biosensors for medical and other applications, and the other mobilizes spare capacity in the healthcare sector to address supply-demand mismatches across public and private sectors and optimizes healthcare system utilization and health outcomes.

https://www.linkedin.com/in/iqbal-shamsul-40a601/

[2] Amani Salim, Ph.D

Area of expertise: sensor technologies with applications in agriculture, biology, environmental science, medicine and space biology.

www.amanisalim.com

[3] Mohd Hamzah bin Mohd Nasir, Ph.D

Area of expertise: Cellular biology, Neurobiology and Immunology in related to human infectious diseases, Malaria.

https://www.researchgate.net/profile/Mohd_Hamzah_Mohd_Nasirhttp://www.iium.edu.my/staff/show/5522

[4] Azzmer Azzar Abdul Hamid, PhD

Area of expertise: Structural Modeling & Rational Design of Environmental & Disease Proteins.

http://www.iium.edu.my/staff/show/7089https://scholar.google.com/citations?user=eHU3XpoAAAAJ&hl=en

[5] Izzat Fahimuddin Mohamed Suffian, PhD

Area of expertise: Nanomedicine, Drug Delivery, Protein Engineering, Genetic Engineering, Oncology.

http://www.iium.edu.my/staff/show/7836https://scholar.google.com/citations?user=ozwI7aUAAAAJ&hl=en

[6] Dr. Firdaus Abdul Wahab, PhD

http://www.iium.edu.my/staff/show/5236

[7] Abdel Mohsen Benoudjit

https://scholar.google.com/citations?user=PmApF3oAAAAJ&hl=en

[8] Piravin Raj Barthasarathy

www.linkedin.com/in/rajpiravin

www.rajpiravin.com

[9] Ihda Uswatun Shalihah Shohibuddin

www.linkedin.com/in/ihda-uswatun-shalihah-shohibuddin-31670413a/

www.researchgate.net/profile/Ihda_Uswatun_Shalihah_Shohibuddin

1. Department of Biotechnology Engineering, Faculty of Engineering, International Islamic University Malaysia.
2. Faculty of Pharmacy, International Islamic University Malaysia
3. Faculty of Science, International Islamic University Malaysia. 

We are a startup company that began as a means to undertake the technology commercialisation mission based around the technology portfolio developed by our lead scientist and technologist, Dr Amani Salim, primarily around nanotechnology-powered advanced materials and devices for biosensing.

We have been primarily focused on commercialising a saliva-based glucose sensor as well as environmental monitoring use cases until the COVID19 pandemic overtook the world. Our technology allows us to deliver the following key USPs:

• superior sensitivity (increase true positive rate) and selectivity (increase true negative rates)
• considerably lower detection limits vs existing methods, to broaden the detection window
• does not require specialised infrastructure / facilities to be used and/or operated
• low cost to manufacture and use, without the use of clean rooms or advanced facilities, with,
• target price is USD 1 per strip (on par with glucose strips) and low-cost readers (~USD 10 each) when manufactured at scale,
• minimise "re-inventing the wheel", i.e. use existing off-the-shelf methods, materials, techniques wherever possible.

The business models include:

• IP licensing: develop a patent portfolio and validated, market-ready reference designs and manufacturing specifications/schematics to be monetized with big pharma / medtech who will undertake regulatory approvals for consumer device approvals + manufacturing + sales / marketing + distribution
• OEM: design, manufacture and R&D different nSD use case implementations and developing, manufacturing them for integration into existing / new devices for consumer, point-of-care, security, defence, IoT, etc. 
• Full-value chain: full vertical integration to pursue direct consumer markets in healthcare, defence, IoT, etc

Combination of government early stage / seed technology development funding, followed by investor and venture capital until commercialisation / monetisation event(s)

• Mentorship
• Recognition by MIT
• Platform to connect with potential investors
• Potential collaboration with MIT & Harvard network of researchers

WE believe our technology opens new avenue and approaches for generating abundance and impact in healthcare, climate change, etc. We therefore wish to engage with partners who can connect us with other visionaries, change makers, and investors, as well potential  commercial and manufacturing partners, IP management partners who are already making waves in domains as we are looking to make an impact in. 

• MIT.nano
• Gates Foundation
• DARPA
• BARDA  

To fund prototype development and clinical trials.

Dr. Amani Salim is an advocate for women in science, especially focuses on empowering women in developing countries. The prize will be used for hiring more women scientists to lead technology-driven projects in nanoSkunkworkX and to develop mentorships programs for girls by partnering with local education experts.

Our sensor technology is targeted to be low-cost, high impact, scalable, versatile, and extendable while being able to be retrofitted into existing product packages. The initial focus is on a key SDG goal on health and wellbeing, which has been a core focus in our development efforts thus far. 

Even before we started on the goal of completely upending the COVID19 diagnostics game, we started by approaching the problem of low-cost, non-invasive diabetes monitoring using saliva. 

Additional applications we are exploring are focused on new frontiers in medical science that leverages the capabilities of our sensor to provide early detection of neural tube defects in the womb, for example. 

For the future, can we start to detect changes in blood biomarkers hours before a heart attack? What new detect breast or ovarian cancer markers can be detected using our tech, 100x more sensitive than conventional ones? 

So we look forward to engaging the public, industry, as well as the healthcare community to expand our scope and develop partnerships into the most high-impact use cases that we are likely not even considering at this point. 

Our sensor technology is targeted to be low-cost, high impact, scalable, versatile, and extendable while being able to be retrofitted into existing product packages. The initial focus in on a key SDG goal on health and wellbeing, which has been a core focus in our development efforts thus far. 

Even before we started on the goal of completely upending the COVID19 diagnostics game, we started by approaching the problem of low-cost, non-invasive diabetes monitoring using saliva. 

Additional applications we are exploring are focused on new frontiers in medical science that leverages on the capabilities of our sensor to provide early detection of neural tube defects in the womb, for example. 

For the future, can we start to detect changes in blood biomarkers hours before a heart attack? What new cancer markers can be detected using our tech, 100x more sensitive than conventional ones? 

So we look forward to engaging the public, industry, as well as the healthcare community to expand our scope and develop partnerships into the most high-impact use cases that we are likely not even considering at this point. 

We have targeted our technology solutions to be impactful to the COVID19 diagnostics issue, hitting all the major points on cost-effectiveness, scalability, versatility, and extendability while being able to be retrofitted into existing product packages. We are concerned about the financial sustainability of our effort, and wish to apply the tech into other problems outside the immediate pandemic. 

Thus, we are pursuing other applications to our tech can be profit-focused and redirecting generated revenue into further R&D and (if possible) other efforts related to leveraging the core sensor technology for AI, IoT, and other platforms that enhance the value proposition for our tech and ancillary technologies / applications. 

For example, even before we pursued the COVID19 diagnostics use case, we were pursuing funding to develop the technology for non-invasive diabetes monitoring using saliva, and we have been approached by end users to develop use cases in security/defence, environmental monitoring, etc. 

We are keen to seek partnerships to flesh out these use cases in terms of funding, but also in terms of accessing domain expertise and industry/sector access to both define and refine the target applications and accessing end-users and high-impact applications in those sectors.